Enzyme-linked Immune Sorbent Assay Research Articles

Sirtuin 3 protects against anesthesia/surgery-induced cognitive decline in aged mice by suppressing hippocampal neuroinflammation

BackgroundPostoperative cognitive dysfunction (POCD) is a very common complication that might increase the morbidity and mortality of elderly patients after surgery. However, the mechanism of POCD remains largely unknown. The NAD-dependent deacetylase protein Sirtuin 3 (SIRT3) is located in the mitochondria and regulates mitochondrial function. SIRT3 is the only sirtuin that specifically plays a role in extending lifespan in humans and is associated with neurodegenerative diseases. Therefore, the aim of this study was to evaluate the effect of SIRT3 on anesthesia/surgery-induced cognitive impairment in aged mice.MethodsSIRT3 expression levels were decreased after surgery. For the interventional study, an adeno-associated virus (AAV)-SIRT3 vector or an empty vector was microinjected into hippocampal CA1 region before anesthesia/surgery. Western blotting, immunofluorescence staining, and enzyme-linked immune-sorbent assay (ELISA) were used to measure the oxidative stress response and downstream microglial activation and proinflammatory cytokines, and Golgi staining and long-term potentiation (LTP) recording were applied to evaluate synaptic plasticity.ResultsOverexpression of SIRT3 in the CA1 region attenuated anesthesia/surgery-induced learning and memory dysfunction as well as synaptic plasticity dysfunction and the oxidative stress response (superoxide dismutase [SOD] and malondialdehyde [MDA]) in aged mice with POCD. In addition, microglia activation (ionized calcium binding adapter molecule 1 [Iba1]) and neuroinflammatory cytokine levels (tumor necrosis factor-alpha [TNF-α], interleukin [IL]-1β and IL-6) were regulated after anesthesia/surgery in a SIRT3-dependent manner.ConclusionThe results of the current study demonstrate that SIRT3 has a critical effect in the mechanism of POCD in aged mice by suppressing hippocampal neuroinflammation and reveal that SIRT3 may be a promising therapeutic and diagnostic target for POCD.

Faecal Calprotectin in Differentiating Inflammatory Bowel Disease (IBD) from Irritable Bowel Syndrome (IBS)

Inflammatory bowel disease (IBD) is a chronic idiopathic inflammatory disorder of the gastrointestinal tract with relapsing and remitting course. Irritable bowel syndrome (IBS) is a gastrointestinal disorder characterized by altered bowel habit in association with abdominal discomfort and pain. Faecal biomarker may be used an accurate tool in the differentiation of IBD and IBS. The aim of this study was to measure faecal calprotectin (FC) level in patients with IBD and IBS and compare between them. This cross-sectional observational study conducted at the department of Gastroenterology, BSMMU, Dhaka, Bangladesh. Patients with IBD were diagnosed on the basis of compatible history, clinical examination, laboratory, radiological and endoscopic findings, where IBS patients were selected by using the Rome IV criteria. Quantitative faecal calprotectin enzyme-linked immune sorbent assay (ELISA), BÜHLMANN Quantum Blue® test was done and compared between IBD and IBS patients. In this study, Ninety (90) patients were enrolled, 45 patients with IBD and 45 patients with IBS. Mean age of the IBD patients was 32.24±9.76 years and IBS patients was 33.80±9.70 years. There were 28 (62.2%) male and 17 (37.8%) female patients with IBD and 30 (66.7%) male and 15 (33.3%) female patients with IBS. We found faecal calprotectin ( FC) level was 445.68 ± 237.35µg/g in IBD patients and 39.16 ± 17.31µg/g in IBS patients. There was a significant difference of faecal calprotectin level between IBD and IBS patients (p-value < 0.001). The sensitivity and specificity of faecal calprotectin to differentiate IBD from IBS was 91.1% and 86.7% respectively. The test accuracy was 88.9%. Area under ROC was 0.959 (95% CI, 0.909 to 1.0). This study showed that faecal calprotectin appears to be clinically useful, non-invasive, rapid and reliable marker to differentiate IBD from IBS. Bangladesh Med J. 2021 January; 50(1) : 15-22

