Search results for “assay

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80 articles

Comparative Analysis of Five Commercial RT-PCR Diagnostic Assay for Detection of Covid-19

Sep 2023 DOI 10.14302/issn.2692-1537.ijcv-23-4660
Amel Jamehdar SaeidCorresponding author

SARS-CoV-2 real-time reverse-transcription PCR (rRT-PCR) is the most effective testing system available to combat COVID-19, given the absence of any specific treatment or vaccine. Moreover, numerous SARS-CoV-2 rRT-PCR kits have been approved under emergency-use-authorization (EUA) worldwide. In this article, we present a comparison of important performance features among five commercial RT-PCR assays. A total of consecutive nasopharyngeal (NPS) samples and oropharyngeal (OP) swabs were collected from 50 COVID-19 patients to analyze sensitivity and specificity. The results showed variations in sensitivity among all the RT-PCR kits examined. The Pishtaz teb assays demonstrated the highest positive percent agreement (PPA) of 95.2% (40/42), followed by Covitech (90.5% - 38/42), DaAn Gene (83.3% - 35/42), Sansure (66.66% - 28/42), and Power check of SARS- CoV-2 panel (64.3% - 27/42). Conversely, all five molecular assays demonstrated a negative percent agreement (NPA) of 100% (8/8). These findings provide a technical baseline for assessing the performance of five distinct commercial PCR assays for detecting SARS-CoV-2. They could prove practical and useful for laboratories seeking to purchase any of these assays for further clinical validation. Highlights ·Compared five COVID-19 RT-PCR kits approved and available by Iran Ministry of Health. ·Pishtaz teb's kit identified the highest number of positive clinical samples.

The Evolution of the Enzyme Immunoassay/Enzyme-Linked Immunosorbent Assay

Aug 2021 DOI 10.14302/issn.2326-0793.jpgr-21-3917
Tarassishin LeonidCorresponding author Department of Pathology, Albert Einstein College of Medicine, United States.

50 years ago the Enzyme Immunoassay Enzyme-Linked Immunosorbent Assay, mostly known as ELISA was developed. This is a powerful but simple method that is very widely used in the diagnostic practice, as well as in biomedical research. During this time a number of ELISA modification were developed that significantly increased its properties, especially the senstivity, such as avidin-biotin assay, immuno-PCR, nano-ELISA and finally, the digital ELISA. This short review describes the principles of ELISA and the evolution from a conventional assay to the modern ultra-sensitive method. Most of the immunological methods have two components: antigen and antibody. The high specificity of their interaction gives a possibility to detect one of them if other one is included in the reaction as a specific partner. The simplest method for antigen detection in the presence of the antibody is immune diffusion (radial immune diffusion in that case), which practically the formation of precipitate of the “antigen-antibody” complex, when the target antigen diffuses from well into agarose containing the specific antibody. Unfortunately, this assay, as well as other traditional methods, like hemagglutination or complement fixation, have a low sensitivity and are unwieldy.

The Feasibility of Enzyme Immunoassay Tests in the Absence of a Conventional Source of Electricity

Oct 2020 DOI 10.14302/issn.2576-6694.jbbs-20-3517
S.U. DOSSOU CamilleCorresponding author Laboratoire de Recherche en Biologie Appliquée, Ecole Polytechnique d’Abomey-Calavi (EPAC), University of Abomey-Calavi, 01 BP 2009 Cotonou, Benin.

Background of the Study African  countries are facing frequent blackout. Thus in sub Saharan region, due to frequent power cut, the laboratory professionals find sometimes difficulty to carry out earlier diverse diagnostic tests. Objective The  aim of this work is to evaluate the feasibility of enzyme immunoassay tests in the absence of a conventional source of electricity. Methods We  developed a battery-powered experimental device, which was then applied to diagnose measles. The samples included 45 sera randomly selected from non-haemolysed serum samples received and stored at the National Public Health Laboratory of Benin. The experimental device is composed of two devices (Devices 1 and 2). The Device 1 provided an average temperature of 34.47 °C, 20 min after starting. With Device 2 an average temperature of 20.32 °C is obtained 15 min after starting. Results With  the experimental device the same rate of measles antibody-positive sera (44.68%) was obtained as recorded from the test using the standard equipment of laboratory. The experimental device detected 18 negative and 8 intermediate results against respectively 19 and 7 by the standard equipment. The analysis of the results of both equipments shows a concordance rate of 93.33% with a kappa reproducibility coefficient of 0.89. Conclusion The  device conceived in our study is a simply equipment allowing the realization of the enzyme immunoassay tests, in this case the ELISA anti-measles test. The rate of concordance obtained shows that this device can be used with commercial kits and at temperatures close to those recommended by the manufacturer without altering the results.

Use of Microfluidic Assays to Develop Reliable and Economic Nucleic Acid Application Technologies, Employing MicroRNAs for the Diagnostic Screening of Colon Cancer in Human Stool in Low-Resource Settings

Oct 2020 DOI 10.14302/issn.2379-7835.ijn-20-3418
E. Ahmed FaridCorresponding author GEM Tox Labs, Institute for Research in Biotechnology, 2905 South Memorial Drive, Greenville, NC 27834, USA

Isolation methods that employ readily-available inexpensive supplies on the open market, which are reliable, as well as economical, such as nucleic acid amplification techniques (NAAT) based on microfluidic technology in low-resource research settings (LRRS) that meets the ASSURED guidelines are essential to develop a noninvasive diagnostic colon cancer screen in stool using micro(mi)RNA molecules. A combination of a microfluidic-based MiRNA stool test with a reliable rolling circle amplification/detection method applied to the quantification of miRNA molecules, result in an affordable sensitive and specific isothermal method for the noninvasive quantitative detection of miRNAs in LRRS. Scientists and engineers have become interested in miRNAs, and they have intensified their efforts to apply emerging simple detection tools to the important bioanalytical challenge of quantifying these small 18-26 nt long molecules. Some of the proposed approaches incorporate novel material, such as simple centrifuges and methods based on microfluidic technology, while others utilize the interesting biological properties of these molecules, such as forming branched RCA structures, allowing for the detection of these biomarker molecules at an attomolar "aM" concentration level, using low cost extraction and isothermal amplification methods in LRRS. We have been interested in studying colorectal cancer (CRC) because it is the 3rd most common malignancy worldwide, and stool can be obtained noninvasively from the patients. We have focused in this research on colon cancer (CC) because it is more common in the USA than rectal cancer (RC). The innovation of our approach lies in the exploratory use of an affordable, quantitative miRNA profiling in noninvasive stool samples in LRRS, whose extracted fragile total RNA is stabilized shortly after excretion from stool by commercially available kits, so it does not ever fragment, followed by quantitative standardized analytical tests that are neither labor intensive, nor require expensive instrumentation, in order to develop apanel of novel miRNA genes for the noninvasive diagnostic screening of early left and right sporadic colon cancers, more economically, and with higher sensitivity and specificity than any other colon cancer screening test currently available on the market. To show the clinical sensitivity and specificity of the proposed quantitative miRNA test using simple methodologies in LRRS,the miRNA results are to be correlated with FOBT, colonoscopy, and pathology data. Standardization establishes test’s performance criteria (sample selection, optimal sample running conditions, preservation and storage), in order to ensure that the assay will perform the same way in any laboratory, by any trained personnel, anywhere in low-resource laboratory settings worldwide.

Use of Microfluidic Assays to Develop Reliable and Economic Nucleic Acid Application Technologies, Employing MicroRNAs for the Diagnostic Screening of Colon Cancer in Human Stool in Low-Resource Settings

Jun 2020 DOI 10.14302/issn.2379-7835.ijn-19-3123
E. Ahmed FaridCorresponding author GEM Tox Labs, Institute for Research in Biotechnology, 2905 South Memorial Drive, Greenville, NC 27834, USA

Isolation methods that employ readily-available inexpensive supplies on the open market, which are reliable, as well as economical, such as nucleic acid amplification techniques (NAAT) based on microfluidic technology in low-resource research settings (LRRS) that meets the ASSURED guidelines are essential to develop a noninvasive diagnostic colon cancer screen in stool using micro(mi)RNA molecules. A combination of a microfluidic-based MiRNA stool test with a reliable rolling circle amplification/detection method applied to the quantification of miRNA molecules, result in an affordable sensitive and specific isothermal method for the noninvasive quantitative detection of miRNAs in LRRS. Scientists and engineers have become interested in miRNAs, and they have intensified their efforts to apply emerging simple detection tools to the important bioanalytical challenge of quantifying these small 18-26 nt long molecules. Some of the proposed approaches incorporate novel material, such as simple centrifuges and methods based on microfluidic technology, while others utilize the interesting biological properties of these molecules, such as forming branched RCA structures, allowing for the detection of these biomarker molecules at an attomolar "aM" concentration level, using low cost extraction and isothermal amplification methods in LRRS. We have been interested in studying colorectal cancer (CRC) because it is the 3rd most common malignancy worldwide, and stool can be obtained noninvasively from the patients. We have focused in this research on colon cancer (CC) because it is more common in the USA than rectal cancer (RC). The innovation of our approach lies in the exploratory use of an affordable, quantitative miRNA profiling in noninvasive stool samples in LRRS, whose extracted fragile total RNA is stabilized shortly after excretion from stool by commercially available kits, so it does not ever fragment, followed by quantitative standardized analytical tests that are neither labor intensive, nor require expensive instrumentation, in order to develop apanel of novel miRNA genes for the noninvasive diagnostic screening of early left and right sporadic colon cancers, more economically, and with higher sensitivity and specificity than any other colon cancer screening test currently available on the market. To show the clinical sensitivity and specificity of the proposed quantitative miRNA test using simple methodologies in LRRS,the miRNA results are to be correlated with FOBT, colonoscopy, and pathology data. Standardization establishes test’s performance criteria (sample selection, optimal sample running conditions, preservation and storage), in order to ensure that the assay will perform the same way in any laboratory, by any trained personnel, anywhere in low-resource laboratory settings worldwide.  

The Current Immunoassays and Emerging Immunogenomic Approaches for Immunomonitoring Cancer and Infectious Diseases

Jan 2019 DOI 10.14302/issn.2572-3030.jcgb-18-2527
Zhang XiCorresponding author Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, United States

As remarkable advances have been made in immunotherapies, the overall goal of immunotherapy has become the selection of patients and evaluating the benefits of treatment. One of the major obstacles to develop immunotherapies is the lack of effective immune monitoring. Monitoring of key changes in the immune system during immunotherapy (immunomonitoring) provides important insights into efficacy as well as the immune mechanisms of response at the molecular and cellular levels. Immunomonitoring techniques include traditional immunoassays that use specific antibodies to recognize the analytes of interest, new high-throughput immunoassays that target immune cells and nucleic acids, and less classical immunogenomic approaches that rely on genome-wide profiling and computational analysis on various types of clinical samples. Substantial progress has been made in the application of immunomonitoring strategies to pre-clinical and clinical studies, especially for patients with cancer and infectious diseases. Current and emerging immunoassays performed in clinical practice will be examined herein, and immunogenomic approaches that complement these techniques will be highlighted and compared with traditional methods. Finally, we will discuss several new computational methods for analyzing gene signatures for immunomonitoring, including gene expression data profiling by microarray, the nCounter technique, regular RNA-seq, and single-cell RNA-seq. Novel immunomonitoring techniques, especially immunogenomic approaches, will continue to be developed to facilitate assessment of immunotherapeutic response and predict patient outcomes in cancer and infectious disease.  

The Development and Evaluation of A Multiplex Real-Time PCR Assay for the Detection of ESBL Genes in Urinary Tract Infections

Aug 2018 DOI 10.14302/issn.2690-4721.ijcm-18-2217
Samaras ShivanthiCorresponding author Molecular Microbiology, School of Allied Health Sciences, Faculty of Health & Life Sciences, Hawthorn Building, The Gateway, De Montfort University, Leicester, LE1 9BH United Kingdom

Background Overuse of beta-lactam antibiotics has lead to selection for extended-spectrum β-lactamase (ESBL) producing Enterobacteriaceae, a major cause of antibiotic resistant urinary tract infections (UTIs). Standard detection methods are time-consuming, with disputed accuracy. This study describes a novel real-time PCR method to detect CTX-M, SHV, OXA and TEM. Methods 179 Enterobacteriaceae isolates from UTIs were collected from the Leicester Royal Infirmary, UK. A multiplex Plexor®-based real-time PCR assay detected ESBLs using their specific amplicon melting temperature, during each cycle, removing the need for a melt-curve analysis. Validation was achieved by end-point PCR and disk diffusion. Results The method was able to produce rapid and accurate results, achieving a sensitivity and specificity of 94.9% and 72% respectively, and the assay can differentiate between the different ESBL genes, with ease. Conclusions With further investigation, a Plexor®-based assay could form the basis of a high-throughput kit that health services could use to detect ESBLs or other antibiotic resistance genes.

Polysaccharide Transglycosylases: A Survey of Assay Methods

May 2018 DOI 10.14302/issn.2832-5311.jpcd-18-2077
Farkaš VladimírCorresponding author Institute of Chemistry, Centre for Glycomics, Slovak Academy of Sciences, 84538 Bratislava, Slovakia

Polysaccharide transglycosylases (PTGs) are a unique group of glycoside hydrolases playing important roles in the formation and modification of plant and fungal cell walls. Their action involves cutting the molecule of the polysaccharide substrate at the glycosidic bond, followed by transfer of the newly formed reducing-end fragment to the non-reducing end of another polysaccharide molecule, with the formation of a new glycosidic bond. As there is no net increase in the number of reducing ends in the system, conventional reductometric methods used to assess the activity of glycoside hydrolases are ineffective. Since the PTGs participate in vital processes, such as the elaboration of cell walls in plants and fungi, and are not present in animal cells, they are considered as possible targets for future specific fungicides and herbicides. Biochemical studies of PTGs, as well as the search for their inhibitors, require the availability of convenient and efficient methods for their assay. In this review we briefly describe the principles of methods used to detect and to determine the activity of this important group of enzymes.

Calcium Transient Assays for Compound Screening with Human iPSC-derived Cardiomyocytes: Evaluating New Tools

Jan 2017 DOI 10.14302/issn.2574-4372.jesr-16-1395
Wakatsuki TetsuroCorresponding author InvivoSciences, Inc., Madison, WI

Calcium (Ca2+) plays a central role in regulating many biological processes in the cell from muscle contraction to neurotransmitter release. The need for reliable fluorescent calcium indicator dyes is of vast importance for studying many aspects of cell biology as well as screening compounds using phenotypic high throughput assays. We have assessed two of the latest generation of calcium indicator dyes, FLIPR Calcium 6 and Cal-520 AM for studying calcium transients (CaTs) in induced pluripotent stem cell (iPSC) -derived human cardiomyocytes. FLIPR Calcium 6 and Cal-520 dyes both displayed robust CaTs with a high signal-to-noise ratio (SNR) and were non-toxic to the cells. The analysis showed that CaT amplitudes were stable between measurements, but CaT duration was more variable and tended to increase between reads. Two methods were compared for drug-screening hit-selection; difference in average (unstandardized) and standardized difference. The unstandardized difference was better for assessing CaT amplitude, whereas standardized difference was equal to or better for assessing CaT duration. In summary, FLIPR Calcium 6 and Cal-520 are suitable dyes for drug-screening using iPSC-derived human cardiomyocytes.

Correlation of Cryptococcal Antigen Assay with C-reactive Protein as Serum and Urine Biomarker in Cryptococcal Meningitis: Experience in a Tertiary Hospital

Jul 2013 DOI 10.14302/issn.2324-7339.jcrhap-12-150
Kashyap BineetaCorresponding author Department of Microbiology

Aims and Objectives: The incidence of cryptococcal meningitis caused by Cryptococcus neoformans has risen markedly over the past 20 years as a result of theHIV epidemic and increasing use of immunosuppressive therapies. The objectives of this study were to isolate and identify Cryptococcus neoformans from clinically suspected cases of fungal meningitis by conventional techniques and evaluate the role of C-reactive protein (CRP) as a serum or urine biomarker for the diagnosis and monitoring of patients with cryptococcal meningitis. Materials and Methods: Direct microscopic examination of the CSF samples from clinically suspected cases of fungal meningitis was done by India Ink staining for the capsule demonstration and isolation of the Cryptococcus neoformans was done by inoculation of the sample on Sabourauds dextrose agar. Latex agglutination test for the presence of cryptococcal antigen was done on sera, CSF and urine samples. C Reactive Protein levels were estimated in sera and urine. Result: Cryptococcal meningitis was diagnosed in 12 cases by culture and/or India Ink staining and/or latex agglutination assay for antigen detection in CSF. Only 8 (66.67%) and 1 (8.33 %) out of 12 samples were positive for cryptococcal antigen in sera and urine respectively. Whereas all the 12 patients were positive in the sera for CRP above the detection threshold limit, only 1 (8.33 %) patient had raised CRP in urine. CRP was raised two weeks after initiation of antifungal therapy in 3 of the above 12 sera and all these 3 cases turned out to be recurrent cases of cryptococcal meningitis. Conclusion Given the high incidence, morbidity and mortality associated with cryptococcal meningitis, it would be ideal if a screening test could be used to exclude this diagnosis based on the presence of biomarkers in serum or urine which would mean less discomfort for the patient in addition to decreased laboratory examination costs.

Women's Mental Health Open Access

Exploring the Mechanism of Complex Lemon-Angelica Sinensis-Boswellia Essential Oil on Anxiety Disorders with Melasma Through Network Pharmacology and Experimental Validation

Dec 2025
Liu LipingCorresponding author

The incidence rate of melasma is notably high among patients with anxiety disorders. Aromatherapy primarily influences the physiological and psychological states of individuals through the inhalation or application of essential oils, thereby facilitating the treatment or alleviation of various conditions. This study aims to explore the action mechanism of complex lemon-angelica sinensis -boswellia essential oil (CEO) in treating anxiety disorders with melasma. We investigated the active ingredients, targets, and pathways of CEO in relation to anxiety and melasma using network pharmacology. We employed cell assays and conducted nebulized essential oil inhalation tests on CUMS mice to validate the intervention effects of CEO on anxiety. A total of 28 active components, including neryl acetate, 3-butenylphthalide and octyl acetate, and 26 cross-targets, such as ESR1, CCND1 and PIK3CA, were identified. GO and KEGG pathway analyses indicated that these cross-targets were primarily involved in endocrine regulation, cell proliferation, and apoptosis, specifically through PI3K/Akt signaling pathway and calcium signaling pathway. The experimental results demonstrated that CEO significantly reduced the secretion of NO, TNF-a and IL-6, as well as the mRNA expressions of ESR1, CCND1 and PIK3CA in cells compared to the inflammatory cell model. Furthermore, CEO notably decreased the forced swimming immobility time of mice and the levels of IL-1β, ESR1 and CCND1 in hippocampus when compared to model mice. These findings suggest that CEO may regulate ESR1, CCND1 and PIK3CA through its citral, 3-butylphthalate and neryl acetate, thereby influencing endocrine function, cell proliferation and apoptosis, inhibiting inflammation and anxiety-like behavior in CUMS-induced mice.

