Standard Assay Research Articles

Purification of a chemotherapeutic agent, L-Arginase, from Pseudomonas aeruginosa isolated from soil.

L. arginase refers to the enzyme arginase found in the genus Lactobacillus, it plays a crucial role in the urea cycle, and has implications in various biological applications. This study aimed to purify arginase from Pseudomonas aeruginosa, isolated from soil, and apply it as an anticancer. 28 soil samples of P. aeruginosa were collected from different places of Baghdad, and rice lands in Najaf and Diwaniyah governorates. Different standard laboratory and biochemical assays, and Vitik system were used in diagnosis and growth of arginase enzyme under certain pH, temperature, incubation period. The purified enzyme was precipitated by ammonium sulfite (60-80%), dialyses bag 8000-1000KD, ion exchange by DEAE cellulose and sephadex G100 in gel filtration. Cytotoxicity of arginase against breast t cancer AJM-13 and rat embryo fibroblast REF normal cell line was evaluated for (48 and 72 hours). The inhibition rate increased in the low concentration of abnormal cell (AMJ-13) while decreased in the normal cell (REF), this study takes different concentration (0.392-12.5mg/mL), and low concentration (1562-0.048 mg/mL), the result in high concentration was IR 54.7% during 72 hours for AJM-13 and 14.3% for REF in the same time, while the low concentration was IR 91% in the 1562 mg/mL in the AMJ-13, and 51% in ERF, LD50 of arginase enzyme was 0.781 mg/mL that 41% during 72 hours for ERF, its save to normal cells. Arginase enzyme, at low concentrations, may have an inhibitory effect on cancer cells, and simultaneously, protect normal cell lines.

Assessment of Proximate, Vitamins, In Vitro Antioxidant, and Functional Properties of Some Polyherbal Formulations as Prospective Botanical Candidates for Pharmaceutical Applications

Study’s Novelty/Excerpt This study evaluates the combined effects of Turmeric, Coriander, and Mint (TCM) leaves on nutritional, functional, and antioxidant properties in polyherbal formulations, showing their superior glucose absorption capacity and antioxidant activity compared to single herbs. The research highlights a significant nutritional shift in the TCM formulation, providing evidence for its enhanced efficacy in nutraceutical applications. These findings show the potential of TCM as a novel botanical candidate for safe and effective pharmaceutical development. Full Abstract Conventional drugs have numerous side effects, toxicity, and microbial resistance, making them unsafe for consumption. Polyherbal formulations are popular due to their potency, low cost, clinical efficacy, safety, and success in chronic conditions. The study aims to evaluate the proximate, vitamin, functional, and antioxidant activity of polyherbal formulations of Turmeric, Coriander, and mint (TCM) leaves as potential botanical candidates for pharmaceutical screening. Thus, addressing these issues and improving access to drugs for individuals by focusing on polyherbal formulations (PHF) is a potential solution. All analyses conducted were determined according to standard assay guidelines. The proximate analysis shows no significant difference among the formulations (p > 0.05), with TCM showing a nutritional profile shift because of the combination of the herbs rather than single herbs. In the vitamin composition, Vitamin A displays a pattern among the formulations, with TC and CM showing higher values than TM and TCM, which are, in turn, better than the single herbs. Vitamin C fluctuates across the formulations. TM displays higher, while T shows the lowest. Vitamin E content remains relatively stable among the formulations with a difference from the single herbs. In the functional properties, GAC displays notable variations among the formulations. TCM exhibits the highest GAC at 14.7 %, indicating its superior ability to absorb glucose. TC, CM, TM, and the single herbs show a comparatively lower GAC value. WAC also varies among the formulations. TCM displays lower WAC compared to TC, TM and CM. The single herbs show the lowest WAC. OAC shows intriguing trends across the formulations. TCM and TC exhibit similar OAC, while CM shows a slightly higher OAC, and TM displays better than single herbs, showing no significant difference (p > 0.05). In the antioxidant Activity, TCM shows lightly higher scavenging activity in 40, 80 and 120 mg/mL concentrations than other formulations. The outcome of this study has led us to conclude that employing safe combinations of these herbal formulations for the production of nutraceuticals is recommended.