Circulating Endocan in Subjects with Obesity and Carbohydrate Disturbances

Objective: Endothelial-specific molecule 1 (endocan) is expressed by endothelial cells and may have a major role in the regulation of cell adhesion and in the pathogenesis of inflammatory disorders such as obesity and diabetes. Аim of this study is to evaluate endocan serum levels in patients with obesity and carbohydrate disturbances. Subjects and methods: The study group consisted of 163 patients with mean age 52,5±11,3 years, divided in four groups - obesity without carbohydrate disturbances, prediabetes, diabetes and 42 healthy individuals as a control group. Endocan levels were determined using a commercially available human enzyme-linked immune sorbent assay (ELISA) kit. Results: The levels of endocan were significantly higher in patients with prediabetes and obesity compared to the control group (618.92±874.29 ;425.85±391.76; vs.321.02±304.11pg/ml, p<0.05). Correlation analysis revealed that endocan correlates positively with triglyceride (TG) (r=0.219;p<0.05), gamma-glutamyl transferase (GGT) (r=0.225;p<0.05), visceral adiposity index (VAI) (r=0.186;p<0.05) and negative with HDL (r= -0.176;p<0.05). We found higher level of endocan in patients with three or more components of MS as well as higher levels of endocan were found in patients with dyslipidemia (p<0.05). ROC analyses determined circulating endocan levels to be of value for differentiating subjects with prediabetes (AUC=0.670,p=0.007, as a cut-off value of ≥183pg/ml had a sensitivity of 88% and specificity of 65%) Conclusion: The levels of endocan were increased in patients with obesity and carbohydrate disturbances. Further studies will elucidate the role of the biomarker in development of type 2 diabetes and its complications

Influence of Supramolecular Chiral Hydrogel on Cellular Behavior of Endothelial Cells Under High-Glucose-Induced Injury

In order to manage the impaired wound healing of diabetic chronic wounds induced by high-glucose-damaged endothelial cells, a glucose-induced injured endothelial cell model is established in this research to explore the regulatory effects of a supramolecular chiral hydrogel on injured endothelial cells. Cellular behaviors of endothelial cells under different culture conditions are evaluated by CCK-8, 5-ethynyl-2′-deoxyuridine (EdU) staining, adhesion, and Transwell assays. The expression levels of angiogenesis-related markers, including nitric oxide, endothelial nitric oxide synthase (eNOS), and vascular endothelial growth factor (VEGF), are also assessed by enzyme-linked immune sorbent assay (ELISA) and the polymerase chain reaction (PCR). The results demonstrate that the left-handed chirality of the supramolecular chiral hydrogel can promote cell proliferation and enhance the adhesion and migration ability of impaired endothelial cells. Moreover, enhanced nitric oxide synthesis and elevated expression of eNOS and VEGF are observed in cells in the left-handed chiral environment. Thus, the left-handed supramolecular chiral hydrogel can regulate the cellular behavior of high-glucose-injured endothelial cells, including cell proliferation, adhesion, migration, and angiogenesis, offering the potential to promote diabetic wound healing.

P2 purinergic receptor signaling and interleukin-1 synergistically induce interleukin-6 production in a human oral squamous carcinoma cell line