Diagnostic Performance of Smear Microscopy and Xpert MTB/RIF Versus MGIT Culture in Republic of Congo

Nov 2025 DOI 10.14302/issn.2690-4721.ijcm-25-5786
Ornelle Elion Assiana DarrelCorresponding author

In the Republic of the Congo, tuberculosis (TB) remains a major public health concern. Although the GeneXpert MTB/RIF assay is the WHO-recommended first-line diagnostic test, smear microscopy is still used for treatment monitoring and in facilities where molecular testing is limited. Evaluating the diagnostic accuracy of smear microscopy compared to GeneXpert and MGIT culture is essential to guide diagnostic strategies and strengthen TB control in the country. A cross-sectional study was conducted among 92 presumptive pulmonary TB patients at Makelekele Hospital. Sputum samples were analyzed by smear microscopy, GeneXpert MTB/RIF, and MGIT culture. Sensitivity, specificity, positive and negative predictive value were calculated for smear microscopy and GeneXpert, using culture as the reference standard. Culture detected more Mycobacterium tuberculosis than microscopy (49% vs. 32%, P<0.001). Smear microscopy showed a sensitivity of 58% (95% CI: 43–71%) and specificity of 92% (95% CI: 80–97%). GeneXpert detected more MTB (62% vs. 49%, P<0.001) with a sensitivity of 98% (95% CI: 89–100%) and specificity of 72% (95% CI: 58–83%). GeneXpert showed superior sensitivity for TB detection, while microscopy remained specific. Expanding GeneXpert testing across the Republic of the Congo will improve TB management.

Fecal Shedding, Antimicrobial Resistance and In Vitro Biofilm formation on Simulated Gallstones by Salmonella Typhi Isolated from Typhoid Cases and Asymptomatic Carriers in Nairobi, Kenya

Apr 2024 DOI 10.14302/issn.2690-4721.ijcm-24-5030
S. Gunn* JohnCorresponding author

Typhoid fever, caused by the human restricted pathogen Salmonella Typhi, remains a major global public health concern. Even after successful treatment, approximately 3-5% of patients with typhoid fail to clear the bacteria within one year and become chronic carriers. Most typhoid carriers have gallstones in their gallbladder, and biofilm formation on gallstones is highly correlated with chronic carriage. This study’s goal was to identify asymptomatic typhoid carriers in an endemic setting in Kenya, and to compare acute versus chronic isolates. A cohort of typhoid fever patients identified through blood and/or stool culture, and their household contacts, were followed up after treatment to detect longitudinal S. Typhi stool shedding. An abdominal ultrasound scan was used to identify individuals with gallstones. A total of 32 index patients and 32 household contacts were successfully followed-up. Gallstones were detected in 4 cases and 1 household contact. The duration of S. Typhi shedding was significantly longer in individuals with gallstones compared to those without, P<0.001. Eighty-three (83) S. Typhi strains were tested for susceptibility to commonly used antimicrobials and examined by in vitro biofilm formation assays. Out of 37 infected individuals, 32.4% had infections caused by multidrug resistant (MDR) S. Typhi strains and only 18.9% were infected by susceptible strains. Non-MDR strains formed significantly better biofilms in vitro than the MDR strains (P<0.001). This study provides data on S. Typhi chronic carriage that will influence public health approaches aimed at reducing typhoid transmission and the burden of infection.

The Application of Immunoglobulins Immune Response in the Discovery and Development of Safe Therapeutic Agents: A Review Article

Jan 2024 DOI 10.14302/issn.2328-0182.japst-23-4771
Tariku Belay YilkalCorresponding author

Background Immunoglobulins are bio-receptors found embedded in the cell membrane with a biological role that detects the harmful molecules of a test compound. These bio-receptors interface between a biological system and its external environment that transduce information to the effector via intermediate messengers in which its response efficiency usually exhausts at high doses of exposure to external stimuli. The purpose of this review article is, therefore, to elaborate on the computational method for systemic biology which was designed to convert qualitative pharmacological data into the quantitative one that might help to determine the toxicity of a test compound. Methods First, acute toxicity studies using different levels of doses prepared from each test compound have been conducted on Balb c mice. Then, blood specimens from the tail and facial veins of each sampled Balb c mouse were collected 3 days before dosing as a reference test and 4 hr after dosing for comparison. The changes in the efficiency of immunoglobulins immune response (ΔIg) after dosing were determined using quantitative immunoassay and the body’s response against the dose as the toxic reaction rate (r) and the toxic severity (s) were finally determined using computational methods as r=d/t-ΔIg mg/sec and (s=r/w×100) %/sec respectively, where (w) represents the body weight of a study animal, (t) represents the period of time at which undesirable bio-physiological responses manifested on treated study animals and (ΔIg) represents the changes in the concentration of immunoglobulins in blood serum after dosing. Results The results of different studies revealed that the dose has never limited the toxic property of a test compound but the length of time at which the undesirable side effect was manifested on study animals. The period of time at which adverse effects manifested on treated Balb c mice was inversely related to the amount of dose administered in the oral route. The higher the dose of the administered test compound, the shorter the period of time at which the undesirable side effect was manifested on treated Balb c mice. This means that the adverse effect of test compounds was not because of the dose but rather due to its toxic reaction rate which ultimately determined the toxic severity in the natural process of treated Balb c mice. Balb c mice treated with a dose whose toxic reaction rate was ≤ 0 survived from death whereas Balb c mice treated with a dose that had a toxic reaction rate of > 0 died at different lengths of time after dosing depending on the toxic severity of a test compound. It could be a scientific fact to declare that a test compound is safe when the toxic reaction rate (r) and toxic severity (s) of a dose is ≤ 0 and toxic when it is > 0 in the natural processes of a study animal.

Enzymes Open Access

Enzyme Immobilization on Polypropylene Film: A Role Model for Biocatalytic Polymer Membranes?

Dec 2023 DOI 10.14302/issn.2690-4829.jen-23-4799
Gartner PatriziaCorresponding author

Polymer electrolyte membrane (PEM) technologies hold promise for sustainable energy solutions, yet pinhole-related challenges persist. Our research introduces a novel biohybrid approach to self-healing, enhancing multiple healing cycles with minimal membrane disruption. Initial steps involve immobilizing enzymes on a polymeric membrane. This study establishes the immobilization process and analytical framework through enzyme immobilization on polypropylene. Applicability and stability are investigated, laying groundwork for potential Nafion™ applications and advancing climate neutral energy. Qualitative analysis employs colorimetric p-NPA assay on polypropylene-immobilized lipase from Candida rugosa (CRL) and Lipase B from Candida antarctica (CALB). Both enzymes hold their temperature optimum at 50°C which is increased by 10°C via immobilization. Diisopropylcarbodiimide (DIC) is optimal for immobilization. Synchronous enzyme and DIC addition is advantageous. After 8 reuse cycles, immobilized enzymes retain 54.3% residual activity. Immobilizates exposed to PEM fuel cell conditions show better stability due to covalent immobilization than free CRL. Yet, declines occur under stressors like 60 °C and concentrated alcohol. Immobilizates remain resilient at pH 3 and under oxidizing as well as reducing conditions constituted by varied gas atmospheres. Considering PEM fuel cells' operational range, in-depth investigations across conditions are vital. Future studies target long-term PEM fuel cell lifespans, focusing on extremophilic enzymes or modifications for high-temperature stability. Subsequently, the transferability of the immobilization method to Nafion™ shall be deliberated based on the outcomes.

A Review on Monkey Pox: Role of One Health Approach Against Monkey Pox

Oct 2023 DOI 10.14302/issn.2641-4538.jphi-23-4622
Geinoro Alleyo TarikuCorresponding author

The Ortho-poxvirus virus, which causes monkey pox, is a member of the Poxviridae genus. It was initially found in primates. In 1970, the Democratic Republic of the Congo reported the first instance of monkey pox. From there, it spread to a number of countries both inside and outside of Africa. There are two genetic varieties of monkey pox, which have been spread to people through respiratory droplets, and touch with objects contaminated by an affected person and consequently is often encountered in work situations. Among the countries with the worst effects are Nigeria and the Democratic Republic of the Congo. Non-human primates, rats, squirrel, and or mice are just a few of the creatures that can become infected by monkey pox. Although the origins of monkey pox infections are unknown, rodents from Africa and non-human primates like monkeys may contain the viruses and infect humans. Monkey pox is more likely to naturally infect rodents. Monkey pox has symptoms and lesions that are difficult to distinguish from smallpox in its clinical manifestations. Fever, chills, migraines, tiredness, tonia, swollen lymph nodes, back pain, and myalgia are some of the clinical symptoms of monkey-pox. A few examples of diagnostic tests include immune-fluorescent antibody assays, enzyme-linked immune-sorbent assays, and real-time polymerase chain reactions. A specialized vaccine that offers complete protection against by them on key-pox virus exists, yet there is no specific therapy for human monkey infection and interaction with the vaccinia virus. Smallpox vaccination can give cross-immunity with partial protection against infection and a reduction in symptom severity. Unfortunately, community health effects in the view of one health approach has not been addressed in vast. Therefore, the objectives of this review paper are to discuss the community health effects of monkey pox and to emphasize the role of one health approach against monkey pox.

Veterinary Healthcare Open Access

Camel Brucellosis in Ethiopia: Seroprevalence and Associated Risk Factor

Oct 2023 DOI 10.14302/issn.2575-1212.jvhc-23-4532
Abda Neja SultanCorresponding author

Camels are a significant source of income for nomadic populations in many developing countries, including Ethiopia. Camels are well adapted to dry and semi-dry regions, providing income, food security, and transportation. However, camel production and productivity are constrained by infectious diseases, such as brucellosis, which is a highly infectious bacterial disease that affects camels and humans worldwide. Brucellosis causes significant economic losses due to abortion, low herd fertility, and decreased milk production. In Ethiopia, the prevalence of camel brucellosis varies depending on factors related to the host, agent, climate, and management system, with a reported prevalence ranging from 0.5% to 11.9%. Accurate diagnosis of camel Brucellosis is essential for herd-based screening of animals. Although culturing the pathogen is the preferred method for diagnosis, serological tests such as Rose-Bengal plate test (RBPT), Enzyme-linked immunosorbent assay (ELISA), and Complement fixation test (CFT) and polymerase chain reaction (PCR) assays have been developed. Implementing effective diagnosis and surveillance systems to control the spread of brucellosis in animals and humans is very important, on top of awareness campaigns, vaccination programs, and suitable laboratory establishment recommended. Continued research is essential to maintain the health and productivity of camel populations, particularly in pastoral areas where camels play a significant role in the livelihood of communities. Therefore, the present paper views the seropositive prevalence and potential risk factors associated with camel brucellosis in Ethiopia.

Veterinary Healthcare Open Access

A Review of Attempts to Identification and Antifungal Susceptibility of Dermatophytes (Microsporum Canis and Tricophyton Mentagrophytes) Isolated from Infected Cats and Dogs with Experimental Dermatophytosis of Guinea Pigs

Jul 2023 DOI 10.14302/issn.2575-1212.jvhc-23-4510
Youssef SohirCorresponding author

Dermatophytosis affect companion animal’s skin and keratin appendages as cats and dogs, resulting in red, scaly, itchy, bald, and raised patches like ring. The three main groups are Microsporum, Trichophyton and Epidermophyton. This study collected samples of skin scrapping and hairs from 130 cats and 70 dogs, using common mycological approach samples were examined. Antifungal agar disc diffusion and broth microdilution assays were utilized on some of the isolates. Three groups of Guinea pigs (6 in each) were then infected with one isolate of M. canis or T. mentagrophytes fungi, another skin scrapping samples of virulent fungi was isolated on the 7th and 14th days, blood samples were collected at 14th day. Reverse transcription-PCR to detect 98 bp protease gene. Resulting in 45% of cats and dogs tested positive for Microsporum and Trichophyton species. Agar disc diffusion revealed that the antifungal medication griseofulvin was the most effective against tested isolates. The best results for MIC test were griseofulvin (0.98 µg/ml) followed by acetic acid (0.28 µg/ml). Differential leukocytic count of Guinea pigs showed that monocyte levels remained unchanged, while neutrophil and lymphocyte levels had increased. The active (isolates from Guinea pigs skin scrapping) and dormant cells (isolates from keratin free media) were distinguished by Reverse Transcriptase-PCR. Collectively, qPCR is a successive and feasible method for the diagnosis for Microsporum and Trichophyton species.

Detection of Smuggled Genetically Modified Crops and Assessment of its Environmental Impact in the Ethio -Sudan Trans Boundary Area, North West Ethiopia

Apr 2023 DOI 10.14302/issn.2576-6694.jbbs-22-4390
Berhane NegaCorresponding author

Background The genetic material of the genetically modified crop has been altered to develop the necessary insect resistance features by introducing genes from the Bt (Bacillus thuringiensis) bacterium. The objective of this study was to find smuggled GM Bt crops in the Metema farming area and examine its environmental effects. Method An experimental; Completely Randomized Design (CRD) was used to collect crop samples in the study area. The CTAB (Cetyltrimethyl ammonium bromide) technique was used to isolate DNA from all transported samples, and the purity was determined using a Nano Drop spectrophotometer. Conventional PCR with particular primers for different Bt gene events was used to detect the presence of genes. Furthermore, utilizing Bt cotton specific primer sets, the prevalence of GM cotton was measured, and amplified fragments were confirmed using agarose gel electrophoresis. Result The PCR results revealed that 15 (33.3 percent) of the samples were Bt cotton smuggled from Sudan. The PCR assay also revealed the presence of GM maize. Moreover, the effects of GM genes on the environment were studied in diseased samples, and no transgenes were found. Furthermore, domestic and indigenous crops were used to determine horizontal gene transfers of GM genes to other crops, and the transgene was not found in any of the samples analyzed. Conclusion: In the current study, 28 (13.4%) of the 209 (100%) total analyzed samples were GM crops which indicated the presence of unauthorized GM seeds in the study area. Environmental impact studies and horizontal gene transfer data similarly revealed that the Bt gene was not transferred to other crops and had no harmful environmental effects. For a better understanding of the Impact of imported unauthorized GM seeds, more additional detection of GM events should be done by expanding the sampling site and sample types.

Veterinary Healthcare Open Access

Cytokine Expression in Peripheral Blood Mononuclear Cell Cultures Obtained from Cattle with Different Stages of Natural Mycobacterium bovis Infection

Dec 2021 DOI 10.14302/issn.2575-1212.jvhc-21-4034
Fernando Díaz-Otero,Corresponding author CENID-Salud Animal e Inocuidad. Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias (INIFAP). Carretera México-Toluca, Km. 15.5, C.P. 05110, Ciudad de México, México.

In bovine tuberculosis (bTB), cellular, humoral, or both types of immune responses have been observed. The purpose of this study was to examine the immune status of tuberculous cows based on the differential cytokine gene expression associated with Th1 (IFN-γ, IL-2), or Th2 (IL-4, IL-10) responses. Twenty-three (23) cows belonging to a dairy herd located in a rural region of the State of Hidalgo, México, were selected for the study. Single Intradermal Comparative Cervical Tuberculin (SICCT) Test, Interferon-Gamma (IFN-γ) Release Assay (BOVIGAM), and Enzyme-Linked Immunosorbent Assay (ELISA) were used for detection of cattle infected by M. bovis. Thirteen cows were positive to all the tests (Group 1); ten cows were positive only to ELISA (Group 2), and the remaining Group (Group 3, control) included cows negative to all the tests. Peripheral blood mononuclear cells (PBMC) from animals were in vitro stimulated by bovin purified protein derivative (PPD), avian PPD, and Concanavalin A (Con A) mitogen for 72h. Changes in the levels of expression of mRNA of the respective cytokines was measured by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) using β-actin gene as internal control. In group 1, PPD bovis and Con A-stimulated cells exhibited high production of IFN-γ, IL-2 and IL-4, but not IL-10. In contrast, PPD avium-stimulated cells displayed a low production of cytokine transcripts. In group 2, cells showed a significant production of IL-10 in response to bovine PPD (P< 0.001). In the control group, a high production of IFN-γ and IL-2 was observed only in Con A-stimulated cells. Post-mortem examinations in animals of group 1 showed slight and medium lesions in lymph nodes, whereas in group 2, the lesions were more extensive. Results indicate differences on gene expression levels of cytokines considered to determine balance in Th1/Th2 response among the evaluated groups. In addition, high levels of antibodies against M. bovis and high IL-10 expression in PBMC together are indicators of progressive bTB when both tuberculin test and IFN-γ assay are negative in tuberculous anergic cattle. Inclusion of serology and IL-10 cytokine expression in in the diagnosis checklist improves detection of infected cattle to help control bovine tuberculosis.

Serum Ferritin Level as a Prognostic Marker of 30 days In Hospital Mortality of Coronavirus Disease 2019 (COVID 19) Pneumonia at World Citi Medical Center: A Retrospective, Observational Cohort, Single Center Study

Dec 2021 DOI 10.14302/issn.2692-1537.ijcv-21-4025
M. Quirit AllenCorresponding author MD.

Introduction The COVID-19 pandemic continues to affect a large swath of the global population. The Philippine records four hundred seventy-four thousand sixty-four (474, 064) confirmed COVID 19 cases since December 31 2020. The COVID 19 pandemic recently highlighted the role of systemic hyperferritenemia as a major cause of death. In this study, we were able to correlate the serum ferritin level and predict 30 day in hospital mortality in COVID 19 pneumonia. Objective The aim of the study is to investigate the correlation between serum ferritin level and disease mortality in COVID19 pneumonia with subset analysis on demographics and co-morbidities of patients with COVID 19 pneumonia. Methodology We reviewed the records of all laboratory confirmed COVID 19 patients from World Citi Medical Center from April 2020 up to April 2021.A statistically significant sample size of seventy nine (79) admitted patients were used in this study. A serum ferritin level was assayed using electrochemilumenescence immunoassay with a Roche COBAS analyzer. Results Result showed that high ferritin level is associated with in hospital mortality. With ferritin level of 1437.07ng/ml, poor clinical outcome and in hospital mortality was considered. We also observed that demographics and co morbidities of patients in this study were significant to predict in hospital mortality. Further sub-analysis of co morbidities such as Hypertensive cardiovascular disease, Type 2 Diabetes Mellitus, Chronic kidney disease, Liver disease, Chronic obstructive pulmonary disease and Cerebrovascular disease showed poor outcome which were directly related to ferritin levels with p value of <0.0001. Conclusion This study has demonstrated that elevated ferritin levels were shown to correlate with 30 day in hospital mortality as well as medical comorbidities such as Hypertensive Cardiovascular disease, Type 2 Diabetes Mellitus, and chronic kidney disease have shown significant evidence for in hospital mortality.