Effect of cross-platform gene-expression, computational methods on breast cancer subtyping in PALOMA-2 and PALLET studies

Intrinsic breast cancer molecular subtyping (IBCMS) provides significant prognostic information for patients with breast cancer and helps determine treatment. This study compared IBCMS methods on various gene-expression platforms in PALOMA-2 and PALLET trials. PALOMA-2 tumor samples were profiled using EdgeSeq and nanostring and subtyped with AIMS, PAM50, and research-use-only (ruo)Prosigna. PALLET tumor biopsies were profiled using mRNA sequencing and subtyped with AIMS and PAM50. In PALOMA-2 (n = 222), a 54% agreement was observed between results from AIMS and gold-standard ruoProsigna, with AIMS assigning 67% basal-like to HER2-enriched. In PALLET (n = 224), a 69% agreement was observed between results from PAM50 and AIMS. Different IBCMS methods may lead to different results and could misguide treatment selection; hence, a standardized clinical PAM50 assay and computational approach should be used.Trial number: NCT01740427

A Label-Free Optical Flow Cytometry Based-Method for Rapid Assay of Disinfectants' Bactericidal Activity.

Selecting the appropriate disinfectant to control and prevent healthcare-associated infections (HAIs) is a challenging task for environmental health experts due to the large number of available disinfectant products. This study aimed to develop a label-free flow cytometry (FCM) method for the rapid evaluation of bactericidal activity and to compare its efficacy with that of standard qualitative/quantitative suspension tests. The bactericidal efficiency of eight commercial disinfectants containing quaternary ammonium compounds (QACs) was evaluated against four strains recommended by EN 13727 (Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus hirae) and four multidrug-resistant pathogens. The proposed FCM protocol measures changes in scattered light and counts following disinfectant exposure, neutralization, and culture steps. Unlike other available FCM-based methods, this approach does not rely on autofluorescence measurements, impedance cytometry, or fluorescent dyes. The FCM scattered light signals revealed both decreased count rates and morphological changes after treatment with minimum inhibitory concentrations (MICs) and higher concentrations for all tested bacteria. The results from the FCM measurements showed excellent correlation with those from standard assays, providing a rapid tool for monitoring the susceptibility profile of clinical, multidrug-resistant pathogens to chemical disinfectants, which could support infection prevention and control procedures for healthcare environments. This label-free FCM protocol offers a novel and rapid tool for environmental health experts, aiding in the optimization of disinfectant selection for the prevention and control of HAIs.

Quantitative and equipment-free paper-based agglutination assay of bacterial cells.

Background: point-of-care (POC) tests are useful for bedside/home applications, emergencies, frequent follow-ups, and resource-limited areas. Limited quantitative and equipment-free POC assays have been reported. This study aims to develop, validate, and apply a simple, quantitative, paper-based POC assay. Methods: wax-channeled paper treated with specific anti-Brucella and anti-Salmonella antibodies was used for distance-based chromatographic elution of stained bacterial cell agglutinations. Results: a qualitative paper-based agglutination POC test was developed using color intensity, tail appearance, and "+/-" signs that clearly distinguish the positive and negative results. The optimization of the test for paper type, microfluidic channel design, antibody and bacterial cell concentrations, and elution methods was carried out. Quantitative assay transformation was successfully developed using the color intensity of the original reaction zone, intensity of elution tail, and distance-based migration that correspond to bacterial agglutination size. The migration distance of eluted bacterial agglutination bands corresponds to the target concentration with good linearity and minimal variability. Reporting of colored band migration with numbers using microfluidic patterns was used to enhance non-technical end-user applications. A distance-based POC assay prototype was then successfully used for the accurate detection of known and unknown samples in comparison with standard assays. Conclusions: the migration distance of an eluted stained bacterial agglutination correlated with anti-bacterial antibody concentrations. A simple, cheap, quantitative, and equipment-free paper-based POC assay of bacterial cell agglutination was developed. This test can be used for simple "+/-" results, thermometer-like quantification, or text reporting with numbers corresponding to target concentrations. The assay has extended applications to different human disease biomarkers.

An In Vitro Investigation of the Antiproliferative and Antimetastatic Effects of Levosimendan: Potential Drug Repurposing for Cervical Cancer.