ObjectivesThe aim of this study was to investigate the inflammatory roles of P2 purinergic receptor (P2R) signaling in oral squamous cell carcinoma (OSCC). MethodsHuman OSCC cell lines HSC-2, Ca9-22, and HO-1-u-1 were stimulated with P2R agonists. The concentration of interleukin (IL)-6 in culture supernatants was measured using an enzyme-linked immune sorbent assay. Expression levels of messenger RNAs (mRNAs) were analyzed using reverse transcription polymerase chain reaction. Phosphorylation of intracellular signaling molecules was analyzed using western blotting. ResultsHSC-2 cells expressed the mRNAs for P2X4-6 and all P2YRs. ATP or ADP induced significantly greater production of IL-6 by HSC-2 cells. Ca9-22 cells expressed mRNAs for P2X4-6 and all P2YRs except P2Y4. ATP or ADP induced the production of IL-6 by Ca9-22 cells, but the IL-6 concentration was much lower than that in HSC-2 cells. Although HO-1-u-1 cells expressed the mRNAs for P2X4-6 and all P2YRs, ATP or ADP did not induce IL-6 production. The production of IL-6 by HSC-2 cells stimulated with adenine nucleotides was significantly inhibited by P2R antagonists and a p38 mitogen-activated protein kinase inhibitor, but not by extracellular signal-related kinase or c-Jun N-terminal kinase inhibitors. The proinflammatory cytokine IL-1 significantly augmented P2R-induced IL-6 production by HSC-2 cells via the nuclear factor-κB signaling pathway. ConclusionsThe present study suggests that P2Rs signaling and IL-1 synergistically induce chronic inflammation in OSCC. Because chronic inflammation is a well-known driving force of tumor progression, these results support therapeutic strategies that target P2Rs signaling in OSCC.

A case-control study to determination FBXW7 and Fetuin-A levels in patients with type 2 diabetes in Iraq.

diabetes mellitus (type 2) is a chronic disease characterized by hyperglycemia resulting either from a deficiency of insulin, or decrease insulin action or both. This study focused on determining the new marker F- box and WD repeat domain containing protein 7 (FBXW7) as well as Fetuin -A and their correlation with other parameters in Diabetes mellitus type 2. Sixty diabetes type2 patients were recruited from (National diabetes Center for treatment and Research) at Al Mustansriya University in Iraq during the period (from November 2018-March 2019) with an age range (37-69) years. The patient group was subdivided into two groups: group I (30) newly diagnosed T2DM and group II (30) T2DM (on metformin monotherapy), for comparison, 30 healthy individuals were included as a control. The protein levels (FBXW7, and Fetuin-A) were estimated using quantitative enzyme-linked immunesorbent assay (ELISA). Biochemical parameters; fasting serum glucose (FSG), and lipid profile were measured through spectrophotometrically. Insulin was determined using ELISA kit finally, HbA1c was determined using HPLC. Serum levels of FBXW7 protein were significantly higher (p-value = 0.01), whereas, there was no significant difference (p ˃ 0.05) in the level of Fetuin-A in group I as compared to control group. There was no significant difference (p ˃ 0.05) in the levels of both proteins who group II was compared with the control group. On the other hand, FBXW7 level were significantly lower in group II as compared to group I. Fasting blood sugar FSG, glycated hemoglobin HbA1c, body mass index BMI, waist- to-height ratio WHtR, and lipid profile were significantly elevated, except for high density lipoproteins, HDL was decreased in Group I as compared to control group, whereas insulin appeared non-significant (P > 0.05). FBXW7 Protein was negatively correlated to Fetuin -A, HOMA-IR, Insulin concentration, and BMI, whereas positive correlate to FSG, HbA1c, lipid profile, and atherogenic index. According to ROC analysis, the optimal cutoff value for FBXW7 protein was (1.455) ng/ml. Levels of FBXW7 and Fetuin-A proteins in sera of Diabetic patients were elevated. FBXW7 was significantly correlated with newly diagnosed patients. Metformin reduces FBXW7 and Fetuin-A levels.