Ameliorative Potential of Chlorogenic Acid on Rotenone-Induced Neurotoxicity in Drosophila Melanogaster Model

Nov 2021 DOI 10.14302/issn.2641-4538.jphi-21-3993
Adenike Adeyemo-Salami OluwatoyinCorresponding author Nutritional and Industrial Biochemistry Unit, Department of Biochemistry, College of Medicine, University of Ibadan, Ibadan, Oyo State, Nigeria.

Chlorogenic acid (CA), abundantly found in green coffee beans, is a phenolic compound with antioxidant and anti-inflammatory properties amongst others. Exposure to rotenone, a natural pesticide, induces Parkinsonism (a type of neurodegeneration) through the induction of mitochondria dysfunction and oxidative stress. Phytochemicals with antioxidant properties may be promising in attenuating this condition. In this research, the ameliorative role of CA on rotenone-induced toxicity in Drosophila melanogaster was evaluated. Drosophila melanogaster (Harwich strain, 1- 3 days old) was used. 6 groups of five vials each with 50 flies/vial were exposed to CA (0; control (2% ethanol), 7.5, 15, 30, 45 and 60 mg/kg diet) for 28 days in the longevity analysis. A 28-day survival assay was carried out with rotenone (0, 250 and 500 μM). CA (30 mg/kg diet) was selected to evaluate its ameliorative potential on rotenone. For the study, the flies were divided into four groups of five vials each and exposed to CA and rotenone; Group A- control (2% ethanol), Group B- CA only, Group C- rotenone only and Group D- CA (30 mg/kg diet)+ rotenone (500 μM)for 7 days. Thereafter, the homogenate was evaluated for oxidative stress status, rate of emergence, negative geotaxis and acetyl cholinesterase activity. CA (30 mg/kg diet) extended the lifespan of flies by 21.4%. Also, CA ameliorated rotenone-induced perturbation in catalase, glutathione-S-transferase and acetyl cholinesterase activities, total thiol and glutathione levels, and behavioral deficit (p < 0.05). CA may have ameliorative effect against rotenone-induced toxicity and Parkinsonism.

SARS-Cov-2 Viral Kinetics in Mild COVID-19 Patients Treated with Chloroquine Regimens or Standard of Care

Aug 2021 DOI 10.14302/issn.2692-1537.ijcv-21-3924
Siripassorn KrittaechoCorresponding author Bamrasnaradura Infectious Diseases Institute, Nonthaburi, Thailand

This study measures the impact of chloroquine (CQ) therapy in reducing SARS-CoV-2 viral load in infected individuals and hence its transmissibility by describing changes in nasopharyngeal SARS-CoV-2 RNA kinetics in patients receiving standard of care (SOC) or CQ +/- ritonavir-boosted lopinavir (LPV/r). The nasopharyngeal (NP) samples were collected from mild COVID-19 patients admitted at Bamrasnaradura Infectious Diseases Institute between March and April of 2020. These patients either received SOC, or a high dose of CQ with loading dose, or high dose of CQ plus LPV/r. The samples were tested at AFRIMS using a quantitative RT-PCR assay. Levels of CQ in the plasma were measured 6 days post initiation of their treatment. In some instances, viral isolation was attempted to determine SARS-CoV-2 viability. Analyses of the clinical outcomes showed that CQ +/- lopinavir did not contribute significantly to decreasing the number of days with detected SARS-CoV-2 RNA. Viral NP GEs declined faster in the CQ group, but benefits diminished rapidly with delays in treatment initiation. Funding Global Emerging Infections Surveillance, Armed Forces Health Surveillance Branch (GEIS-AFHSB) for all research-related activities at the AFRIMS

Nephrology Advances Open Access

Evaluation of Renal and Cardioprotective Potential of the Biofield Energy Treated Proprietary Test Formulation on L-NAME and High Fat Diet-Induced Cardiovascular Disorders in Sprague Dawley Rats

Jun 2021 DOI 10.14302/issn.2574-4488.jna-21-3847
Jana SnehasisCorresponding author Trivedi Science Research Laboratory Pvt. Ltd., Thane (W), Maharashtra, India.

The aim of this study was to evaluate the impact of Biofield Energy Treated/Blessed Proprietary Test Formulation and Biofield Energy Treatment/Blessing per se on kidney biomarkers on L-NAME and high fat diet (HFD)-induced cardiovascular disorders in Sprague Dawley rats. In this experiment, the functional kidney biomarkers such as epinephrine/adrenaline, inducible nitric oxide synthase (iNOS), angiotensin-II, C-reactive protein (CRP), and renin were measured using ELISA assay. A test formulation was formulated including minerals (magnesium, zinc, copper, calcium, selenium, and iron), vitamins (vitamin C, vitamin B6, vitamin B12, vitamin B9, and vitamin D3), cannabidiol (CBD) isolate, Panax ginseng extract, and β-carotene. The components of the test item were divided into two; one section was defined as the untreated test formulation, while the other part and three group of animals received Mr. Mahendra Kumar Trivedi’s Biofield Energy healing/Blessing remotely for about 3 minutes. The results showed that the level of adrenaline was reduced by 31.62%, 19.58%, 34.32%, 37.07%, and 29.87% in the G5 (L-NAME + HFD + the Biofield Energy Treated test formulation), G6 (L-NAME + HFD + Biofield Energy Treatment per se to animals from day -15), G7 (L-NAME + HFD + the Biofield Energy Treated test formulation from day-15), and G8 (L-NAME + HFD + Biofield Energy Treatment per se plus the Biofield Energy Treated test formulation from day-15), and G9 (L-NAME + HFD + Biofield Energy Treatment per se animals plus the untreated test formulation) groups, respectively as compared to the disease control group (G2). Moreover, the level of iNOS was reduced by 56.76%, 49.51%, 61.79%, 57.63%, and 62.44% in the G5, G6, G7, G8, and G9 groups, respectively, as compared to the disease control group (G2). Additionally, the level of angiotensin-II was decreased by 41.09%, 34.92%, 60.65%, 53.28%, and 60.09% in the G5, G6, G7, G8, and G9 groups, respectively, as compared to the G2 group. The level of CRP was decreased by 47.21%, 38.89%, 59.81%, 55.52%, and 64.02% in the G5, G6, G7, G8, and G9 groups, respectively as compared to the G2 group. Besides, the level of renin was decreased by 20.27%, 20.13%, 12.99%, and 25.73% in the G5, G7, G8, and G9 groups, respectively as compared to the G2 group. Overall, the data suggested significance improvement of vital functional kidney biomarkers of the Biofield Energy Treated/Blessed test formulation and Biofield Energy Treatment per se along with preventive measure on the animal with respect to various pathological conditions that might be beneficial various types of cardiovascular disorders. Therefore, the results showed the significant slowdown the inflammation-related cardiovascular disease progression and its complications/symptoms in the preventive Biofield Energy Treatment group per se and/or Biofield Energy Treated/Blessed Test formulation groups (viz. G6, G7, G8, and G9).

Effect of Wheat Germ, Wheat Germ Oil on Lipid Profile of Hypercholesterolemic Rats

Dec 2020 DOI 10.14302/issn.2835-513X.ijl-20-3247
Z A Soliman GhadaCorresponding author Prof Biochem, NNI, Cairo, ARE

Hypercholesterolemia plays an important role in atherosclerosis and cardiovascular diseases (CVD) that represent one of the greatest worldwide medical problems nowadays. Recently, an increased attention/ interest for natural antioxidant/ hypercholesterolemia as WG/WGO is increasing. Wheat germ/ Wheat germ oil (WG/WGO) is an excellent source of essential and polyunsaturated fatty acids and vitamin E. It is one of the richest natural sources of tocopherol. This work aims to evaluate the effect of administration of wheat germ (WG) /wheat germ oil (WGO) for 6 weeks on serum lipid profile, lipid peroxidation (liver malondialdehyde) in hypercholesterolemic rats. One hundred and twenty maleSprague Dawelyrats weighing 180-192 gm were used in this study. They were randomly distributed into six groups (20 rats/ group) as follow: g1: normal control, G2: hypercholesterolemic rats; G3: Normal rats treated with WG; G4: hypercholesterolemic rats treated with WG; G5: Normal rats treated with WGO; G6: hypercholesterolemic rats treated with WGO for 6 weeks. Also WG was analysed for its nutritive value, while WGO was analysed for fatty acid profile, and studied for its physico-chemical properties. The results showed significant elevation of liver malondialdehyde (MDA) and serum lipid profile in untreated rats fed on hypercholesterolemic diet. Hypercholesterolemic rats treated with WG/WGO showed an improvement in the biochemical assay of their lipid profile compared with untreated hypercholesterolemic rats. WG/WGO may be able to protect against atherosclerosis and CVD. The administration of WG /WGO to diets of rats caused a marked reduction in TC, LDL-C, VLDL-C, TG and MDA. This improvement effect may be mediated via enhancement of the antioxidant defence system and other factors. However, further clinical studies on human beings are required to assess the efficacy and safety of the WG/WGO.

Clinical Evaluation of Significance of 25(Oh)D (Vitamin D) Status in Swine Flu (H1N1)

Sep 2020 DOI 10.14302/issn.2379-7835.ijn-20-3369
Kishan Gupta BalCorresponding author Senior Professor, In-charge Medical ICU, Department. of Medicine, S.P.Medical College, Bikaner.

Background Since swine flu has been declared pandemic in 2009 it has become a major challenging public-health problem associated with high morbidity and mortality. 25(OH)D deficiency is also pandemic and has been reported to be clinically correlated with decreased immunity and respiratory infections. The possible role of vitamin D in infections is implied from its impact on the innate and adaptive immune responses. This study is planned to evaluate clinical significance of 25(OH)D status on course and outcome in hospitalized cases of swine flu and to compare it with normal healthy subjects living in the same vicinity to evaluate if vitamin D is having any protective effect. Material & Methods Present prospective cross-sectional study was conducted on 79 RT-PCR confirmed cases of swine flu admitted during recent epidemic. All patients were evaluated thoroughly by clinical history physical examination and laboratory investigations as per Performa and followed-up during hospital stay. 25-hydroxyvitamin D (25(OH)D) estimation was done by electro-chemiluminescent Assay in all the cases and it was also done in 36 normal healthy family members of study patients living in the same vicinity (control group). Results High prevalence (70.9%) of low (≤30ng/ml) status of 25(OH)D was observed in cases of swine flu as compared to control group. 25(OH)D status was associated with severity of illness. Mean value of 25(OH)D in mechanically ventilated patients was 9.81±6.43 while it was 22.76±11.35 ng/ml in patients who do not required ventilation (p<0.05). Mean 25(OH)D level in patients who stayed in hospital for <5 days was 28.60±8.79 ng/ml, 24.18±11.67 for 6-10 days and 8.23±2.12 for >10 days (p<0.01). Mean value of 25(OH)D in patients who died was 9.59±5.90 ng/ml as compared to 23.13±11.62 ng/ml who survived (p<0.01). Conclusion Our study suggests that 25(OH)D may have preventive role for swine flu infection. Low level of 25(OH)D is associated with high morbidity in terms of increase requirement for mechanical ventilation, multiorgan dysfunction and long duration of hospital stay. 25(OH)D deficiency is associated with high mortality in swine flu. 25(OH)D status should be given due consideration in high risk patients especially during winter season.

Effects of Selected Secondary Metabolites in Leaf Extract of Jatropha Tanjorensis on Some Gonadal Hormones in Male Wistar Rats

Jul 2020 DOI 10.14302/issn.2576-6694.jbbs-20-3466
John AkighirCorresponding author Department of Biochemistry, College of Science, Federal University of Agriculture, Makurdi P.M.B. 2373 (970001) Nigeria

Background and Objective The use of medicinal plants in industrialized societies for extraction and development of many drugs and other chemotherapeutics and traditionally for herbal remedies has increased in recent times. Plant–based medicine is essential in health care services with about 80% global population relying on it because of its cheap source and availability. Jatropha tanjorensis is one such plant used by males and females of childbearing age for treatment of reproductive problems such as infertility. Literature on isolation and characterization of the secondary metabolites in this plant may not be common. Against this backdrop, this research work was carried out to isolate, characterize and determine the effects of J. tanjorensis on the gonadal hormones of male wistar rats. Materials and Methods The secondary metabolites were isolated, characterized, and identified using nuclear magnetic resonance. The experiment was conducted using 25 male wistar rats weighing between 180-200 g randomized into 5 groups, 3 controls and 2 treatment groups of 5 rats each. The treatment groups received 25 mg/kg body weight of phytol and lupeol orally by gastric lavage for 14 days. The animals were anaesthetized and blood samples collected for hormonal assay. Result The experimental data was analyzed using one-way analysis of variance (ANOVA) with Statistical Package for Social Sciences (SPSS) version 17.0, while the post hoc test assessed using Duncan Multiple Range Test at p ≥ 0.05. There was a significant decrease (p ˂ 0.05) in the levels of FSH, LH and TST in the treatment groups when compared to the control groups. The motility and sperm count decrease significantly (p ˂ 0.05) when treatment groups were compared to the control animals. The secondary metabolites, phytol and lupeol present in the leaf extract of Jatropha tanjorensis were responsible for the decrease in some of the gonadal hormones studied.

Efficacy of Phytochemical Constituents of Castor Essential oil Towards the Mucor-Mycotic Mold Cunninghamella Bertholletiae

Jul 2020 DOI 10.14302/issn.2377-2549.jndc-20-3484
Shakeel Iqubal S.M.Corresponding author Department of General Science, Ibn Sina National College of Medical Sciences, Al Mahajar Street: 31906, Jeddah 21418, Kingdom of Saudi Arabia.

The aim of this experiment is to study the efficacy of phytochemical constituents of Castor essential oil towards the mucor-mycotic mold Cunninghamella bertholletiae.The standard chemical analytical methods were used for the rapid study of the phytochemical constituents responsible for the antimicrobial efficacy of the procured castor essential oil. The standard antimicrobial assay technique employed to study the comparative values of the efficacy of the procured castor essential oil with that of the standard antifungal chemical agents against the clinical isolates obtained from the immune suppressed patients samples of Cunninghamella bertholletia mold mucor-mycotic infections. The best susceptibility values recorded in the standard antifungal agents against the clinical isolates of Cunninghamella bertholletiae was with Amphotericin B showing the average zone of inhibition diameter of 20.66 mm with the average MIC value of, 1.66 (µ/ml) but the antimicrobial assay results for the Castor essential oil showed better values with an average disc diffusion of 22.44mm zone of inhibition diameter with average MIC value of 1.72 µ/ml .This study has shown that the phytochemical compounds present in the Castor essential oil proves to be more an effective alternative antifungal substance towards the clinical isolates of Cunninghamella bertholletiae.

Antioxidant Activity Open Access

Antioxidant Activity of Surinamese Medicinal Plants with Adaptogenic Properties and Correlation with Total Phenolic Contents

Jul 2020 DOI 10.14302/issn.2471-2140.jaa-20-3478
R.A. Mans DennisCorresponding author Department of Pharmacology, Faculty of Medical Sciences, Anton de Kom University of Suriname, Paramaribo, Suriname

Plant-based preparations are commonly used in Suriname (South America) as adaptogens. In this study, fifteen alleged adaptogenic Surinamese plants have been assessed for their antioxidant activity (AA), total phenolic contents (TPC), and total flavonoid contents (TFC). The investigated plants were Anacardium occidentale, Spondiasdulcis, Annona muricata, Euterpe oleracea, Oenocarpus bacaba, Luffa acutangula, Punicagranatum, Malpighia emarginata, Syzygiumaqueum, Syzygiumcumini, Averrhoa carambola, and Renealmiaalpinia (fruit); Hibiscus sabdariffa (calyx); as well as Aloe vera and Cestrum latifolium (leaf). Aqueous extracts (1 - 3,000 μg/ mL) were prepared. AA was determined by the FRAP and the DPPH assay. TPC and TFC were determined by the Folin-Ciocalteu’s and an AlCl3 colorimetric method, respectively, using gallic acid (GA) and rutin (R), respectively, as standards. Data are means ± SDs (n ≥ 3; P < 0.05). FRAP values and DPPH-scavenging activities correlated positively with each other and with TPC but not with TFC. The preparations from M. emarginata, A. carambola, A. occidentale, O. bacaba, C. latifolium, and H. sabdariffa displayed the highest FRAP values (54 ± 14 to 412 ± 30 µM Fe2+/100 μg), DPPH-scavenging activities (IC50 values of 33 ± 14 to 250 ± 50 μg/mL), and TPC (51 ± 4 to 280 ± 78 µM GAE/100 µg). TFC of all samples were ≤ 10 ± 3 RE/100 µg. The adaptogenic properties of these plants may (partially) be attributed to their high content of antioxidant phenolic compounds and may make them candidates of novel sources of health-promoting antioxidants.

DNA And RNA Research Open Access

Molecular Study of Hepcidin HAMP (-582A/G) Gene Polymorphisms and Measurement of Serum Hepcidin Level among Sudanese Patients with Anemia of Chronic Kidney Disease

May 2020 DOI 10.14302/issn.2575-7881.jdrr-20-3343
Hussen Abdelrhman AmgedCorresponding author Assis Professor, Department of Hematology and Immunohematology, Omdurman Islamic university / Sudan

Background Anemia of chronic disease is anemia found in certain chronic disease states, is typically marked by the disturbance of iron homeostasis or hypoferremia. Chronic renal failure is currently known as Chronic Kidney Disease (CKD) or Chronic Renal Insufficiency (CRI) implies long-standing, progressive and irreversible renal parenchyma disease resulting in diminished renal function up to 40 to 60%. Often, chronic kidney disease is diagnosed as a result of screening of people known to be at risk of kidney problems, such as those with high blood pressure or diabetes and those with a blood relative with chronic kidney disease. This disease may also be identified when it leads to one of its recognized complications such as cardiovascular disease, anemia, or pericarditis.                             Methods Sysmex kx21 used to CBC and the Cobase411 used to iron profile. Enzyme-Linked immunoassay (ELISA) was used to determine the level of serum hepcidin.  Sample preparation and PCR detection of HAMP DNA Polymorphisms: Restriction digestion of PCR products was done using Fast Digest. (Figure 1).                                                                                         Results Serum hepcidin levels higher in patients with anemia of chronic kidney disease compared with healthy controls mean. The polymorphisms of the hepcidin gene promoter in Sudanese patients with ACKD showed that the hepcidin HAMP AA genotype 70, AG 23, and GG 7 in 100 patients dialysis-dependent and AA 83, AG 17 and GG 0, and the allele A are more frequent in patients affected by ACKD. Significant statistical association observed between the hepcidin level and end-stage kidney disease. Conclusion This study evaluates for the first time the association between anemia of chronic kidney disease and hepcidin genes promoter polymorphisms and show that the hepcidin HAMP AA genotype and the allele A are more frequent in patients affected by ACKD, further investigation is needed, our data support the hypothesis and hepcidin HAMP are important in the pathophysiology of ACKD.