Cervical cancer presents a significant challenge to the global health of women. Despite substantial advances in human papillomavirus (HPV)-related cervical cancer vaccines, non-HPV-related cervical cancer is still waiting novel therapeutic options. Drug repurposing has provided a promising approach to improve cancer therapy in recent years. Our study aimed to explore the potential in vitro antineoplastic effects of levosimendan on cervical cancer cells. The antiproliferative effects of levosimendan were investigated on cervical cancer cells using a standard MTT assay. Fluorescent double staining was performed to identify its ability to induce apoptosis and necrosis. The possible mechanism of action of levosimendan was explored using cell-cycle analysis. Furthermore, antimetastatic effects were investigated using a wound-healing assay and a Boyden chamber assay. Our results revealed that levosimendan exhibited the highest growth-inhibitory effect in the HPV-negative C33A cell line. However, the effects were modest compared to the standard agent, cisplatin. Cell-cycle analysis detected that levosimendan can induce cell-cycle arrest in C33A cells by increasing the G1 and G2/M phases, decreasing the S phase, and enhancing the hypodiploid subG1 population. Levosimendan inhibited cell migration and invasion in a concentration-dependent manner. As levosimendan showed antimetastatic efficacy, it could be considered for repurposing to contribute to overcoming resistance to therapy in cervical cancer.

Minimal interference of concizumab with standard clinical coagulation laboratory assays - An in vitro study.

Non-factor replacement therapies are emerging as prophylactic treatment options in haemophilia A or B (HA/HB) with and without inhibitors. Concizumab is an anti-tissue factor pathway inhibitor (TFPI) monoclonal antibody preventing factor (F)Xa inhibition and enhancing thrombin generation. Based on experience with other non-factor therapies and extended half-life products, there is a focus on potential interference with common clinical coagulation assays used to monitor patients treated with concizumab. To evaluate the impact of concizumab on standard clinical coagulation assays. Plasma samples (normal, HA/HB with/without inhibitors) in the presence/absence of added concizumab (250-16,000ng/mL) were analysed in clinical assays including activated partial thromboplastin time (aPTT), prothrombin time (PT), FVIII and FIX one-stage clot and chromogenic substrate assay, assays for detecting FVIII or FIX inhibitors and other assays for coagulation factors. Concizumab did not impact PT assays, but resulted in a small shortening of aPTT (up to 5 s in haemophilia plasma and 0.4 s in normal plasma). Concizumab had no, or only a minor impact on FVIII and FIX activity assays or Bethesda inhibitor assays. FXI and FXII activity in normal plasma, as measured by single factor aPTT-based assay, was significantly increased in the presence of concizumab (+11% each). This was also the case for FVII and FX measured by PT-based assays using plasma with 25% of FVII or FX (+64% and +22%, respectively). The presence of concizumab did not, or only slightly, influence the outcome of standard clinical coagulation assays relevant for HA and HB.

Real-time triplex loop-mediated isothermal amplification (LAMP) using a turbidimeter for detection of shrimp infectious hypodermal and hematopoietic necrosis virus (IHHNV).

The World Organization for Animal Health still regulates the infectious hypodermal and hematopoietic necrosis virus (IHHNV) in shrimp. The existing disease identification approach is time consuming, necessitates expensive equipment, and requires specialized expertise, thereby limiting the accessibility of shrimp disease screening on farms. Loop-mediated isothermal amplification (LAMP) is recognized for its ability to detect inhibitory substances with high sensitivity and specificity. We developed a real-time triplex LAMP assay that combines the simplicity of point-of-care testing with the accuracy of a turbidimeter. Using a set of three LAMP primers, our technology enables rapid DNA amplification in a single reaction within 45 min and with a low detection limit (10 copies/reaction). We tested 192 shrimp samples from different sources and demonstrated the clinical utility of our method, achieving 100% specificity (95% confidence interval = 93.40-100.00%), 100% sensitivity (97.36-100.00%), and 100% accuracy (98.10-100.00%) in detecting IHHNV DNA, with a high Cohen's kappa value (1) compared to the standard quantitative polymerase chain reaction assay. The high technology readiness level of our method makes it a versatile platform for any real-time LAMP assay, and its low cost and simplicity make it well suited for fast deployment and use in shrimp farming.

The biological and therapeutic assessment of a P(3HB-co-4HB)-bioactive glass-graphene composite biomaterial for tissue regeneration.