Interleukin-1 family and serotonin transporter in first-episode, drug-naive major depressive disorder: A pilot study

Abnormalities of neuroinflammatory process and serotonergic system have been reported in major depressive disorder (MDD). However, most previous studies were performed in recurrent MDD and only a few studies explored the interaction of the two systems. This study examined both systems concurrently and their clinical relevance in first-episode drug-naive MDD. Thirty-four MDD patients and 34 age and gender matched healthy controls (HC) were recruited. Plasma concentrations of the cytokines of interleukin-1 (IL-1) family, including IL-1α, IL-1β, IL-1 receptor antagonist (IL-1Ra) and IL-1 receptor type 2 (IL-1R2) were measured using enzyme-linked immune-sorbent assays. The serotonin transporter (SERT) availability in midbrain, thalamus, caudate, and putamen was examined by single-photon emission computed tomography with 123I-ADAM. There were significantly lower concentrations of pro-inflammatory IL-1β and its inhibitor, IL-1R2 in MDD than HC. The SERT availability was at the same level between groups. A negative association between IL-1Ra concentration and the SERT availability in midbrain was observed in MDD but not in HC. Both IL-1β concentration and the SERT availability in caudate negatively correlated with depression severity and the effect of IL-1β was not moderated or mediated by the SERT. In conclusion, this study demonstrated the involvement of IL-1 family in the early stage of MDD, especially for IL-1β. SERT was not the main central target of altered IL-1β and these two systems might contribute to MDD by different mechanisms. The pathophysiology might be varied between early and recurrent MDD and tuning treatment strategies at different clinical stages might be needed.

Bio-nanocomposite based highly sensitive and label-free electrochemical immunosensor for endometriosis diagnostics application

In this research, for the first time, a bio-nanocomposites based highly sensitive and label-free electrochemical immunosensor is reported with the aim of endometriosis diagnostics application. Multiwalled carbon nanotube and magnetite nanoparticle (MWCNT-Fe3O4) was dispersed in chitosan (CS) to fabricate a bio-nanocomposite to immobilize very monoclonal specific antibody (via cross-linking using glutaraldehyde) for selective electrochemical immuno-sensing of carbohydrate antigen 19–9 (CA19-9), a potential biomarker for endometriosis diagnostics. Well-characterized Anti-AbsCA19-9/CS-MWCNT-Fe3O4 immune-electrode fabricated on glassy carbon electrode (GCE) successfully detect CA 19–9 and exhibited a high sensitivity as (2.55 µA pg−1 cm−1), a detection limit of 0.163 pg mL−1, detection range from 1.0 pg mL−1 to 100 ng mL−1. Our fabricated electrochemical AbsCA19-9/CS-MWCNT-Fe3O4 immunosensor performed CA19-9 sensing in physiological range and at a very level which suggest it application for early-stage diagnostics, diseases monitoring, and optimization of therapy. To claim the clinical application, our sensor was tested using real samples and sensing performance was validated using enzyme-linked immune-sorbent assay (ELISA). The results of the studies projected AbsCA19-9/CS-MWCNT-Fe3O4 electrochemical CA19-9 immunosensor as a potential and affordable alternate of conventional techniques like ELISA. We believe that our fabricated sensor can be the plane of a disease’s management program due to affordable, rapid, label-free, and sensitive detection of a targeted biomarker.

Hypoxic hUCMSC-derived extracellular vesicles attenuate allergic airway inflammation and airway remodeling in chronic asthma mice