Effect of Hexavalent Chromium on the WBCs of the Fresh Water Fish, Labeo rohita

May 2020 DOI 10.14302/issn.2641-7669.ject-20-3313
A.J ThatheyusCorresponding author PG & Research Department of Zoology, The American College, Madurai, Tamil Nadu, India.

Industrial effluents containing heavy metals may reach aquatic systems either through direct discharge or surface runoff and cause damage to aquatic organisms affecting their immune system and health. Hence the present study has been undertaken to observe the effects of hexavalent chromium on the WBCs of the fresh water fish, Labeorohita. WBCs play a major role in the immune response of the fish. For acute toxicity determination, healthy fish were subjected to static bioassays. The 24, 48, 72, and 96hr LC50 values were 50.88, 42.03, 28.09 and 10.87 ppm respectively. The fish were exposed to 0.5, 1, 1.5 and 2 ppm for 20 days. Differential count of WBCs and total WBC count were determined after every five days for twenty days. Lymphocytes exhibited a decline while the other cells and total WBC count exhibited an increase due to hexavalent chromium exposure. The results were subjected to two way analysis of variance.

Invivo Impact of Malaria and HIV Co-Infection on CD4 Cell Count of Infected Patients of Niger Delta Extraction

May 2020 DOI 10.14302/issn.2328-0182.japst-20-3347
Obioma AzuonwuCorresponding author Department of Medical Laboratory Science, Medical Bacteriology / Virology / Parasitology Unit, Rivers State University, Nkpolu – Oroworukwo, Port Harcourt, Rivers State, Nigeria.

The study evaluated the impact of co-infection of malaria parasitaemia, and HIV positive indices on the CD4 cell count of 120 HIV infected subjects, who were already diagnosed and visiting Braithwaite Memorial Specialist Hospital Port Harcourt for routine Medical check-up. Also, a control group of 40 HIV negative were included as part of the study control group. The subjects were between the age ranges of ≤10–79 years respectively. A double check laboratory assay was conducted to detect the presence of antibody to HIV as confirmed using immunocomb 11 and Determine for HIV status. A thick Blood film stained with field stain (A and B) was used to detect the presence of malaria parasite in the subject’s blood. Furthermore, CD4 cell count was assayed using Partec cyflow counter (Partec, Germany). Excel and Graphpad statistical software were used for analysis of the data generated. The result among the HIV positive subjects and control subjects revealed that the highest positive for malaria infection was observed among ≤10 years age group as 2 (100%) and 11 (84.61%) respectively. In the HIV positive subjects, the distribution of malaria infection among sex revealed a high rate in male 42(77.78%) than in female 44 (66.67%). Similarly, the control recorded a high rate of malaria infection in male 11 (57.89%) than in female 7 (33.33%). However, 86 (71.67%) had malaria and HIV co-infection while 34 (65%) had only HIV mono infection. The positive HIV subjects who had CD4 cells count below 200 cells/mm3 were 15%, above 200-499cells/mm3 were 58.3% while 500 cells/mm3 and above had normal CD4 cells counts for 26%. Nonetheless, for the control subjects, no CD4 cells count of below 200cells/ mm was observed, 2.5% fell within the moderate category while 75% had normal CD4 cells count. Statistical analysis using ANOVA and t-test showed that there is significant difference between CD4 of seropositive and seronegative subjects infected with or without malaria (p=0.00). In addition, a t-test further demonstrated Comparison of Mean CD4 Cell Count among HIV and Malaria Infected and Non-Infected Subjects. MP/HIV Co-Infection and Mono Infection with No Infection showed strong mean difference (p=0.00) in the various CD4 counts while HIV Mono-Infection and others only had a non significant (p=0.44) mean difference between HIV Mono-Infection and No HIV or Malaria Infection. A robust and effective malaria and HIV control management programme should be strongly underpinned; so as to improve the quality of life of patients and HIV patients should be encouraged to live a healthy life style, through the provision of antiretroviral drugs and regular health education engagement, even as the provision of antimalarial treated net would be helpful to the subjects.

RETRACTED: Assessment of the Risk of Hemochromatosis in Polytransfused Sickle Cell Patients at the Abidjan Transfusion Therapy Unit

Apr 2020 DOI 10.14302/issn.2372-6601.jhor-20-3189
Mamadou Sekongo YassonguiCorresponding author Department of Training and Research, National Blood Transfusion Center; Abidjan; Côte D’Ivoire

This article has been retracted on 29 January 2021. VIEW THE RETRACTION NOTICE (https://doi.org/10.14302/issn.2372-6601.jhor-25-5854) In Côte d'Ivoire, sickle cell disease affects 14% of the population. It is responsible for significant morbidity and mortality. Transfusion is a significant element in the management of major sickle cell anemia, which exposes them to post-transfusion hemochromatosis. The biological diagnosis is based on the determination of serum iron and the transferrin saturation coefficient (CST). As the determination of the CST was not available in our exercise context in Côte d'Ivoire, we determined only the ferritinemia. The interest of this work lies in the therapeutic implication linked to the identification of patients at risk of hemochromatosis because chelators are difficult to access for most patients. This was a prospective, descriptive and analytical study, on polytransfused sickle cell patients, followed at the transfusion therapy unit (UTT) of the CNTS of Abidjan, from 2010 to 2018. We included 78 sickle cell patients, all ages and genders who have received at least ten transfusions. The ferritinemia assay was carried out by ELISA. Transfusion exchange, with 59% of cases, was the most used mode of transfusion. The mean ferritinemia was 1719.19 ng / ml. Hyperferritinemia was found in 63% of patients. Most of the patients were on a long-term transfusion program with an average of 27.5 bags of red blood cell concentrates. Thirty-two patients had received at least 20 bags of red blood cell concentrates. We noted 21 patients treated, including 3 with deferoxamine and 18 treated with oral deferasirox. We have identified 33 sickle cell anemia patients at risk for hemochromatosis. The determinants of the risk of hemochromatosis were the high number of blood bags and the method of transfusion.

Auricularia Polytricha (Mushroom) Regulates Testicular DNA Expression and Oxidative Stress Markers of Streptozotocin-Induced Diabetic Male Wistar Rat

Mar 2020 DOI 10.14302/issn.2379-7835.ijn-20-3175
Cyril Abang AgborCorresponding author Department of Anatomy, Collage of Basic Medical Sciences, University of Calabar, Nigeria

Local Nigerian men have been using AuriculariaPolytricha as a treatment for sexual dysfunction without supporting evidence from scientific experiments. This study was to investigate the effect of ethanolic extract of A. Polytricha on testicular DNA expression and some oxidative stress markers using STZ-Induced diabetic rats as a model. The experiment included six groups, Group A (Normal Control, treated with normal saline), Group B (treated with 65mg/kg.bw of STZ), Groups C, D, and E (treated with 250mg/kg.bw, 500mg/kg.bw, 1000mg/kg.bw AP after inducing diabetics), and Group F (treated with 40mg/kg.bw metformin after inducing diabetics). The experiment lasted for 35 days. After termination of the experiment, Fuelgen nuclear reaction was used for DNA demonstration to assess testicular DNA distribution while serum Superoxide Dimutase (SOD), Catalase and Melondialdehyde where evaluated using reagent based antioxidant enzyme assay. Results reveals that SOD and Melondialdehyde activities were remarkably (p<0.05) higher in diabetic control animals when compared with the normal control group. Values in Groups C, D and F that were administered with 250, 500mg/kg.bw A. polytricha and metformin respectively were also significantly (p<0.05) increased when compared with the normal control group. However, diabetic animals placed on 1000mg/kg.bw A. polytrichadid not show any statistical significance in comparison with normal control group but was remarkably (p<0.01) decreased when compared to the diabetic group that received low dose A. polytricha, an indication that the reversal is dose dependent. Catalase concentration in diabetic control animals was remarkably (p<0.05) higher when compared to the normal control but was not significantly (p<0.05) different in groups D (DM+500mg/kg.bw A. polytricha) and E (DM+1000mg/kg.bw A. polytricha) when compared with the normal control group. Diabetic control animals showed reduced magenta colour intensity of DNA and increased clustering and cross linking of DNA strands when compared with the normal control. However the degree of cross link in DNA strands was reduced in the diabetic animals placed on 1000mg/kg.bw A. polytrichawhen compared with the diabetic control group. Reversal in DNA damage and values of serum oxidative stress markers following administration of graded doses of A. polytricha could be attributed to essential phytochemical and therapeutic constituents in A. polytricha like polyphenol and flavonoid which can be found useful in prevention and treatment of diabetes induced testicular dysfunction. In summary, AP can contribute to a reversal in DNA damage and levels of serum oxidative stress markers in treating diabetes-induced testicular dysfunction.

The Effect of 2,4 Dimethylamine salt on the Blood, Liver and Muscle of Oryclotagus Cuniculus

Mar 2020 DOI 10.14302/issn.2637-6075.jpae-20-3198
C. Izah SylvesterCorresponding author Department of Biology, Bayelsa Medical University, Yenagoa, Bayelsa State, Nigeria.

Aminoforce containing 720g/l of 2,4-dimethylamine salt induced changes on some enzymes and electrolytes in the male Oryclotagus cuniculus (New Zealand rabbit) were assayed. The organisms were exposed to varying sub-lethal concentrations of the toxicant (720g/l). The concentrations were prepared by pipetting 0.4mls, 0.8mls and 0.12mls making it up to 1.5L clean water in a metal container to make 2.0 mgl-1, 4.0 mgl-1 and 6.0 mgl-1. Aspartate amino transferase (AST), Alanine amino transferase (ALT) and Acid phosphatase (ACP) were assayed in the liver and blood. Results showed that aspartate amino transferase values in the liver and blood were significant (p<0.05) across the concentration of the toxicants. Aspartate amino transferase increased as the concentration of the toxicant increased in the liver, and decreased as the toxicant concentration increased in the blood. Alanine amino transferase in the blood and liver were akin to AST while ACP values increased in the blood and decreased in the liver as the concentration of the toxicant increased. Electrolytes (Sodium (Na+), potassium (K+) and magnesium (Mg2+) ions) showed statistical deviation across the various concentration of the toxicants. Chloride ion values stabilized in the experimental group being not significantly different (p>0.05) across the various concentration of the toxicants. From the study, AST, ALT and ACP are suitable biomarkers for showing sub-lethal effect of aminoforce on Oryclotagus cuniculus. The effects recorded clearly unveiled the potential effect of this xenobiotics on Oryclotagus cuniculus. Therefore, exposure of Oryclotagus cuniculus to this toxicant will affect the organism’s physiological responses and over prolong period of time it could lead to death. Additionally, via food chain man may be affected. The use of this toxicant close to rabbittory should be done with utmost caution.

Interest of Confirmation Tests in the Diagnosis of Viral Hepatitis C to Blood Donors in Abidjan-Côte d'Ivoire

Jan 2020 DOI 10.14302/issn.2372-6601.jhor-20-3186
Mamadou Sekongo YassonguiCorresponding author Department of Training and Research, National Blood Transfusion Center; Abidjan; Côte D’Ivoire

Introduction The anti-HCV RIBA test verifies the presence of anti-HCV serum antibodies detected by the Elisa test. In Côte d'Ivoire, screening for hepatitis C is done exclusively by enzyme immunoassays. In order to reduce the number of HCV positive blood donor exclusions on ELISA, we conducted this study which aimed to demonstrate the value of the RIBA test in confirming diagnosis of viral hepatitis C to blood donors. Methods Our study, which took place from 02 to 23 February 2008 in the laboratory of Abidjan NBTC, focused on 200 sera of blood donors anti-HCV positive (Elisa test) selected according to the ratio. The DECISCAN HCV PLUS confirmation test of BIORAD was used. Results Among the 200 HCV samples positive by EIA, 49% (98/200) were confirmed positive. RIBA gave an indeterminate result in 40% of cases (80/200); and negative in 11% of cases (22/200) corresponding to false ELISA devices. In RIBA 96 samples had a low ELISA ratio of which 21% (20/96) were RIBA negative, and 79% (76/96) were indeterminate. RIBA positive samples (98/200) had a high ratio in 82% of cases (80/98). The presence of NS3 (C33) and NS4 (C100) was noted in 100% of cases (98/98, C2 in 37% (36/98) of cases and C1 in 18% of cases (18/98). RIBA indeterminate noted the presence of NS3 in 98% of cases (78/80) and NS4 in 30% of cases (24/80). Proteins C1, C2 and NS4 are essential for the diagnosis of confirmation of viral hepatitis C by RIBA. Conclusion These results attest to the lower specificity of enzyme immunoassays (ELISAs); hence the benefit of using RIBA confirmatory tests. A significant number of donors are excluded from blood donation in Côte d'Ivoire on the basis of false positive results obtained by the ELISA technique.

Effect of the Biofield Energy Treated Proprietary Test Formulation for Sleep Biomarkers in the Unpredictable Chronic Stress (UCS) Animal Model 

Jan 2020 DOI 10.14302/issn.2474-9273.jbtm-19-3157
Jana SnehasisCorresponding author Trivedi Science Research Laboratory Pvt. Ltd., Thane (W), Maharashtra, India.

Sleep biomarkers in brain such as melatonin, BDNF (Brain-derived neurotrophic factor), PGD2 (Prostaglandin D2), leptin, orexin-A, and acetylcholine were evaluated in the unpredictable chronic stress (UCS) rodent model in the presence of Consciousness Energy Healing Treated (the Trivedi Effect®) novel test formulation in male Sprague Dawley (SD) rats using ELISA assay. The test formulation was consisted of minerals (Zn, Fe, Cu, Se, Ca, Mg), vitamins (C, E, B6, B12, D3), β-carotene, ginseng, and cannabidiol isolate. The test formulation constituents were divided into two parts, one part of each ingredient was distinct as the untreated test formulation, while the other portion of the test formulation and a group of animals received Biofield Energy Healing Treatment by a renowned Biofield Energy Healer, Mr. Mahendra Kumar Trivedi. The level of melatonin in groups viz. G5 (Biofield Energy Treated Test formulation) and G7 (15-days pre-treatment of Biofield Energy Treated Test formulation) was significantly increased by 17.6% (p≤0.01) and 16%, respectively as compared with the disease control group (G2). Brain-derived neurotrophic factor (BDNF) level in brain was increased by 5.2% in G7 group as compared with the G4. Prostaglandins D2 (PGD2) level was significantly (p≤0.001) increased by 12.7%, 18.1%, 23.7%, and 30.7% in the G6, G7, G8 (15 days pre-treatment of Biofield Energy Treated Test formulation to the Biofield Energy Treatment per se rats), and G9 (untreated test formulation to the Biofield Energy Treatment per se to the rats) groups, respectively as compared with the G2. The level of leptin after Biofield Energy Treatment and with the test formulation was altered. However, orexin-A level was significantly decreased by 37.1% (p≤0.05), 32.6%, 40.5% (p≤0.05), 44.4% (p≤0.05), and 28.2% in the G5, G6, G7, G8, and G9 groups respectively, as compared with the G2. Similarly, acetylcholine (Ach) level was significantly (p≤0.001) decreased by 42.5%, 49.2%, 40.1%, 47.9%, and 45% in the G5, G6, G7, G8, and G9 groups, respectively as compared with the G2. Overall, the results showed the significant slowdown the stress-related disease progression and its complications/symptoms in the preventive in the Biofield Energy Treatment group per se and/or Biofield Energy Treated Test formulation groups (viz. G6, G7, G8, and G9) comparatively with the disease group. The Trivedi Effect® showed increased level of melatonin and decreased levels of insomnia related brain biomarkers which might be helpful to induce better sleep in human.

In Vitro Assessment of Antioxidant Enzymes, Phenolic Contents and Antioxidant Capacity of the Verdolaga (Portulacaceae)

Jan 2020 DOI 10.14302/issn.2379-7835.ijn-19-3144
Q. Almulaiky YaaserCorresponding author Chemistry Department, Faculty of Sciences and Arts, University of Jeddah, Khulais, P.O. Box 355, Khulais, 21921, Saudi Arabia

In this study, the antioxidants and photosynthetic compounds of Verdolaga were examined. Compounds were extracted from distinctive segments of the verdolaga using various solvents such as methanol (40, 60, 80%), ethanol (40, 60, 80%), acetone (40, 60, 80%), and deionized water. The use of 80% methanol led to the highest extracted concentration of phenolic substances and flavonoids. The extracted products (Leaves, Stem strips, and Root strips) were evaluated for their radical scavenging capabilities with DPPH (IC50= 22.26, 20.56, and 32.10), and ABTS (IC50= 2.86, 3.70, and 5.24), reducing power (EC50= 15.70, 16.39, and 21.69), and peroxide scavenging activity (1C50= 1.717, 2.937, and 3.255), respectively. The extracted products were analyzed by a gas chromatography-mass spectrometer. Peroxidase, catalase, and polyphenol oxidase assays were completed for the crude extract of verdolaga’s leave, stem strips, and root strips. As indicated by these tests, extracts of the verdolaga’s roots, stems and leaves using 80% methanol yielded high antioxidant activity. The most elevated concentrations of extracted chlorophyll, lycopene, and carotenoids were from the leaves and the highest concentration of extracted tannin was noted from strips of stems. The highest measures of peroxidase and polyphenol oxidase were identified in root strips and the highest units of catalase was identified in leaves.

Impact of Aluminum Phosphide on the Transferases in Liver and muscle of Parophiocephalus obscurus

Oct 2019 DOI 10.14302/issn.2637-6075.jpae-19-3022
Chibueze Izah SylvesterCorresponding author Department of Biological Sciences, Bayelsa Medical University, Yenagoa, Bayelsa State, Nigeria.