An ideal wound dressing should create a healing environment that relieves pain, protects against infections, maintains moisture, removes debris, and speeds up wound closure and repair. However, conventional options like gauze often fall short in fulfilling these requirements, especially for chronic or nonhealing wounds. Hence there is a critical need for inventive formulations that offer efficient, cost-effective, and eco-friendly alternatives. This study focuses on assessing the innovative formulation based on a microbial-derived copolymer known as poly(3-hydroxybutyrate-co-4-hydroxybutyrate), P(3HB-co-4HB) bioactive glass and graphene particles, and exploring their biological response in vitro and in vivo-to find the best combination that promotes cell adhesion and enhances wound healing. The formulation optimized at concentration of bioactive glass (1 w/w%) and graphene (0.01 w/w%) showed accelerated degradation and enhanced blood vessel formation. Meanwhile biocompatibility was evaluated using murine osteoblasts, human dermal fibroblasts, and standard cell culture assays, demonstrating no adverse effects after 7 days of culture and well-regulated inflammatory kinetics. Whole thickness skin defect using mice indicated the feasibility of the biocomposites for a faster wound closure and reduced inflammation. Overall, this biocomposite appears promising as an ideal wound dressing material and positively influencing wound healing rates.

Conquering the beyond Rule of Five Space with an Optimized High-Throughput Caco-2 Assay to Close Gaps in Absorption Prediction.

Current drug development tends towards complex chemical molecules, referred to as "beyond rule of five" (bRo5) compounds, which often exhibit challenging physicochemical properties. Measuring Caco-2 permeability of those compounds is difficult due to technical limitations, including poor recovery and detection sensitivity. We implemented a novel assay, with optimized incubation and analytics, to measure permeability close to equilibrium. In this setup an appropriate characterization of permeability for bRo5 compounds is achievable. This equilibrated Caco-2 assay was verified with respect to data validity, compound recovery, and in vitro to in vivo correlation for human absorption. Compared to a standard assay, it demonstrated comparable performance in predicting the human fraction absorbed (fa) for reference compounds. The equilibrated assay also successfully characterized the permeability of more than 90% of the compounds analyzed, the majority of which were bRo5 (68%). These compounds could not be measured using the standard assay. Permeability and efflux ratio (ER) were highly predictive for in vivo absorption for a large set of internal bRo5 compounds. Reference cut-offs enabled the correct classification of high, moderate, and low absorption. This optimized equilibrated Caco-2 assay closes the gap for a high-throughput cellular permeability method in the bRo5 chemical space.

Does the ultrasensitive HBsAg Next assay enhance Hepatitis B diagnosis? An evaluation of analytical performances

BackgroundAccurate laboratory confirmation for Hepatitis B diagnosis and monitoring are crucial. Recently an ultrasensitive immunoassay test, the HBsAg Next (HBsAgNx), has been reported approximately eight times more sensitive than current HBsAg assays. The aim of our study was to assess the analytical performances of this new test. Methodology253 clinical samples from Saint Louis University Hospital were analyzed, splitted into four panels: (1) routine prospectively screening serums (n = 196), (2) retrospective serum samples before HBV reactivation (HBV-R) (n = 18), (3) occult HBV infection (OBI) (n = 10) and (4) a selection of wild type HBV genotypes (n = 29) ResultsPanel 1, showed robust agreement with the HBsAg Qualitative II (HBsAgQII) assay (Cohen's kappa = 0.83). Despite this agreement, 7 false positive with the HBsAgQII assay were found negative with HBsAgNx. One OBI was detected only with HBsAgNx. Panel 2 showed potential time savings in diagnosing HBV-R using HBsAgNx among 4/18 HBsAg positives samples. Panel 3 highlighted the ability of HBsAgNx to detect HBsAg in OBI patients defined by negative for HBsAg with HBsAgQII assay and positive for HBV DNA. Furthermore, the HBsAgNx assay detected all different genotypes. ConclusionThe study highlights the effectiveness of the HBsAgNx assay, showing its performance. It excels in detecting weakly positive samples and addressing challenging cases. HBsAgNx assay demonstrates promising analytical performances, with improved sensitivity and specificity compared to standard HBsAgQII assay, able to detect all genotypes. Its potential impact on early detecting and monitoring reactivations, and occult infections could be very useful in clinical practice.

Microfragmented abdominal adipose tissue-derived stem cells from knee osteoarthritis patients aged 29-65 years demonstrate in vitro stemness and low levels of cellular senescence.