This study assessed the effect of aluminum phosphide on transferases in liver and muscle of Parophiocephalus obscurus (with mean weight of 42.20±1.5 gSD and mean length of 16.50± cmSD, respectively). The fish were obtained from a private fish farm in Yenagoa Metropolis, Nigeria, and the fish was allowed acclimatized to laboratory condition for 7 days, and then exposed to sublethal concentrations (0.00mg/L, 4.20mg/L, 6.30mg/L and 8.40mg/L) of aluminum phosphide for 14 days. Renewal bioassay was adopted in this study. At the end of the experimental period, the fish was dissected and the muscle and liver were collected, processed and analyzed for alanine aminotransferase and aspartate aminotransferase using colorimetric method. Results of the phosphatase at 0.00mg/L, 4.20mg/L, 6.30mg/L and 8.40mg/L were 94.50±6.44µ/L, 134.47±15.27 µ/L, 106.47±9.21 µ/L and 31.00±3.46 µ/L, respectively (liver), 107.50±9.24, 92.00±6.93 µ/L, 116.50±8.95 µ/L and 146.33±9.33 µ/L respectively (muscle) for aspartate aminotransferase; and 40.00±1.15µ/L, 26.50±3.18µ/L, 14.50±2.02µ/L and 9.80±1.44 µ/L, respectively (liver) and 17.00±1.75µ/L, 8.50±0.87µ/L, 21.00±2.89µ/L and 5.50±0.87 µ/L, respectively (muscle) for alanine aminotransferase. Statistically, there were significant variations (p<0.05) among the various concentration in the transferances. In addition, at some concentration, there was significant variations (p<0.05) between the level of the transferases in the muscle and liver. The significant alteration observed in the various concentrations is an indication that aluminum phosphide is lethal to fish. Therefore, caution should be exercise during the use of aluminum phosphide near biological system.

Changes in Adult Rats’ Testis structure Induced by Hypothyroidism and Alleviating Role of L-Carnitine

Sep 2019 DOI 10.14302/issn.2577-2279.ijha-19-3026
Awad Hegazy AbdelmonemCorresponding author Department of Human Anatomy and Embryology, Faculty of Medicine, Zagazig University, Zagazig 44519, Egypt

Background Hypothyroidism is a metabolic disorder affecting the functions of many tissues in the body including the testis. Testis is rich in the polyunsaturated fatty acids content and lacks strong intrinsic antioxidant system making it prone to such oxidative stress. L-carnitine (LC) regulates long chain fatty acids metabolism; and is considered a valuable antioxidant factor. Aim It was to evaluate the effect of hypothyroidism induced by propylthiouracil (PTU) on rats’ testes and the possible protective role of LC. Methods Forty-eight adult male albino rats were used in this work. The animals were divided into three groups with sixteen animals in each. Group 1 (Control): Animals were kept without medications. Group 2 (PTU-treated): was subjected to administration of PTU; while group 3 (PTU and LC) received both PTU and LC. By the end of the experiment “30 days”, blood samples were taken for hormonal assay; then animals were anaesthetized and sacrificed. Specimens were homogenized for biochemical analysis; epididymal content of each rat was obtained immediately for semen analysis. Testes’ specimens were harvested, prepared and examined by light microscope examination. Results Induced hypothyroidism was noticed to cause histopathological, morphometric and biochemical changes in rat’s testes. LC protected the testicular specimens against such changes; it also improved the seminal quality and quantity as well as testicular structure and biochemistry. Conclusion Hypothyroidism could result in hazards to the structure of testis. Fortunately co-administration of LC might reduce such hazards.

Evaluation of Anti-Aging Activity of the Biofield Energy Treated Novel Test Formulation Using SIRT1 and Telomerase Activity in in Vitro Model

Sep 2019 DOI 10.14302/issn.2474-7785.jarh-19-2994
Jana SnehasisCorresponding author Trivedi Science Research Laboratory Pvt. Ltd., Thane (W), India

Telomerase and SIRT1 (member of the sirtuin protein family) along with the lifestyle and diet are the major determinants of aging and its associated diseases such as cancer and cardiovascular disorders. The study objective was to investigate the effect of Consciousness Energy Healing based novel test formulation in pre-adipocytes (3T3-L1) and human peripheral blood mononuclear cells (PBMCs) for anti-aging activity using SIRT1 and telomerase assay. The test formulation was divided into two parts. One portion was denoted as the untreated test item without any Biofield Energy Treatment, while the other portion was defined as the Biofield Energy Healing Treatment, which received the Biofield Energy Healing Treatment by a renowned Biofield Energy Healer, Mahendra Kumar Trivedi. The cell viability using MTT assay showed that the cell viability of 3T3-L1 and PBMCs cells was more than 70% indicating a safe and nontoxic profile. The experimental data in PBMCs cells showed that the Biofield Energy Treated Test formulation showed a significant improved telomerase activity by 39.25%, 20.86%, and 17.95% at concentrations 0.01, 5, and 100 µg/mL, respectively as compared with the untreated test formulation group. These results indicate that the Biofield Energy Healing Treatment would be the significant approach to prevent aging-related disorders such as decline cardiovascular diseases, osteoporosis, dementia, osteoarthritis, Alzheimer’s, hypertension, cancer, Parkinson's Disease, Chronic Obstructive Pulmonary Disease (COPD), Stress, Asthma, cataract, age-related macular degeneration (AMD), hearing loss and metabolic disorders.

Impact of Biofield Energy Treatment Based Test Formulation on Vital Organ Health Specific Biomarkers Using Cell Line Study

Jul 2019 DOI 10.14302/issn.2640-6403.jtrr-19-2946
Jana SnehasisCorresponding author Trivedi Science Research Laboratory Pvt. Ltd., Thane (W), India

Multiple organ dysfunction syndrome or failure is one of the major concerns against healthcare services in order to maintain the normal function. The present study aimed to explore the impact of the Biofield Energy Treated test formulation on the function of vital organs such as bones, heart, liver, lungs, and brain using standard activity parameters in specific cell-based assays. The test formulation and cells medium was divided into two parts, one untreated (UT) and other part received the Biofield Energy Treatment remotely by a renowned Biofield Energy Healer, Ariadne Esmene Afaganis, Canada and was labeled as the Biofield Treated (BT) test formulation/media. The test formulation was tested for cell viability, and the data suggested that the test formulation was found safe and non-toxic against all the cell lines. Cytoprotective activity among the experimental groups showed a significant improved activity by 94.4% at 1 µg/mL in untreated medium (UT-Med) + Biofield Treated Test Item (BT-TI) group in human cardiac fibroblasts cells (HCF) cells, while 84.4% at 10 µg/mL in BT-Med + BT-TI groups in human hepatoma cells (HepG2), and 124% increased cytoprotective action at 1 µg/mL in UT-Med + BT-TI group in adenocarcinomic human alveolar basal epithelial cells (A549) cells as compared with the untreated test group. ALP activity in MG-63 cells was significantly increased by 85.9% at 10 µg/mL in the UT-Med + BT-TI group, while in Ishikawa cells showed maximum increased ALP activity by 59.2% at 0.1 µg/mL in BT-Med + BT-TI groups as compared to the untreated group. The percent protection of HCF (heart) cells (decreased of LDH activity) was significantly increased by 53% and 40.5% at 1 and 10 µg/mL concentrations respectively, in UT-Med + BT-TI group, while BT-Med + UT-TI group showed increased protection by 68.5%, 70.7%, and 16.8% at 0.1, 1, and 10 µg/mL respectively, and 86.5%, 62.5%, and 34.2% improved cellular protection at 0.1, 1, and 10 µg/mL respectively, in BT-Med + BT-TI group as compared to the untreated test group. The percent protection of HepG2 (liver) cells (decreased of ALT activity) was reported by 33.5%, 63.2%, and 99.2% at 10 µg/mL in the UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI groups, respectively compared to the untreated group. Cellular protection of A549 (lungs) cells (increased of SOD activity) in terms of percentage was increased by increased by 39.8% (at 10 µg/mL), 44% (at 25.5 µg/mL), and 59.7% (at 25.5 µg/mL) in the UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI groups, respectively compared to untreated group. Serotonin level was significantly increased by 59.2% (at 0.1 µg/mL), 190.3% (at 0.1 µg/mL), and 201% (at 1 µg/mL) in the UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI groups, respectively compared to untreated in human neuroblastoma cells (SH-SY5Y). However, the relative quantification (RQ) of vitamin D receptor (VDR) was significantly increased by 159.1% (at 50 µg/mL), 212.7% (at 1 µg/mL), and 278.3% (at 10 µg/mL) in the UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI groups, respectively as compared to the untreated in MG-63 cells. Thus, the present data concluded that the overall multiple organ health using various standard biomarkers in specific cell lines were significantly improved with respect to health of bones, heart, liver, lungs, and brain after treatment with the Biofield Energy treated test formulation (The Trivedi Effect®). Thus, it can be used as a complementary and alternative therapy approach against many multiple organ disorders such as coronary artery disease, arrhythmias, congenital heart disease, cardiomyopathy, cirrhosis, liver cancer, hemochromatosis, asthma, chronic bronchitis, cystic fibrosis, osteoporosis, etc.

Cell-Based Vital Organs Specific Biomarkers Assessment using Biofield Energy Based Novel Test Formulation

Jul 2019 DOI 10.14302/issn.2576-6694.jbbs-19-2944
Jana SnehasisCorresponding author Trivedi Science Research Laboratory Pvt. Ltd., Thane (W), India

The aim of the present study was to determine the impact of Biofield Energy Treated test formulation using six differentcell-lines. The test formulation/item (TI) and cell media (Med) was divided into two parts; one part was untreated (UT) and other part received Biofield Energy Treatment remotely by a renowned Biofield Energy Healer, Janice Patricia Kinney, USA and labeled as Biofield Energy Treated (BT) test item (TI)/media. Based on cell viability assay, test formulation was found as safe at tested concentrations. Cytoprotective activity of test formulation showed a significant restoration of cell viability by 60.6% (10 µg/mL), 67.5% (63.75 µg/mL), and 117.5% (63.75 µg/mL) in UT-Med + BT-TI, BT-Med + UT-TI, BT-Med + BT-TI, respectively compared to untreated in human cardiac fibroblasts cells (HCF) cells. Moreover, restoration of cell viability was improved by 64% and 127.3% in UT-Med + BT-TI and BT-Med + UT-TI, respectively at 1 µg/mL compared to untreated in human liver cancer (HepG2) cells. Cellular restoration in A549 cells was improved by 314% and 112.3% at 1 µg/mL in BT-Med + UT-TI and BT-Med + BT-TI, respectively than untreated. ALP activity in Ishikawa cells was significantly increased by 175.5%, 547.2%, and 220.8% in UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI, respectively at 0.1 µg/mL as compared to untreated. Additionally, in MG-63 cells showed increased ALP activity by 76.9%, 78.4%, and 79% in UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI, respectively at 50 µg/mL compared to untreated. The percent cellular protection of HCF (heart) cells (decreased of LDH activity) was significantly increased by 60.6% (10 µg/mL), 67.5% (63.75 µg/mL), and 117.5% (63.75 µg/mL) in UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI, respectively as compared to untreated. An improved HepG2 cells protection (represents decreased ALT activity) by 115.1% (1 µg/mL), 42.5% (25.5 µg/mL), and 60.8% (10 µg/mL) in UT-Med + BT-TI, BT-Med + UT-TI, BT-Med + BT-TI, respectively as compared to untreated. Percentage cellular protection of A549 (lungs) cells (represents increased of SOD activity) was significantly increased by 191.1% and 81.4% at 0.1 µg/mL in UT-Med + BT-TI and BT-Med + BT-TI, respectively as compared to untreated. Serotonin level was significantly increased by 31.8% (10 µg/mL) and 56.9% (25.5 µg/mL) in UT-Med + BT-TI and BT-Med + BT-TI, respectively compared to untreated in human neuroblastoma cells (SH-SY5Y). Relative quantification (RQ) of vitamin D receptor (VDR) was significantly increased by 304.3% (0.01 µg/mL), 128.4% (0.1 µg/mL), and 240% (0.1 µg/mL) in UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI, respectively compared to untreated in MG-63 cells. Thus, Biofield Energy Treated test formulation (The Trivedi Effect®) significantly improved organ specific functional biomarkers and would be useful for multiple organs health related to coronary artery disease, arrhythmias, congenital heart disease, cardiomyopathy, cirrhosis, liver cancer, hemochromatosis, asthma, chronic bronchitis, cystic fibrosis, osteoporosis, etc.

Structure Activity Relationship of Xanthones for Inhibition of Cyclin Dependent Kinase 4 from Mangosteen (Garcinia Mangostana L.)

Jun 2019 DOI 10.14302/issn.2379-7835.ijn-19-2845
J. Johnson JeremyCorresponding author University of Illinois at Chicago, College of Pharmacy, Department of Pharmacy Practice

The mangosteen fruit is a popular Southeast Asian fruit consumed for centuries. There have been a variety of xanthones isolated from the fruit, bark, roots and leaves with each having unique chemical and physical properties. Previously, the most abundant xanthone α-mangostin has been shown to inhibit CDK4. Herein we describe the role of selected xanthones from the mangosteen inhibiting CDK4. The evidence we provide here is that key functional groups are required to inhibit the CDK4 protein to prevent the phosphorylation of downstream targets critical to inhibiting uncontrolled cell cycle progression. To define the properties of xanthones for inhibiting CDK4 we utilized a cell free biochemical assay to identify inhibitors of CDK4. The following xanthones were used for the analysis: α-mangostin, β-mangostin, γ-mangostin, gartanin, 8-desoxygartanin, garcinone C and garcinone D, 9-hydroxycalabaxanthone, and 3-isomangostin These results further substantiate the unique pharmacological properties of individual xanthones and how a mixture of xanthones may be responsible for a multi-targeted effect in cell based pharmacology systems.

Behavioral Response and Acute Toxicity of Fingerlings of African Cat Fish, Clarias Gariepinus Exposed to Paraquat Dichloride

May 2019 DOI 10.14302/issn.2637-6075.jpae-19-2779
Chibueze Izah SylvesterCorresponding author Department of Biological Sciences, Bayelsa Medical University, Yenagoa, Bayelsa State, Nigeria.

This study evaluated the behavioural response and toxicity of paraquat dichloride to fingerlings of Clariasgariepinus. The fishes were acclimatized for 14 days and exposed to sublethal concentration of 0.00 ppm, 16.56 ppm, 22.08 ppm, 27.60 ppm, 33.12 ppm and 38.64 ppm. A 24 hours’ renewal bioassay was adopted in this study. Results showed that the fishes exhibited change in swimming, opercular movement, body pigmentation, surfacing and air gulping. Mortality rate increased significantly at p<0.05 as the concentration of the toxicant increased as well as the exposure period. LC50 values at 24, 48, 72 and 96 were 59.95, 47.59, 38.12 and 26.18ppm, respectively. Based on the results, Paraquat dichloride users need to discard the remains of empty cans properly to avoid contamination. Also there is need to exercise caution when using paraquat dichloride based herbicides in agricultural fields close to surface water resources.

Selective Cytotoxicity of Damsin Derivatives in Breast Cancer Cells

Apr 2019 DOI 10.14302/issn.2328-0182.japst-19-2759
Sterner OlovCorresponding author Centre for Analysis and Synthesis, Lund University, P.O.Box 124, 22100 Lund, Sweden

Cancer is the leading cause of death worldwide, and there is a constant need for new treatment strategies. Sesquiterpene lactones containing a 3-methylenedihydrofuran-2(3H)-one (or α-methylene-γ-lactone) moiety, for example damsin (1), are Michael acceptors that affect biological processes such as cell proliferation, death/apoptosis, and cell migration, by interfering with cell signalling pathways. Although the reactivity of the α-methylene-γ-lactone moiety is important for these effects, the Michael addition is reversible and it can be assumed that also other parts of the molecules will moderate any given biological activity. In this investigation, the cytotoxicity of 23 -methylene--lactones towards normal breast epithelial MCF-10A cells as well as breast cancer JIMT-1 cells is compared. Most of the investigated compounds are semisynthetic derivatives prepared by the condensation of the natural product damsin (1) with aldehydes. The two cell lines were treated with various concentrations of the compounds in dose response assays, and the 50 % inhibitory concentration (IC50) was determined from dose response curves. The IC50 values were found to depend strongly on the overall structure. The ratio between the IC50 values for MCF-10A and JIMT-1 cells, as a measure for the selectivity of a compound to kill cancer cells, was calculated, and found to vary between just over 1 to more than 10. The most potent derivatives formed from the condensation of 1 with aromatic aldehydes towards JIMT-1 cells are 3a and 3i, both with ratios between the IC50 values for MCF-10A and JIMT-1 cells close to 5. Also some aldol condensation products with acyclic aldehydes, i.e. 3r and 3u, were equally potent, and the latter showed the highest selectivity (ratio > 10). Structure-activity relationships that may explain the observed differences in potency and selectivity are discussed.

Antibacterial and Cytotoxicity Activities of Major Compounds from Tinospora cordifolia Willd. Growing on Mangifera indica L.

Apr 2019 DOI 10.14302/issn.2379-7835.ijn-19-2690
Taechowisan ThongchaiCorresponding author Department of Biology, Faculty of Science, Silpakorn University, Nakorn Pathom 73000, Thailand.

Objective To investigate the major constituents of Tinosporacordifolia Willd. growing on Mangiferaindica, and to evaluate the efficacy of their antibacterial and cytotoxicity activities. Methods The ethanolic stem extract of T. cordifolia was subjected to silica gel 60 column chromatography, thin layer chromatography and medium pressure liquid chromatography for isolation of the major compounds. Identification of purified compounds was achieved by spectroscopic methods.. The crude extract and purified compounds were screened for their antibacterial and cytotoxicity properties using standard procedures. Results Two alkaloids were purified and identified as Magnoflorin (1) and Tembetarine (2). These compounds showed high antibacterial activity against Bacillus cereus and Staphylococcus aureus with both MIC (32-64 µg/ml) and MBC (128-256 µg/ml). The cytotoxicity activity of the purified compounds and crude extract was determined using MTT colorimetric assay against L929 and HEK293 cell lines. This showed weak cytotoxicity activity with IC50 values of 1162.24 to 2290.00 µg/ml and 1376.67 to 2585.06 µg/ml towards L929 and HEK293 cell lines, respectively. Conclusion The major compounds present in ethanolic stem extract of T. cordifolia growing on M. indica were extracted, purified and identified. This study suggests that these compounds exhibit great potential for antibacterial activity with weak cytotoxicity activity. They may be useful for their medicinal functions.

Biolarvicidal Potentials of the Methanolic-Leaf-Extracts of Selected Tropical Plant Species

Apr 2019 DOI 10.14302/issn.2641-7669.ject-19-2730
A. Oyedeji AyodeleCorresponding author Department of Biological Sciences, Niger Delta University, Wilberforce Island, Nigeria

The global impact of malaria and challenges encountered during its control have necessitated the application of multifaceted strategies, including the application of plant-derived agents. Amidst these challenges the proliferation of the vector is becoming hyperendemic in tropical region. This research is focused on the biolarvicidal activities of the methanolic leaf-extracts of Cassia alata, Microdesmispuberula¸ and Spilanthesfilicaulis against mosquito larva. The mosquito larva were assayed in a static non-renewal test. Results showed no mortality for the negative control, and total mortality for the positive control (p<0.05). The C. alata bioassay was the most active with LC50 value of 13.73 ppm, followed by; M. puberula(21.24 ppm), and S. filicaulis (28.86 ppm). This study concludes that methanolic-leaf-extracts of C. alata, M. puberula¸ and S. filicaulis can be recommended for the formulation of biolarvicide for the control of malaria.

In vitro Assessment of the Biofield Treated Test Item on Cardiac Function Using Rat Cardiomyocytes Cell Line (H9c2) via Multiparametric Analysis

Feb 2019 DOI 10.14302/issn.2329-9487.jhc-19-2582
Jana SnehasisCorresponding author Trivedi Science Research Laboratory Pvt. Ltd., Thane (W), India

Introduction Heart disorders are the major concern of population health worldwide. According to WHO estimates 2018, 17.9 million peoples were died due to cardiovascular disorders. Aim The aim of this study was to investigate the cardioprotective activity of Biofield Energy Treated test item, Dulbecco's Modified Eagle Medium (DMEM) using rat cardiomyocytes (H9c2). Methods The test item (DMEM) was divided into three parts, first part received one-time Biofield Energy Treatment by a renowned Biofield Energy Healer, Mahendra Kumar Trivedi and was labeled as the one-time Biofield Energy Treated (BT-I) DMEM, while second part received the two-times Biofield Energy Treatment and is denoted as BT-II DMEM. The third part did not receive any treatment and defined as the untreated DMEM group. Results Cell viability of the test samples by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay showed 89.03% and 98.49% in the BT-I and BT-II groups, respectively suggested a nontoxic and safe in nature of the tested test item. The BT-I group showed 16.01% restoration of cell viability. The level of lactate dehydrogenase (LDH) was significantly inhibited by 50.37% and 49.35% in the BT-I and BT-II groups, respectively compared to the untreated DMEM group. Moreover, percent protection of creatine kinase-myocardial band (CK-MB) by 49.48% and 59.79% in the BT-I and BT-II groups, respectively, compared to the untreated DMEM group. Reactive oxygen species (ROS) level in terms of mean fluorescence unit (FU) was reduced by 6.64% in the BT-I group than untreated DMEM. Besides, BT-I and BT-II groups significantly increased the level of % apoptotic cells by 63.16% and 97.37% (p≤0.05), respectively than untreated DMEM. Conclusion Allover, results envisaged that Biofield Treatment significantly improved different cardiac parameters. Thus, Biofield Energy Treatment (The Trivedi Effect®) could be utilized as a cardio-protectant against several cardiac disorders such as coronary artery disease, heart attack, arrhythmias, heart failure, congenital heart disease, cardiomyopathy, etc.

Quantification of Micrornas by Absolute Dpcr for the Diagnostic Screening of Colon Cancer

Feb 2019 DOI 10.14302/issn.2471-7061.jcrc-18-2526
E. Ahmed FaridCorresponding author GEM Tox Labs, Institute for Research in Biotechnology, 2905 South Memorial Drive, Greenville, NC 27834, USA.

There is currently no validated micro(mi)RNA diagnostic stool test to screen for colon cancer (CC) on the market because of the complexity of fecal density, vulnerability of stool to daily changes, and the presence of three sources of miRNAs in stool (cell-free from fecal homogenates, exsosomal miRNAs from fecal exosomes, and fecal colonocytes). To address these complexities, we have first carried out a microarray miRNA experiment, using Affymetrix GeneChip miRNA 2.0 Arrays, on immunocaptured and enriched stool colonocytes of 15 subjects (three healthy controls and twelve colon cancer patients [three TNM stage 0-1 (e.g., polyps◻ ³ 1 cm, villous or tubvillous, or with high grade dysplasia), three stage 2, three stage 3, and three stage 4 in triplicates to select a smaller panel of 14 preferentially expressed mature miRNAs associated with colon cancer (12 Up-Regulated, miR-19a, miR-20a, miR-21, miR-31, miR-34a, miR-96, miR-106a, miR-133a, miR-135b, miR-206, miR-224 and miR-302; and 2 Down-Regulated, miR-143 and miR-145). In a subsequent validation study carried out on total small RNA extracted by immunocapture, followed by RT that employed TaqMan® miRNA Reverse Transcription (RT) Kit and a Custom TaqMan RT Primer Pool, absolute quantification of miRNAs, in copies/µl, was measured using a chip-based Absolute QuantStudio 3D Digital PCR analysis. To ensure that we have chosen human and not bacterial small total RNA, we have carried out coextraction protocols with E. coli K1 strain RS18, compare Agilent electrophoretic patterns, and also sequenced random samples throughout this research using mRNA/miRNA sequencing. Our initial quantitative dPCR miRNA data presented herein showe that the quantitative changes in the expression of a few mature miRNA genes in stool, which are associated with right and left colon cancer, would provide for a more convenient, sensitive and specific diagnostic screening markers thatare more useful than those test markers currently available on the market, such as the low-sensitivity (<15%) fecal occult blood test (FOBT); result in better compliance; and is more economical than the invasive and expensive colonoscopy exam in colon cancer, which can be cured if that cancer is detected at the early TNM stages, and that becomes incurable and deadly if not diagnosed before metastasis. Initial test performance characteristics of the miRNA approach showed that the test has a high numerical predictive value in colon cancer. Moreover, underpinning of the miRNA markers as a function of total RNA showed that the test can numerically differentiate between control subjects and colon cancer patients, particularly at the early stages of that curable cancer. We propose to extend our initial research results to a larger prospective and randomized five-years nested case-control study, to validate the expression of the above 14 miRNAs, in stool of 180 individuals in an epidemiologically designed study, using (30 controls and 150 colon cancer patients (thirty precancerous polyps (stage 0-1), forty five stage 2, and seventy-five colon cancer stages 3 or 4). chosen randomly by an epidemiological method from 900 control and CC subjects to allow for an adequate time to collect the required 900 stool samples, as well as allowing for statistically valid analysis, standardized test conditions, and to provide a mean for determining the true sensitivity and specificity of a miRNA-screening approach in noninvasive human stool. Power-analysis has indicated that a total of 180 individuals, which will take us 5 years to enroll in testing, is an appropriate number of subjects to standardize and validate our proposed miRNA screening test. We may find out at the end of the proposed validation study in stool that fewer miRNAs, or even one miRNA, may suffice to serve as an efficient and a quantitative marker for the non-invasive diagnostic screening of colon cancer in human stool. The above approach when combined with bioinformatics analysis, to correlate miRNA seed data with our previously published messenger (m)RNA target data in stool, allows for a thorough mechanistic understanding of how miRNA genes regulate mRNA expression, and would offer a better comprehensive diagnostic screening test for the non-invasive early detection stage (0-1) of colon cancer. In order to show the clinical sensitivity and specificity of the proposed miRNA test, the absolute miRNA PCR values, in copies/µl, will be correlated with FOBT, colonoscopy, and pathology data. Standardization will establish test’s performance characteristics (sample selection, optimal sample running conditions, preservation and storage) to ensure that the assay will perform the same way in any laboratory, by any trained personnel, anywhere in the World. Ultimately, a smaller number of selected validated miRNAs (<10) showing increased and reduced expression could suffice to give quantitative miRNAs colon cancer expression values, useful for the early diagnostic screening of that curable cancer.

Obesity Management Open Access

Beneficial Effects of Fennel (Foeniculum Vulgare) in Treating Obesity in Rats

Jan 2019 DOI 10.14302/issn.2574-450X.jom-18-2484
H. Radwan EmanCorresponding author Faculty of Science, Damanhour University, Egypt.

Obesity is associated with a number of serious medical complications, which are often referred to as the “insulin resistance syndrome”. The aim of the present study was performed to investigate the possible interaction between a conventional drug used for management of cholesterol and traditional herbal remedies on the obesity. This was carried through out: through estimation of blood test; Estimation of serum tests; Determination of oxidative stress biomarkers and the antioxidant enzymes activities in the liver were assayed; Histopathological examination of the liver and kidney of adult male albino rats were done. In the present study, the serum levels of the total protein and albumin in the obesity group (7.1± 0.2) and (4.78 ± 0.19); respectively were significantly (p ≤ 0.05) more than those of the control group (6.5±0.1) and (3.95± 0.1).The administration of (fennel group) revealed significant (P<0.05) decrease in the serum levels of the albumin and total protein (4.38± 0.1) and (6.65± 0.2); respectively as compared to the obesity group (4.78 ± 0.19) and (7.1± 0.2(. The total cholesterol of the group(5) (fennel and ator) after two weeks from a high fat diet than treatment with fennel and Ator through six weeks equal 142.86±5.9, 100.4±8.68, 93.29±5.99, 87.1±11.28, 80.4±21.55, 78.1±6.7 and 77.1±6.87; respectively. The present study showed a significant (P<0.05) increase in the activities of ALT, AST and ALP in the obesity group which recorded as (60.5±11.45), (57.25±6.3) and (845.0±49.47); respectively as compared to the control group (28.25±1.7), (38.5±3.87) and (537.0±41.5); respectively. The fennel group caused significant decrease in the activities of these enzymes (41.0± 2.9), (42.25+3.2) and (717.75+48.6); respectively compared to the obesity group. Ator group showed a significant decrease in the activities of these enzymes (40.0±2.16), (42.5±3.1) and (679.25±41.16); respectively compared as obesity group. The activity of AlT, AST and ALP in the fennel and ator group (32.75±2.5), (40.5±2.38) and (601.25±17.5); respectively were near to the control group.

Assessmentof Hair Growth Treatment with the Consciousness Energy Healing Treated Williams Medium E Using Mouse Vibrissae Hair Follicle Organ Culture

Jan 2019 DOI 10.14302/issn.2471-2175.jdrt-18-2520
Jana SnehasisCorresponding author Trivedi Science Research Laboratory Pvt. Ltd., Thane (W), India

Hair is playing an interesting part in human for social and sexual communication. Loss of hair follicle leads to various skin disorders. For this consequence, the present study has investigated the potential of the Biofield Energy Healing (The Trivedi Effect®) Treated test item (William’s Medium E) on the vibrissae hair follicle organ culture cells for the assessment of hair cell growth and development in vitro. The test item was divided into two parts. One part was defined as the untreated test item, where no Biofield Energy Treatment provided, while the other part was defined as the Biofield Energy Treated test item, which received the Biofield Energy Healing Treatment by renowned Biofield Energy Healer, Mahendra Kumar Trivedi. The study parameters like bulb thickness and formation of telogen were assessed using cell-based assay with the help of UTHSCSA Image tool version 3. The experimental results showed that the untreated test item group showed 20.9% and 28.2% increased bulb thickness on day 5 and 7, respectively compared to the day 1, while did not produce telogen follicles upto day 7. Besides, the percentage of telogen follicle was found as 43%, 57%, and 71% on day 3, 5, and 7, respectively of the Biofield Energy Treated test item group compared to the day 1. The overall results demonstrated that the Biofield Energy Treatment has the potential for hair growth promotion as evident via increased the formation of telogen. Therefore, the Biofield Energy Healing (The Trivedi Effect®) Treatment might be useful as a hair growth promoter for various treatment of skin injuries and skin-related disorders like necrotizing fasciitis, actinic keratosis, sebaceous cysts, diaper rash, decubitus ulcer etc.  

Veterinary Healthcare Open Access

Effect of Coinfection by Fasciola hepatica and Mycobacterium bovis on Bovine Tuberculosis Immunodiagnosis in an Enzootic Area Hidalgo State, Mexico.

Dec 2018 DOI 10.14302/issn.2575-1212.jvhc-18-2487
Fernando Díaz-OteroCorresponding author Centro Nacional de Investigaciones Disciplinarias en Microbiología Animal, Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias. Carretera México-Toluca, Km. 15.5, C.P. 05110, México D.F.

Parasitic infection by the Fasciola hepatica (F. hepatica) promotes susceptibility towards other infections, such as Mycobacterium bovis. As consequence, could affect diagnostic tests for this disease. Hence, the objective of this study was to assess the impact of F. hepatica coinfection on the most commonly used immunodiagnostic bovine tuberculosis (bTB) tests in field conditions in an enzootic area for both diseases. Thus, from a dairy herd located in Hidalgo State, México, displaying a 59.2% and 28% prevalence of fascioliasis and bTB, respectively. Sixty-one cows were analyzed based on their response towards bTB immunodiagnostic tests, such as Single Intradermal Comparative Tuberculin Test (SICTT), gamma-interferon test (BOVIGAM) and enzyme-linked immunosorbent assay (ELISA), along with the assessment of the F. hepatica parasite load and serodiagnosis by ELISA. Three study groups were formed according to test results. Group 1: coinfected (n=22). Group 2: non-parasitized cows, and positive for bTB tests (n=13) and Group 3: parasitized cows without tuberculosis (n=26). In addition, a group of cows kept in fascioliasis - and tuberculosis-free zones were included (Group 4, n=10). A non-parametric Kruskal-Wallis test and a Dunn test were applied to analyze the results. In Group 1, significant differences were observed regarding IFN-γ production, but not for antibody levels to M. bovis or reactivity towards bovine PPD in relation Group 2. While, Groups 1 and 3 did not display difference in antibody levels against F. hepatica. Differences were observed regarding tuberculosis and Fasciola diagnostic tests when both coinfected and infected groups were compared to controls. It is concluded that F. hepatica coinfection in tuberculous animals studied, depressed the production of IFN-γ towards bovine PPD under in vitro conditions, but its reactivity to the SICTT not show to be altered.

In Vitro Assessment of Estrogenic Potential of Biofield Energy Treatment using Human Endometrial Adenocarcinoma Cell Line

Dec 2018 DOI 10.14302/issn.2381-862X.jwrh-18-2459
Jana SnehasisCorresponding author Trivedi Science Research Laboratory Pvt. Ltd.,Bhopal, India

The objective of the study was to investigate the effect of Consciousness Energy Healing based DMEM medium on the level of alkaline phosphatase enzyme (ALP) activity in Ishikawa cells. The test item, DMEM medium was divided into two parts. One part of the test item received Consciousness Energy Healing Treatment by a renowned Biofield Energy Healer, Mahendra Kumar Trivedi and was labeled as the Biofield Energy Treated DMEM, while the other part did not receive any treatment, and defined as the untreated DMEM group. The cell viability using MTT assay of the Biofield Energy Treated DMEM group was observed as 108%, which indicated that the test item was safe and non-toxic. The estrogenic potential using ALP level showed a significantly increase by 73.21% in the Biofield Energy Treated DMEM group as compared to the untreated DMEM group. Overall, the experimental data suggested that the Biofield Energy Treated DMEM has significantly improved ALP level, which play a vital role for the promotion and maintenance of estrogen level. Based on the study outcomes, it is concluded that Biofield Energy Healing Treatment showed a significant improved ALP level, which can be used in various estrogenic disorders such as hypophosphatasia, osteoporosis, severe anemia, malnutrition, hypothyroidism, magnesium deficiency, heart surgery, aplastic anemia, chronic myelogenous leukemia, enteritis in children, Wilson’s disease, pernicious anemia, bacterial infection and intrauterine infection is a leading cause of pelvic inflammatory disease, subfertility, infertility, endometritis, early pregnancy loss, fetal defects, and preterm birth.

Antioxidant Activity Open Access

Antioxidative Potential of Consciousness Energy Healing Treatment on HepG2 Cells and DMEM after Oxidative Stress Induced by Hydrogen Peroxide

Oct 2018 DOI 10.14302/issn.2471-2140.jaa-18-2400
Jana SnehasisCorresponding author Trivedi Science Research Laboratory Pvt. Ltd.,Bhopal, India

Antioxidants can reduce oxidative stress in cells is used for the treatment of several disorders such as cancer, cardiovascular, and inflammatory diseases. The present study was evaluated the antioxidant potential of the Consciousness Energy Healing (The Trivedi Effect®) Treated human hepatoma cell line (HepG2) and Dulbecco's Modified Eagle Medium (DMEM) for the assessment of cell viability under hydrogen peroxide-induced oxidative stress. The Biofield Energy Treated HepG2 cells group was maintained for 23 days under standard conditions. On the next day, the cells were challenged with 1 mM of H2O2 for the generation of oxidative stress. The ability of the Biofield Energy Healing Treatment to protect from the oxidative stress was determined by MTT cell viability assay and compared with the negative control group. The percentage of cell viability was significantly (p≤0.001) increased by 13.6% in the Biofield Energy Treated DMEM group; while altered by 3.2% in the Biofield Energy Treated HepG2 cells group compared to the negative control groussp. Overall, the Biofield Energy Treated DMEM showed a better antioxidative protection against oxidative stress than HepG2 cells group, which was induced by H2O2. Therefore, the results envisaged that The Trivedi Effect®- Biofield Energy Healing Treatment has an impact on the protection of various vital organs from oxidative stress; which might be helpful in the development of powerful/energized growth medium for the accelerated study with a cost-effective manner.

Clinical Evaluation of Significance of 25(Oh)D Levels in Patients with Organophosphorus Poisoning

Oct 2018 DOI 10.14302/issn.2379-7835.ijn-18-2228
Kishan Gupta BalCorresponding author Senior Professor, In-charge Medical ICU, Department. of Medicine, S.P. Medical College, Bikaner.