To investigate the level of cellular senescence in stem cells derived from microfragmented abdominal adipose tissue harvested from patients with knee osteoarthritis (OA). Stem cells harvested from microfragmented abdominal adipose tissue from 20 patients with knee OA, aged 29-65 years (mean = 49.8, SD = 9.58), were analysed as a function of patient age and compared with control cells exhibiting signs of cellular senescence. Steady-state mRNA levels of a panel of genes associated with senescence were measured by qPCR. Intracellular senescence-associated proteins p16 and p21, and senescence-associated β-galactosidase activity were measured by flow cytometry. Cellular proliferation was assessed using a 5-ethynyl-2'-deoxyuridine proliferation assay. Stemness was assessed by stem cell surface markers using flow cytometry and the capacity to undergo adipogenic and osteogenic differentiation in vitro. No correlation was found between cellular senescence levels of the microfragmented adipose tissue-derived stem cells and patient age for any of the standard assays used to quantify senescence. The level of cellular senescence was generally low across all senescence-associated assays compared to the positive senescence control. Stemness was verified for all samples. An increased capacity to undergo adipogenic differentiation was shown with increasing patient age (p = 0.02). No effect of patient age was found for osteogenic differentiation. Autologous microfragmented adipose tissue-derived stem cells may be used in clinical trials of knee OA of patients aged 29-65 years, at least until passage 4, as they show stemness potential and negligible senescence in vitro. Not applicable.

Sex-Related Differences in Oxidant and Antioxidant Profiles of Murine Kidney and Brain: A Focus on Sirt3-Mediated Regulation

Background: Sirt3 is a mitochondrial deacetylase with an important role in maintainance of cellular redox and metabolic homeostasis and mitochondrial function. As growing evidence support the existence of sex-specific responses to metabolic and oxidative stress, we aimed to investigate sex- and organ-specific effects of Sirt3 loss. Materials and methods: Expression of Sirt3, PGC-1a, CuZnSOD, MnSOD and Cat proteins in kidneys and brains of Sirt3-wild type (Sirt3 WT) and Sirt3-knockout (Sirt3 KO) mice was assessed by Western blotting. Protein carbonylation and lipid peroxidation levels were measured using ELISA and fluorometric assays, respectively. SOD and Cat activities were determined using standard enzymatic assays. Results: Significant sex- and organ- specific differences in response to Sirt3 loss were detected. Sirt3 knockout affected kidneys more than brain tissue, with females showing lower levels PC and LPO. In kidneys, female KO showed higher MnSOD, but lower CuZnSOD and Cat activity compared to males. In brains, WT females show higher activities of these enzymes than males, suggesting a sex-specific protection mechanism, but female KO brains show a larger decrease in these parameters. Conclusion: Our study provides comprehensive insights into the complex interplay of Sirt3, oxidative stress, and antioxidant defenses in murine kidney and brain. The observed differences between the two organs and the impact of sex highlight the need for studying Sirt3 function in diverse physiological contexts. The tissue-specific responses and sex-related variations underscore the importance of considering these factors in the development of therapeutic strategies targeting mitochondrial function and redox homeostasis.

Investigating the Cardio-protective Activity of Agaricus bisporus in Wistar Rats Induced with Myocardial Infarction

Myocardial infarction is now recognized as one of the chronic diseases and the leading cause of death across the globe. The use of plants abundant in our environment, especially Agaricus bisporusprovides a cheap alternative to combat this common foe of mankind. Hence, this study was carried out to evaluate the cardio-protective effect of the leaf extract of Agaricus bisporus in isoproterenol-induced myocardial infarction. Fresh sample of Agaricus bisporus were purchased from a market at Asaba, Delta State, Nigeria. The samples were thereafter shredded with a knife and oven-dried at 40oC and pulverized and extracted, using Solvent-solvent (ethanol and water) (7:3) via maceration for 72 hr. The filtrate was evaporated to dryness to obtain the ethanol extract which was used for further bioassay study. The animals were administered with the extract of A. bisporus orally for fourteen consecutive days at a divided dose of 100 mg/kg, 200 mg/kg and 400 mg/kg body weights and thereafter 0.2 ml isoproterenol (ISO) at 150 mg/kg was injected intraperitoneally at an interval of 24 h on the 15th and 16th day. Subsequently, the blood pressures were monitored and blood collected for bioassay studies. Assay of High density lipoprotein cholesterol (HDL-c), low density lipoprotein cholesterol (LDL-c), triglyceride (TG) and total cholesterol (TC) levels as well as activities of lactate dehydrogenase (LDH), were carried out with standard assay kit sourced from Randox laboratories Ltd, United Kingdom with maximum adherence to the manufacturer’s instruction. Results showed a significant increase (P<0.05) in high density lipoproteins (HDL) levels and lactate dehydrogenase (LDH) activities with a significant decrease (P>0.05) in low density lipoproteins (LDL) and triglyceride levels in a dose-dependent manner (100<200<400 mg/kg body weight) among the extract treated groups when compared to the untreated control group. Conversely, total cholesterol levels of extract treated groups showed no significant difference when compared with the untreated control group.The findings of this work have shown that extract of Agaricus bisporus could help to mitigate cardiovascular diseases and could be used to produce plant based products to combat myocardial infarction, thereby improving general wellbeing.

Label-Free Screening of Drug-Induced Liver Injury Using Stimulated Raman Scattering Microscopy and Spectral Phasor Analysis.

Hepatic toxicity is a leading cause of the termination of clinical trials and the withdrawal of therapeutics following regulatory approval. The detection of drug-induced liver injury (DILI) is therefore of importance to ensure patient safety and the effectiveness of novel small molecules and drugs. DILI encompasses drug-induced steatosis (DIS) and drug-induced phospholipidosis (DIPL) which involve the accumulation of excess intracellular lipids. Here, we develop hyperspectral stimulated Raman scattering (SRS) microscopy as a label-free methodology for discriminating DIS and DIPL in mammalian cell culture. We demonstrate that hyperspectral SRS imaging in tandem with spectral phasor analysis is capable of discriminating DIS and DIPL based on the nature and distribution of intracellular lipids resulting from each process. To demonstrate the practical application of this methodology, we develop a panel of alkyne-tagged propranolol analogues that display varying DILI effects. Using hyperspectral SRS imaging together with spectral phasor analysis, our label-free methodology corroborated the standard fluorescence-based assay for DILI. As a label-free screening method, it offers a convenient and expedient methodology for visualizing hepatotoxicity in cell cultures which could be integrated into the early stages of the drug development process for screening new chemical entities for DILI.

Biochemical Markers for Neuroendocrine Tumors: Traditional Circulating Markers and Recent Development—A Comprehensive Review

Neuroendocrine neoplasms (NENs) are a heterogeneous group of neoplasms presenting unique challenges in diagnosis and management. Traditional markers such as chromogranin A (CgA), pancreatic polypeptide (PP), and neuron-specific enolase (NSE) have limitations in terms of specificity and sensitivity. Specific circulating markers such as serotonin and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) and various gastrointestinal hormones such as gastrin, glucagon, somatostatin, and vasoactive intestinal peptide (VIP) have a role in identifying functional NENs. Recent advances in molecular and biochemical markers, also accounting for novel genomic and proteomic markers, have significantly improved the landscape for the diagnosis and monitoring of NENs. This review discusses these developments, focusing on both traditional markers such as CgA and NSE, as well as specific hormones like gastrin, insulin, somatostatin, glucagon, and VIP. Additionally, it covers emerging genomic and proteomic markers that are shaping current research. The clinical applicability of these markers is highlighted, and their role in improving diagnostic accuracy, predicting surgical outcomes, and monitoring response to treatment is demonstrated. The review also highlights the need for further research, including validation of these markers in larger studies, development of standardized assays, and integration with imaging techniques. The evolving field of biochemical markers holds promise for improving patient outcomes in the treatment of NENs, although challenges in standardization and validation remain.

Investigating the Cardio-protective Activity of Volvariella volvacea in Isoproterenol-induced Myocardial Infarction Using Animal Model: An In vivo Approach

Ischemic heart disease (IHD) is a severe cardiovascular condition characterized by impaired blood flow to the heart muscle, leading to tissue damage and dysfunction. Among different natural resource, mushrooms are now becoming more attractive because of its strong nutritional value and therapeutic potentiality. Wild edible fleshy fungi like Volvariella volvacea have been instrumental in preventing and treating chrome diseases. Hence, this study was carried out to evaluate the cardio-protective activity of Volvariella volvacea in isoproterenol-induced myocardial infarction using animal model. Twenty five (25) male wistar rats were randomly grouped into five classes (A, B, C, D and E) of five rats each. Groups A, B and C were pre-treated with 100mg/kg, 200mg/kg and 400mg/kg of Volvariella volvacea extract respectively. Group D was administered 10 mg of lisinopril (standard drug), this served as the positive control group while group E was administered 0.2 ml distilled water, and this served as the negative control group. All the rats were intraperitoneally induced with 150 mg of isoproterenol after fourteen days (two weeks) of pretreatment. Cardiac biomarker (lactate dehydrogenase (LDH)), was measured to assess cardiac injury, using standard method while lipid bioassay such as high density lipoprotein (HDL), low density lipoprotein (LDL), triglycerides (TG) and total cholesterol (TC) were assayed using standard assay kits. Results showed that rats pre-treated with Volvariella volvacea extract showed significant effect on cardiac biomarkers compared to the control group. Results showed a significant increase (P<0.05) in high density lipoproteins (HDL) levels and lactate dehydrogenase (LDH) activities with a significant decrease (P>0.05) in low density lipoproteins (LDL) and triglyceride levels in a dose-dependent manner (100<200<400 mg/kg body weight) among the extract treated groups when compared to the untreated control group. However, total cholesterol levels of extract treated groups showed no significant difference when compared with the untreated control group. This study has shown that extract of Volvariella volvacea could help to mitigate cardiovascular diseases and could be used to produce plant based products to combat Ischemic heart disease, and consequently improving general wellbeing.