Introduction: Organophosphate (OP) pesticide poisoning is a major challenging public-health problem in developing countries. Vitamin D deficiency is pandemic, yet it is the most under-diagnosed and under-treated nutritional deficiency in the world and it has been reported to be clinically correlated with psychiatric illness and manifestation of severe systemic inflammatory response syndrome like ARDS. Thus vitamin D deficiency may affect clinical course and outcome in cases of OPP. Aim: To evaluate status of 25 hydroxyvitamin D (25(OH)D) level in OP poisoning and its correlation with outcome of such patients. Materials and Methods: Serum 25(OH)D levels were measured at the time of hospitalization by electro-chemiluminescent Assay in 96 patients (76 male and 20 female) suffering from OP poisoning. Diagnosis of OP poisoning was made by history of poisoning including container of the poison brought by patient’s relative, clinical examination and measurement of serum butyrylcholinesterase activity. All patients were evaluated as per Performa and follow up till discharge. Results: Mean level of 25(OH)D in our cases was 24.57±9.91ng/ml and 66.7% had low levels of 25(OH)D. Our study shows linear relationship between 25(OH)D level and duration of hospital stay. All cases of OP poisoning who developed severe manifestations like ARDS, Intermediate syndrome (IMS) were having significant 25(OH)D deficiency. Our study also shows lower levels of 25(OH)D were associated with poor outcome (11.27±3.21vs 27.02±8.54, p<0.001). Conclusion: Vitamin D deficiency in OP poisoning is associated with longer hospital stay, more requirement of ventilator support and high prevalence of complication (ARDS and IMS) and poor outcome. Awareness of 25(OH)D level in patients with OP poisoning may be important to improve outcome.

Drug Design Progress of In silico, In vitro and In vivo Researches

Aug 2018
Bai QifengCorresponding author Key Lab of Preclinical Study for New Drugs of Gansu Province, School of Basic Medical Sciences, Lanzhou University, Lanzhou, Gansu 730000, P. R. China

Drug design, referred to the fields of pharmacology, biotechnology and medicine, is in silico, in vitro and in vivo assay processes of finding new candidate medications based on the biological targets. The in silicoexperiments of drug discovery are involved in the macromolecular structure databases, small molecule databases, molecular docking, de novo drug design and molecular dynamics simulations. The in vitro experiments of drug discovery need evaluate the direct interaction information between ligands and targets as well as the function of ligands on signaling pathway in the cell. The in vivo experiments of drug discovery give the convincing evidence for preclinical trial at the physiological level. In this review, we outline the drug design components of databases, virtual screening tools, biochemical assays, cell-based system and animal models.

Silver Nanoparticles: Cytotoxic and Apoptotic Activity on HT-29 and A549 Cell Lines

Aug 2018 DOI 10.14302/issn.2377-2549.jndc-18-2116
Sarani MinaCorresponding author Zabol Medicinal Plants Research Center, Zabol University of Medical Sciences.

Silver nanoparticles (Ag-NPs) are versatile materials with a broad range of applications in various fields such as cancer therapy, drug delivery. In this work, cytotoxic and apoptotic activities of silver nanoparticles was evaluation against lung (A549) and colon (HT-29) cell lines. The cytotoxic activity of nanoparticles was performed by MTT assay, while their apoptotic activity was tested through TUNEL method. The results of MTT of A549 have illustrate that fifty percent of cells destruction in concentrations more than 250 µg/ml of Ag-NPs. Apoptotic results of nanoparticles have shown more than fifty percent of apoptosis on A549 cell line. HT-29 display full apoptosis at concentrations more than 500 µg/ml. It seems that synthesized Ag-NPs by using P. farcta extract can be candidate as anti-cancer agent in treatment many cancers through creating or discovering new drug forms

Systems Biology Open Access

Ovarian Cancer Identification Based on Feature Weighting for High-Throughput Mass Spectrometry Data

Mar 2018
Liu XiaopingCorresponding author  School of Mathematics and Statistics, Shandong University at Weihai, Weihai 264209, China

An important use of proteomics data from Mass Spectrometry (MS) is the classification of tumor types with respect to peptides in specific cancer types. It is highly critical to find an optimal set of markers among specific cancer peptides whose expression can be clinically utilized to build assays for the diagnosis or to track the progression of specific cancer types. A number of feature selection algorithms have been proposed to obtain the classification of MS data. In this article, we proposed an improved feature selection algorithm based on feature weighting. Relief algorithm can calculate the weight of different features according to the correlation between their characteristics and categories. F-score is a simple filter-based feature selection method by evaluating how two sets of real numbers discriminate from each other. The main goal of this paper is to introduce a new feature weighting selection algorithm combining score from f-value and weight from relief, which is more accurate when classifying high-resolution MALDI-TOF (matrix-assisted laser desorption and ionization time-of-flight) MS data. We have developed a four-step strategy for data processing based on: (1) Align the study sets by binning of raw MS data, (2) local maximum search(LMS) peak detection, (3) a new combination feature weighting selection algorithm and (4) support vector machines achieve a satisfactory performance of identifying cancer and the healthy. The best parameter set for LMS were achieved with control variable method, which achieve an average accuracy of 97.4167% (sd = 0.0146) and the best accuracy of 98.6111% in 1000 independent 10 -fold cross validations. 

β-Cell function in type 1 diabetes may not be as low as presumed

Feb 2018 DOI 10.14302/issn.3070-2313.jeh-17-1756
Tamer GoncaCorresponding author Division of Endocrinology and Metabolism, Department of Internal Medicine, Medeniyet University, Göztepe Training and Research Hospital, Istanbul.

Objective We aimed to evaluate β-cell function of type 1 diabetic patients (T1DP)s based on fasting and stimulated C-peptide levels. Material and methods Study included 135 T1DPs and 31 healthy subjects. Fasting C-peptide levels were measured in healthy subjects and T1DPs. The Mixed-meal tolerance test (MMTT) was performed in T1DPs. Fasting and stimulated (90 minute post MMTT) C-peptide levels were measured via electrochemiluminescence assay. Two categorizations were made according to fasting (the first categorization ) and at 90th minute MMTT (the second categorization) C-peptide levels. For the first categorization; the groups were classified as follows: patients with undetectable ≤0.1ng/mL(group1); minimal 0.1-0.8ng/mL(group2); and sustained ≥0.8ng/mL (group3) C-peptide levels. For the second categorization, groups were as follows: patients with undetectable ≤0.1 ng/mL(group1); minimal 0.1-0.8 ng/ml (group2); and sustained ≥0.8ng/mL (group3) in which C peptide levels were increased to ≥150% of fasting C-peptide levels at the 90th minute after MMTT. Results For the first category; 41.5%, 40% and 18.5% of T1DPs were in group1, group2 and group3, respectively. For the second category; 34.8%, 20.7% and 44.4% were in group1, group2 and group3, respectively. In first categorization 58.5% and in second categorization 65.1% of T1DPs had detectable C-peptide levels. 44,4%of the T1DPs had a response to MMTT with C-peptide levels ≥0.8 ng/mL which increased to ≥150% of fasting C-peptide level at the 90th minute after MMTT as it is seen in non-diabetics. Conclusion The present study suggests the presence of functioning β-cells in T1DPs and 44.4 % of T1DPs have a response to MMTT as seen in non-diabetics. (ClinicalTrials.gov number: NCT02199470.)

Substrate Stiffness Influences the Time Dependence of CTGF Protein Expression in Müller Cells

Jan 2018 DOI 10.14302/issn.2578-8590.ipj-17-1910
J Foster WilliamCorresponding author Department of Physics, University of Houston, Houston, TX, USA

Following ocular trauma and retinal detachment, gliotic changes in the retina may develop over the subsequent month, a process known as PVR (proliferative vitreoretinopathy). There have been no successful therapeutic interventions to inhibit PVR. The protein CTGF (Connective Tissue Growth Factor) has been associated with retinal PVR and other fibrotic diseases of the retina in clinical studies but the mechanistic link between different pathologies and retinal gliosis has not been determined. In addition, CTGF has been previously noted to be associated, in some cases, with YAP/TAZ (Yes-associated protein and Tafazzin protein complex), transcriptional regulatory proteins that change subcellular localization in response to mechanical cues, such as the stiffness of the underlying material. We have previously shown that the mRNA for CTGF is markedly (100-fold) upregulated in retinal Müller cells grown on soft substrates. In order to evaluate if the mechanism by which mechanotransduction modulating CTGF production in retinal Müller cells involves the YAP/TAZ complex, this study tests the influence of substrate stiffness on the time dependence of CTGF protein expression, as well as subcellular localization of YAP/TAZ using a conditionally-immortalized mouse retinal Müller cell line plated on laminin-coated, polyacrylamide substrates of varying elastic modulus. Changes were assayed using immunohistochemistry and ELISA (Enzyme-Linked ImmunoSorbent Assay). In retinal Müller cells, the relationship between elastic modulus and the pattern of CTGF protein expression was bimodal, with CTGF levels rising more rapidly for cells on hard substrates and more slowly for cells grown on soft substrates. In addition, nuclear localization of YAP/TAZ corresponded directly to the maximum CTGF expression.

Human Health Research Open Access

Essential Oils Antagonism Against Three Hygiene Significant Yeasts and Juice Spoilage by Saccharomyces Cerevisiae

Nov 2017 DOI 10.14302/issn.2576-9383.jhhr-17-1768
Yassin Ibrahim SaharCorresponding author Botany Department, Faculty of Women for Arts, Science and Education, Ain Shams University, Cairo, Egypt.

Antifungal antagonism of different fourteen plant essential oils was examined as natural agents against economic and hygienic effective three yeasts; the preservative efficacy of most potent anti-yeast essential oil in food sanitary was also tested. Study involved oils antifungal bioactivity screening against Saccharomycescerevisiae, Candida albicans,and Candida utilis. Study also included selection and invitro extraction of most bioactive oil, and evaluation of its antifungal minimal inhibitory concentrations (MICs). Control ofjuice spoilage by Saccharomycescerevisiae under the effect of in vitro oil extract different concentrations was also screened. Among the tested essential oils, apricot seed oil was the most bioactive anti-yeast agent. Two MIC values of apricot oil invitro extract, 12.5μgml-1 and 25μgml-1 were recorded. In juice samples, oil extract bioactivity increased gradually up to concentration 100μgml-1.Highest oil preservative ability was observed at oil concentration of and above 125μgml-1. Higher oil concentrations needed for juice preservation were found more than in vitro assay to give the same effect. Applying of apricot oil and some other plant essential oils could be used as an environmental safety mode in osmophilic food preservation and in Candidate diseases biocontrol.

Evidence for the Absence of La Crosse Virus, Rift Valley Fever Virus, and Bunyamwera Virus in Korean Domestic Pigs

Oct 2017 DOI 10.14302/issn.2641-9181.ijnr-17-1778
Chung Hee-ChunCorresponding author Department of Veterinary Medicine Virology Laboratory, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul, Korea.

A surveillance study reports no detection of targeted bunyaviruses in domestic pigs in Korea. Methods, sampling frame, and assay limits are described to contextualize interpretation and surveillance value.

Immunization Open Access

Advanced Virological And Clinicopathological Studies On Cattle Suffering From Foot And Mouth Disease Virus

Sep 2017 DOI 10.14302/issn.2577-137X.ji-17-1736
Ahmed Kamal SamiaCorresponding author Animal Health Research Institute, Dokki, Giza, Egypt

Some strains of Foot and mouth disease virus (FMDV) are endemic in Egypt. The present study was performed on cattle and buffaloes (ages: 3 months up to 1.5 years old, of years 2015 and 2016), which were suffering foot and mouth disease (FMD). Sera and tissues samples were tested by different techniques including serum and virus neutralization tests (SNT, VNT), virus isolation and identification by tissue culture methods, Enzyme linked immune-Sorbent Assays (ELISA); and by the pathological and hematology techniques. The results showed the predominance of FMDV serotype O with the presence of serotypes SAT2 and A. The results showed the pathologic picture of FMD was similar regardless its specific subtypes, as apparently the studied strains produces same pathological and hematological changes. Microscopic examination reveals severe hydropic degenerations and necrosis in most affected organs, accompanied by significant changes in blood parameters which indicate severity and direct effects of FMDV on the hematopoietic system. These findings indicates the mode of pathogenesis of FMD virus in its way to exhibits the characteristic symptoms of illness. However, the investigation showed the presence of FMDV type O, A and SAT2 in the studied areas of delta governorates. It is important to focus on producing of vaccines which have only these serotypes as solution to get rid of the endemic behavior of FMDV in delta of Egypt.

Veterinary Healthcare Open Access

Assessment of the Immune Response Induced in Neonatal Calves by Vaccination with Mycobacterium Bovis BCG Phipps Under Field Conditions

Aug 2017 DOI 10.14302/issn.2575-1212.jvhc-17-1662
Díaz Otero FernandoCorresponding author Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias, Centro Nacional de Investigación Disciplinaria en Microbiología Animal, Carretera México-Toluca, Km 15.5, D.F., 05110, México

Immunization of cattle with the bacillus Calmette-Guérin (BCG) vaccine, especially neonates, induces protection against Mycobacterium bovis (M. bovis) and has been proposed as a strategy for bovine tuberculosis (bTB) control. Therefore, the aim of this study was to evaluate the immune response induced under field conditions in neonatal calves vaccinated with BCG Phipps, a strain that has rarely been evaluated in the bovine population, using interferon (IFN)-γ and tuberculin tests, flow cytometry and enzyme-linked immunosorbent assay. Two groups (vaccinated and control) of 5 calves were monitored for 12 weeks, and increases in the in vitro IFN-γ production, the percentage of cluster of differentiation (CD)8+ T cells and the activation levels of CD4+ and CD8+ T cells were observed 3 to 4 weeks post-vaccination. Bovine purified protein derivative-specific IFN-γ production was increased about 4.8- and 5.5-fold in vaccinated animals compared to non-vaccinated animals 3 and 4 weeks post-vaccination respectively. CD8+ T cells of the vaccinated group were increased 1.6-, 1.5- and 1.6-fold at weeks 2, 3 and 4 respectively. Levels of activation were 1.7- and 1.9-fold higher for CD4+ T cells and 2.3- and 1.8-fold higher for CD8+ T cells in the vaccinated group at weeks 3 and 4 respectively in response to M. bovis antigens. However, no animals (vaccinated or control) showed positive results for the single intradermal comparative tuberculin test (SICTT). Therefore, our results indicate that vaccination with M. bovis BCG Phipps strain stimulated peripheral blood T cell activity and induced a cell-mediated immune response. In addition, vaccination did not interfere with the SICTT, as previously reported, which indicates that this vaccine could be successfully applied in bTB control campaigns.

Veterinary Healthcare Open Access

Natural Infection by H1-Like Influenza a Virus in South American Camelids from Argentina: Serological Evidences.

Aug 2017 DOI 10.14302/issn.2575-1212.jvhc-17-1661
Parreño VivianaCorresponding author National Council of Scientific and Technical Research - CONICET, Buenos Aires, Argentina.

Serum samples from wild and domestic South American Camelids (SAC) from Argentina, collected before (2008), during (2009) and after (2010) the 2009 H1N1 influenza pandemic were tested by hemagglutination-inhibition assay (HIA) to evaluate the seroprevalence of antibodies (Ab) against different subtypes of influenza A viruses: A(H1N1)pdm09, A/sw/Argentina/SIV/2009(H3N2) and A/eq/Argentina/97(H3N8). For A(H1N1), an ELISA using a recombinant H1-hemmaglutinin from a reference strain (HA0 PuertoRico/8/1934) was also conducted. Serum samples from Guanacos (126), vicugnas (21) and llamas (100) from Jujuy, Mendoza and Río Negro provinces were analyzed; no clinical signs of respiratory disease were detected, reason for which no nasal swabs were obtained. No seropositive reactors to H3N2 nor H3N8 variants were detected, nevertheless high incidence of Ab reactive to A(H1N1)pdm09 were found by HIA; results which were confirmed by ELISA. The Ab seropositive animals to H1-like IAV found in llamas from Jujuy, and Mendoza (2009) were 78% and 86% by HIA and ELISA, respectively. Thirty-seven samples taken over the three years from guanacos kept in captivity in Rio Negro showed 62% of seropositive animals, while wild guanacos from Mendoza sampled in 2010 showed 36% seropositive animals to H1-like IAV, by both techniques. Finally, wild vicugnas from Jujuy, sampled in 2008 showed 38% and 52% seropositive animals to H1-like IAV by HIA and ELISA, respectively. Our results could indicate the potential role of these species as a reservoir of this zoonotic viral agent of high impact in Public Health, and may suggest that SAC populations might have been infected with an influenza strain antigenically related to H1 IAV. . Surprisingly, for llama and guanaco populations sampled over time in Jujuy and Río Negro, respectively, the HIA and ELISA geometric mean Ab titers (GMT) for 2008 were significantly higher than the ones of 2010. In addition, HIA and ELISA Ab titers found in domestic llamas were significantly higher than those detected in wild vicugnas sampled during that year (2008) in Jujuy. New field campaigns are in progress to collect serum samples and nasal swabs in order to isolate and characterize the virus responsible for triggering H1 reactive Abs. These findings remark the need to better understand the dynamics and ecology of influenza A virus within Sacs populations.

Serum Vitamin D Level in Oral Lichen Planus Patients of North India- A Case-Control Study.

Jun 2017 DOI 10.14302/issn.2471-2175.jdrt-17-1481
Prakash Sasankoti Mohan RaviCorresponding author Oral Medicine and Radiology, Subharti Dental College, India.

Background: Studies have unveiled lower levels of serum vitamin D in autoimmune diseases and the role of vitamin D in inhibition of helper T cells proliferation, stimulation of regulatory T cells, diminishing of B-lymphocyte differentiation and inhibition of immunoglobulin secretion has been discussed in the literature. Oral lichen planus (OLP) is an autoimmune mucocutaneous disorder in which cell mediated immunity plays an important part and so it may also be related to serum vitamin D levels. Aim: To estimate serum vitamin D in oral lichen planus patients of North India, compare it to the controls and to evaluate various factors influencing vitamin D level in OLP cases such as sun exposure, gender, psychosocial factors, meteorological influences, religion and diet. Methods and Material: Venous blood samples of 102 clinically diagnosed oral lichen planus patients and 102 age and sex matched controls were obtained for the study and serum vitamin D levels were estimated using Vitek Immunodiagnostic Assay System, an automated bench-top immuno-analyzer, based on the Enzyme Linked Fluorescent Assay. The data was analyzed using chi-squared test and t-test. Results: Mean vitamin D in OLP cases and controls was 20.40 ng/ml and 32.67 ng/ml respectively. Vitamin D3 deficiency was more in OLP cases (70.6%) as compared to controls (34.3%) and insufficiency was more in controls (35.3%) as compared to OLP cases (16.7%). These differences were statistically significant. Conclusion: Although vitamin D deficiency was found more in OLP cases pointing towards the possible co-relation of vitamin D and OLP, yet insufficiency was seen more commonly in controls which revealed that this important vitamin was lacking in North Indian population. 

In Vitro Cytoprotection of Resveratrol against H2O2-Induced Oxidative Stress and Injury in Astrocytes

Aug 2016 DOI 10.14302/issn.2474-9273.jbtm-16-1151
Xing GuoqiangCorresponding author Departments of Anesthesiology, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814

Oxidative stress mediated neural cell death is thought to be involved in the progression of secondary cell injury following brain trauma. Agents that can block oxidative stress-related injury could be potential therapies for TBI. Resveratrol, a polyphenol found in plants and red wine, is cytoprotective due to its potent antioxidant activities. To further understand how resveratrol could affect oxidative stress-induced injury, we hypothesized that the cytoprotective activities of resveratrol could be dose-dependent. In this study, resveratrol-induced cytoprotection was evaluated in cultured astrocytes. Primary rat astrocytes were cultured in T-75 flasks to a confluence of 80% before being plated onto 96-well plates. After 24 hours of acclimation, astrocytes were treated with various doses of hydrogen peroxide (H2O2) (0.1, 0.25, 0.5 and 1 µM) and resveratrol (25, 50, 75, 100 µM), respectively. Cell viability was determined 24 hours later using Alamar Blue Assay. Treatment of astrocytes with 0.5 mM H2O2, left 65% of astrocytes non-viable whereas treatment of astrocytes with 0.1 mM H2O2 had no effect on astrocytes viability; whereas 1 mM, H2O2 caused total loss of astrocyte viability. Resveratrol treatment at 75 µM and 100 µM has reduced 0.5 mM H2O2-induced cytotoxicity in astrocytes by 50%. Immunostaining with GFAP also confirmed these findings about the cytoprotective effects of resveratrol in astrocytes exposed to H2O2. These results suggest that resveratrol could be a potential neuroprotective agent in TBI due to its antioxidant properties. Further studies are needed to evaluate the long- term effects of resveratrol in animal models of TBI.

Metabolic Acidosis and Cardiovascular Disease in Patients on Peritoneal Dialysis

Jun 2016 DOI 10.14302/issn.2329-9487.jhc-15-905
D. Raikou VaiaCorresponding author Dpt of Medicine - Propaedaetic, National & Kapodistrian University of Athens, School of Medicine.

Backgroud: Metabolic acidosis, a common condition particularly in end stage renal disease patients, results in malnutrition and inflammation. In this study, we focused on the importance of metabolic acidosis on manifestations of cardiovascular disease in patients on peritoneal dialysis. Methods: We studied 20 patients on continuous ambulatory peritoneal dialysis (CAPD), 15 males and 5 females, on mean age 61.6 ±11.3 years old. Metabolic acidosis was determined by serum bicarbonate concentrations less than 22mmol/L, which were measured in gas machine. Dialysis adequacy was defined by total Kt/V/week for urea including peritoneal Kt/V for urea and residual GFR (ml/min/1.73m2). High sensitivity C-reactive protein (hsCRP) was measured using enzyme linked immunoabsorbed assay (ΕLISA). The concentrations of intact-parathormone (i-PTH) and beta2-microglobulin (beta2M) were measured by radioimmunoassays. Arterial stiffness was measured as carotid-femoral pulse wave velocity (c-f PWV) and augmentation index (AIx). We built a Cox regression analysis to predict coronary artery disease (CAD), congestive heart failure (CHF) and peripheral vascular disease (PVD). Results: Serum bicarbonate levels were inversely associated to beta2M, i-PTH and AIx (r=-0.451, p=0.04, r=-0.477, p=0.03 and r=-0.569, p=0.009 respectively). Cox- regression analysis revealed significant association of serum bicarbonate levels and PVD having as confounders traditional and specific for these patients risk factors. Conclusion: Metabolic acidosis may be an independent risk factor for arterial stiffening and peripheral vascular disease manifestation in patients on peritoneal dialysis.

Antioxidant Activity Open Access

New Antioxidant Flavonoids from the Aerial Parts of Secamone Afzelii

Feb 2016 DOI 10.14302/issn.2471-2140.jaa-15-887
Alabdul Magid AbdulmagidCorresponding author ICMR-UMR CNRS 7312, Groupe Isolement et Structure, Campus Sciences, Bât. 18, BP 1039, 51687 Reims Cedex 2, France

A bioassay-guided fractionation of petroleum ether, EtOAc and n-BuOH soluble parts of the 80% hydromethanol extract was performed to investigate the antioxidant activity of Secamoneafzeliiaerial parts using DPPH free radical scavenging assay. The results revealed that EtOAc and n-BuOH soluble parts have moderate to good DPPH radical scavenging activity (EC50 = 139.3 and 30.5 μg/mL, respectively). Therefore, from the most active fractions of EtOAc and n-BuOH soluble parts were isolated two new flavonoid diglycosides quercetin-3-O-β-d-apiofuranosyl-(1→2)-α-l-rhamnopyranoside and genkwanin-8-C-β-d-apiofuranosyl-(1→2)-β-d-glucopyranoside in addition to nine known compounds (2-10). Their structures were elucidated on the basis of spectroscopic data including 1D- and 2D-NMR and ESI-MS. The ability of the isolated compounds to scavenge the DPPH was evaluated. The new compound 1, quercitrin (3) and rutin (6) have antioxidant potential with EC50 values ranging from 8.4 to 13.6 µg/mL, compared to the standard ascorbic acid (EC50 7.4 µg/mL).

Antioxidant Activity Open Access

The Total Antioxidant Capacity of Foods: A Reappraisal. Application to Commercial Orange Juices

Dec 2015 DOI 10.14302/issn.2471-2140.jaa-15-715
Kevers ClaireCorresponding author Plant Molecular Biology and Biotechnology and CEDEVIT B22, University of Liège Chemin de la Vallée, 4, Sart Tilman, 4000 Liège Belgium

Since a few years, more and more attention has been specifically given to dietary antioxidants as agents promoting health and preventing the incidence of diseases. As part of these efforts, analytical methods and assays have been developed to measure the antioxidant content in food substances. In this paper, the antioxidant capacity of 17 orange juices is determined by various assays (DPPH, ORAC, heamolysis, xanthine/xanthine oxidase) as the content in ascorbic acid and total phenolics. The results evidence all the complexity to evaluate the in vitro antioxidant capacity of foods. In very general terms, in spite of the wide utilization in these tests (FRAP, TAC, ORAC TRAP and others), their significance remains obscure. The discrepancy of the results and the absence of good correlation between the assays clearly highlight all the importance of understanding the strengths and weakness of assays evaluating antioxidant potential of a food at the risk of giving erroneous information to the consumer. It is clear that the use of "total antioxidant capacity" assays for the in vitro assessment of antioxidant quality of food does not be employed by food industrials as a marketing argument or for the assessment of the "wholesomeness" of a food.

Antioxidant Activity Open Access

Radical-Scavenging and Anti-Oxidative Activities of TBN in Cell-Free System and Murine H9c2 Cardiomyoblast Cells

Dec 2015 DOI 10.14302/issn.2471-2140.jaa-15-765
Zhang ZaijunCorresponding author Institute of New Drug Research and Guangzhou Key Laboratory of Innovative Chemical Drug Research in Cardio-cerebrovascular Diseases, Jinan University College of Pharmacy, Guangzhou, 510632, China.

Reactive oxygen species (ROS) and reactive nitrogen species are believed to be one of the most important culprits in the pathogenesis of cardio/cerebrovascular diseases. Intensive researches have been conducted to target free radicals as potential treatment for cardio/cerebrovascular diseases. The 2-(((1,1-dimethylethyl) oxidoimino)-methyl)-3,5,6-trimethylpyrazine (TBN), a novel nitrone derivative of tetramethylpyrazine, has been demonstrated to exhibit significant therapeutic effects in ischemic stroke and Parkinson’s models due to its multiple functions, including calcium overload blockade and free radical-scavenging activity. In the present study, we found that TBN had significant radical trapping effect in cell-free assays. Additionally, TBN effectively blocked tert-butylhydroperoxide (t-BHP)-induced murine H9c2 cardiomyoblast cell death, suppressed H9c2 cell apoptosis and reversed the decrease in mitochondrial membrane potential. Furthermore, TBN markedly inhibited t-BHP-induced ROS generation and free radical NO and ONOO–.Taken together, these results suggest that TBN might be a potential candidate for the treatment of ischemic cardio/cerebrovascular diseases by targeting free radicals.

Antioxidant Activity Open Access

Comparison of Two Analytical Methods used for the Measurement of Total Antioxidant Status

Jun 2015 DOI 10.14302/issn.2471-2140.jaa-14-617
Srinivasa Rao P.V.L.N.Corresponding author Department of Biochemistry, Sri Venkateswara Institute of Medical Sciences, Tirupati, India.

Background: Antioxidants play an important role in maintenance of human health and prevention of disease. Effective supplementation of antioxidants requires laboratory monitoring of antioxidant status. An understanding of the methods used to determine the TAS helps in better interpretation of values obtained using a particular method and also to select a suitable method. Material and Methods: Forty subjects including 25 healthy volunteers and 15 patients diagnosed with rheumatoid arthritis were studied. All samples were analysed for TAS using Ferric reducing ability of plasma (FRAP) method and Trolox equivalent antioxidant capacity (TEAC) assay. Results: Mean TAS values obtained by TEAC method were higher than those obtained by FRAP method (p<0.0001); no difference was observed when TEAC values were corrected for proteins and FRAP values were corrected for uric acid (p=0.420). No correlation was found between TEAC and FRAP methods (p=0.102). However, when TEAC was corrected for proteins, positive correlation was observed with FRAP (p=0.044). There was agreement between the two methods when TEAC values were corrected for proteins. Conclusion: Although the reaction conditions differ, similar compounds react in both the assays and thus TEAC and FRAP assays are comparable. However, the two methods differ with respect to –SH groups and uric acid contributions. This contributes to the higher TAS values obtained by TEAC assay. Thus, in conditions with altered protein or uric acid levels, the two methods may not be used interchangeably. The TEAC assay is to be corrected for protein for comparison of reports of the two assays.

Influence of Chemical Refining Processes on the Total Phenolics and Antioxidant Activity of Sunflower Oil

May 2015 DOI 10.14302/issn.2379-7835.ijn-14-590
Jhaumeer Laulloo SCorresponding author Department of Chemistry, University of Mauritius

The raw sunflower oil (SFO) has an undesirable flavour and odour. Therefore, to make it suitable for human consumption, the oil has to undergo a number of refining processes such as degumming, neutralization, bleaching and deodorization. During these refining processes, some of the phytochemicals present in SFO is lost. The aim of this study is to evaluate the loss in total phenolic, flavonoid and non-flavonoid contents and the antioxidant properties of the oil at the different chemical refining stages. The crude SFO oil has the highest total phenolic, flavonoid and non-flavonoid contents. The amount of phenolic compounds decreases as the oil undergoes different chemical refining processes. Results of this study indicated that the highest percentage loss of the phenolic compounds occurred during the deodorizing step.The statistical loss of the deodorized oil was 41.7, 63.9 and 27.6 % for total phenolic, flavonoid and non-flavonoid contents respectively. The free scavenging activity of sunflower oil was determined using DPPH and ABTS assays. The percentage scavenging activity ranged from 55.64 to 35.87 % for the DPPH assay while for ABTS assay the activity ranged from 59.46 to 31.43 % in a 50 mg/ml of SFO sample. This showed a decrease in antioxidant activity from crude to DNW to bleached and deodorized oil. The crude oil having the highest phenolic contents showed the highest antioxidant activity in both DPPH and ABTS assays.

Acquired Abnormalities of Plasma Von Willebrand Factor Related Parameters and ADAMTS13 Autoantibodies in Aggressive Haematological Malignancies.

Jan 2015 DOI 10.14302/issn.2372-6601.jhor-14-377
Zaidah A WCorresponding author Department of Haematology, School of Medical Sciences, Universiti Sains Malaysia,Kubang Kerian, Malaysia.

Background Abnormalities of plasma von Willebrand Factor (vWF) system has been described in solid tumors but more information is required to understand the pathophysiological process in haematological malignancies. Objectives This study was carried out to investigate the changes in vWF-related parameters including ADAMTS13 protein level in aggressive haematological malignancies and to identify the prevalence of anti-ADAMTS13 antibody as well as its correlations with vWF-related parameters. Patients/Methods Patient newly diagnosed or having relapse acute leukaemias or aggresive non-Hodgkin lymphomas were recruited into this study. Exclusion criterias include; pregnancy, patient already commenced chemotherapy, sepsis or has background congenital bleeding disorders. Blood specimen was subjected to; blood counts, ADAMTS13 protein, ADAMTS13 antibody detection, vWF:Ag, vWF activity, factor VIII level (FVIII) and vWF: CBA (collagen binding assays) Results and Conclusion A total of 60 subjects with median age at 42.5 (IQR: 23.25-57.5) were included. There were 34(56.7%) lymphomas and 26(43%) acute leukaemias. FVIII, vWF:Ag, wVF activity and vWF:CBA level were elevated whereas ADAMTS13 protein was reduced in majority of patients. Those with lymphomas showed significantly higher levels of FVIII, vWF:Ag, vWF:activity and vWF:CBA compared to the leukaemias. 38(63.3%) of patients showed presence of ADAMTS 13 autoantibody. There was however no correlation between ADAMTS13 protein and vWF-related parameters or with ADAMTS13 autoantibodies. There was a high prevalence of ADAMTS 13 autoantibodies in this cohort despite the absence of thrombotic thrombocytopenic purpura (TTP). The more pronounced changes in vWF-related parameters among aggressive lymphomas compared to acute leukaemias are in tandem with the marginally higher rates of venous thromboembolism in the former.

Diagnostics of Progressive Multifocal Leukoencephalopathy in a Patient with Concomitant Lymphoma Infiltration of Central Nervous System During R-CHOP Chemotherapy- A Case Presentation and Review of the Literature.

Jun 2014 DOI 10.14302/issn.2372-6601.jhor-14-378
Kulma-Kreft MonikaCorresponding author Department of Radiotherapy and Medical Oncology, Gdynia Cancer Center, Gdynia

Progressive multifocal leukoencephalopathy (PML) is a rare complication associated, inter alia, with rituximab-based lymphoma treatment. PML diagnosis is made easier with the criteria recently published by the American Academy of Neurology. Unambiguous diagnosis of PML can be achieved by demonstration of the histopathological triad comprising:(1) demyelination, (2) bizarre astrocytes and (3) enlarged oligodendroglial nuclei together with detection of viral particles by electron microscopy. However, symptoms of PML may be similar to those observed during lymphoma progression into the central nervous system (CNS). Here we report the case of a patient with diffuse large B-cell lymphoma (DLBCL) treated with R-CHOP who developed clinical signs indicating PML. Intravital diagnostic methods failed to yield an unequivocal diagnosis of PML or lymphoma progression in the CNS. However, a post-mortem examination of brain biopsy specimens performed by electron microscopy demonstrated lesions typical for PML and the presence of viral particles. In addition, immunohistochemical assays identified a massive infiltration of lymphoma cells. The case thus suggests either the extremely rare coexistence of two complications: lymphoma CNS infiltration and PML or induction structural CNS lesions by lymphoma infiltration indistinguishable from PML. The presented findings thus highlight the need for a further review of the current diagnostic criteria for PML.

Development of Poly-ε-Caprolactone Based Nanoadjuvant for Effective Vaccination Against Tuberculosis

Apr 2014 DOI 10.14302/issn.2377-2549.jndc-13-329
Kumar Dinda AmitCorresponding author Department of Pathology, All India Institute of Medical Sciences,Ansari Nagar, New Delhi-110029 (INDIA)

The aim of the study was to synthesize sub-100nm poly-ε-caprolactone nanoparticles (PCL NP), load them with the mycobacterial protein, ESAT 6 and study the resulting immune responses in CD4+ and CD8+ T cells when incubated with human peripheral blood monocyte derived macrophages that had internalized the PCL NP. The synthesized PCL NP were characterized for size, shape and charge. They were found to be about 60nm in size with spherical shape. MTT assay revealed that the particles were perfectly biocompatible when tested in vitro on THP1 human monocytic cell line. The particles had a slow protein release kinetics and did not degrade appreciably even after 30 days in buffer solution. ELISA was used to quantify the cytokine response of CD4+ and CD8+ T cells when incubated with the monocyte derived macrophages as antigen presenting cells. The result of antigen presentation assay revealed that the antigen loaded PCL NP enhanced Th1 and CD8+ T cell responses significantly compared to the pure antigen. Thus we conclude that PCL NP of 60nm size can be effectively tested as a vaccine adjuvant with resulting activation of Th1/Th2 immunity as well as cytotoxic T cell response.

Phytochemicals May Arrest HIV-1 Progression

Jan 2014 DOI 10.14302/issn.2324-7339.jcrhap-13-edt.1.3
Sharma B.Corresponding author Department of Biochemistry, Faculty of Science,

This hypothesis‑driven review considers phytochemicals with putative anti‑HIV activity. It summarizes proposed mechanisms, in vitro evidence, and formulation issues, and calls for standardized assays and clinical trials to determine real‑world efficacy and safety.

Liquid Chromatography Tandem Mass Spectrometry Method for Determination of Febuxostat in Human Plasma to Support A Bioequivalence Study

Mar 2013 DOI 10.14302/issn.2328-0182.japst-12-173
N. Patel BhavinCorresponding author Bio-Analytical Laboratory, Cliantha Research Ltd., Bodakdev, Ahmedabad-380054, Gujarat, India.

A reliable, selective and sensitive liquid chromatography-tandem mass spectrometry (LC-ESI-MS/MS) assay has been proposed for the determination of febuxostat in human plasma using indomethacin as the internal standard (IS). The analyte and IS were extracted from 200 µL of human plasma via liquid-liquid extraction using methyl tert-butyl ether. Chromatography was performed on Hypurity C18 (100 mm × 4.6 mm, 5 µm) column under isocratic conditions. Detection of analyte and IS was done by tandem mass spectrometry, operating in negative ionization and multiple reaction monitoring mode. The deprotonated precursor to product ion transitions monitored for febuxostat and indomethacin were m/z 315.1 →271.0 and 356.1→312.0 respectively. The limit of detection (LOD) and limit of quantitation (LOQ) of the method were 0.0025 µg/mL and 0.05 µg/mL respectively. The linear dynamic range validated for febuxostat was 0.05-6.00 µg/mL. The intra-batch and inter-batch precision (% CV) was ≤ 7.1 % while the mean extraction recovery was > 87 % for febuxostat across quality control levels. The method was successfully applied to a bioequivalence study of 80 mg febuxostat tablet formulation in 14 healthy Indian male subjects under fasting and fed condition. The reproducibility in the measurement of study data was demonstrated by reanalysis of 110 incurred samples.

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