Multiplexed colorimetric assay of antioxidants in wines with paper-based sensors fabricated by pen plotting

This work reports the development of low-cost and rapid multiplexed colorimetric assay of antioxidants (total phenolics, antioxidant capacity, flavonoids and anthocyanins) in wines at daisy-shaped fluidic paper-based analytical devices (PADs). The desired fluidic patterns were formed on paper by pen drawing and colorimetric reagents were immobilized at the 6 peripheral test zones. The sample was added at the central sample zone, migrated to the test zones and reacted with the immobilized reagents producing characteristic colors that were captured and analyzed. The paper-based approach was applied to the analysis of several wine samples and the results were statistically correlated to standard solution-based colorimetric assays, indicating that it could be reliably used for ranking wines according to their antioxidants content. In addition, the paper-based analytical methodology is simple, instrument-free, portable, cost-effective, rapid and environment friendly.

Hypovitaminosis D among type 2 diabetes patients and non-diabetics at Libreville

: Type 2 diabetes is resulting from insulin resistance and/or progressively reduced insulin secretion. Vitamin D nuclear receptor present on the β cells of the pancreas promotes the growth, synthesis and secretion of insulin. Furthermore, vitamin D deficiency has been associated with numerous disorders, such as type 2 diabetes, cardiovascular diseases, arterial hypertension, cancer, multiple sclerosis, depression, psychiatric diseases. Determine the frequency of hypovitaminosis D among type 2 diabetes patients and those without diabetes in Libreville. The study population was composed of 92 type 2 diabetes patients (cases) and 100 non-diabetics (controls). The vitamin D and glycated hemoglobin were performed using 25 OH Vitamin D total kit and FIA8000 kit respectively. The blood glucose, calcemia and phosphorus were performed using standard spectrophotometric assay. Insufficient vitamin D (<30ng/mL) was found among 55.2% of the study population that was 57.0% of insufficient vitamin D in cases and 53.3% in controls. In the general population vitamin D was not correlated to blood glucose (r= -0.0270). Thus, with glycated hemoglobin, no correlation was found with vitamin D among cases. However, a weak negative correlation of vitamin D with blood glucose has been found in control group (r= -0.2798). High hypovitaminosis D was found among cases and controls. Therefore and in regards of studies based on vitamin D supplementation, with an appropriate concentration of vitamin D or calcitriol, glucose homeostasis may be maintained. Moreover, implementation of preventive measures should be done to avoid disastrous consequences of vitamin D insufficiency.

Development and standardization of an enzyme-linked inmunosorbent for the detection of orthohantavirus infection in Argentina based on its bacterial-expressed nucleocapside protein.

We conducted a development and standardization of an IgG ELISA assay for serological detection of human orthohantavirus infections using the recombinant antigen rLECH13 produced in bacterial and derived from the LECHV. The evaluation and standardization were carried out by analyzing serum samples from a total of 50 patients with confirmed Hantavirus Pulmonary Syndrome (HPS) diagnosis through the reference technique, 50 negative sera, and 53 patients with other medical conditions. The data from the assay analysis showed a diagnostic sensitivity value of 95% and a diagnostic specificity of 80%. The high sensitivity of this novel assay leads us to conclude that rLECH13 is a feasible option for use in the immunodiagnostic of orthohantavirus infection. Additionally, it is crucial to have an antigen that can be produced under conditions that do not require highly complex laboratories. Furthermore, the new assay is cost-effective, reproducible, and demonstrates excellent performance.