BackgroundAs one of the main functional forms of mesenchymal stem cells (MSCs), MSC-derived extracellular vesicles (MSC-EVs) have shown an alternative therapeutic option in experimental models of allergic asthma. Oxygen concentration plays an important role in the self-renewal, proliferation, and EV release of MSCs and a recent study found that the anti-asthma effect of MSCs was enhanced by culture in hypoxic conditions. However, the potential of hypoxic MSC-derived EVs (Hypo-EVs) in asthma is still unknown.MethodsBALB/c female mice were sensitized and challenged with ovalbumin (OVA), and each group received PBS, normoxic human umbilical cord MSC-EVs (Nor-EVs), or Hypo-EVs weekly. After treatment, the animals were euthanized, and their lungs and bronchoalveolar lavage fluid (BALF) were collected. With the use of hematoxylin and eosin (HE), periodic acid-Schiff (PAS) and Masson’s trichrome staining, enzyme-linked immune sorbent assay (ELISA), Western blot analysis, and real-time PCR, the inflammation and collagen fiber content of airways and lung parenchyma were investigated.ResultsHypoxic environment can promote human umbilical cord MSCs (hUCMSCs) to release more EVs. In OVA animals, the administration of Nor-EVs or Hypo-EVs significantly ameliorated the BALF total cells, eosinophils, and pro-inflammatory mediators (IL-4 and IL-13) in asthmatic mice. Moreover, Hypo-EVs were generally more potent than Nor-EVs in suppressing airway inflammation in asthmatic mice. Compared with Nor-EVs, Hypo-EVs further prevented mouse chronic allergic airway remodeling, concomitant with the decreased expression of pro-fibrogenic markers α-smooth muscle actin (α-SMA), collagen-1, and TGF-β1-p-smad2/3 signaling pathway. In vitro, Hypo-EVs decreased the expression of p-smad2/3, α-SMA, and collagen-1 in HLF-1 cells (human lung fibroblasts) stimulated by TGF-β1. In addition, we showed that miR-146a-5p was enriched in Hypo-EVs compared with that in Nor-EVs, and Hypo-EV administration unregulated the miR-146a-5p expression both in asthma mice lung tissues and in TGF-β1-treated HLF-1. More importantly, decreased miR-146a-5p expression in Hypo-EVs impaired Hypo-EV-mediated lung protection in OVA mice.ConclusionOur findings provided the first evidence that hypoxic hUCMSC-derived EVs attenuated allergic airway inflammation and airway remodeling in chronic asthma mice, potentially creating new avenues for the treatment of asthma.

Effects of Duration of Diabetes and Diabetes Therapy (Metformin) on Fbxw7 Levels in Iraqi Type II Diabetic Patients

Background: This study concentrated on evaluation the FBXW7 levels and other markers in T2DM patients related to the duration of metformin monotherapy and duration of T2DM. Method: The current study includes 60 diabetic patients type II and30 control subjects. The groups of patients were subdivided in to two groups: (group I) (30) Newly diagnosed T2DM which also subdivided to two groups(&gt; 1 year, and &lt;1 year). Group II(30) T2DM (who were treated with metformin monotherapy) which also subdivided into two group (&gt; 1 year and &lt;1 year). In the present study FBXW7, FetuinA, pentraxin3, Nitric oxide and insulin were determined bythe enzyme linked immune sorbent assay (ELISA). Biochemical markers, fasting serum glucose, and lipide profile were determined through spectrophotometere technique in patients and control groups. Result: The levelsof glucose, HbA1c and IR significantly affected byincrease the duration of diseaseWhile each of fbxw7, pentraxin 3,NO, and fetuin A not influenced by increase duration of disease. There was nonsignificant effect of metformin therapy duration on levels of Fbxw7,fetuinA, pentraxin3 and lipid profile. While IR showed significant decrease when the duration of treatment increase. Conclusion: Fbxw7, fetuinA, pentraxin3 and NO arenot affected by long duration of therapy or by duration of disease while glucose, HbA1c, and IR are affected by increase duration of diseaseonly. Whereas insulin and IR are affected by both; duration of disease and therapy.

Exploration of bioactive constituents and immunoregulatory mechanisms of a hanshi-yufei formulation for treating COVID-19

Objective: The objective of this study was to characterize the chemical compounds of a Hanshi-Yufei formulation (HSYF; a modified formulation of a traditional Chinese medicine used for treating COVID-19) to elucidate the mechanism of action and to evaluate potential anti-inflammatory effects of HSYF. Materials and Methods: The chemical constituents of HSYF extract were characterized using UPLC-Q-TOF/MS. Subsequently, a set of TCM network pharmacology methods was applied to identify disease-associated genes and to predict target profiles and pharmacological actions associated with the constituents of HSYF. Then, the antiviral effects of HSYF on H1N1 were assessed in RAW264.7 cells using MTT assays. Expression levels of pro-inflammatory cytokines interleukin (IL)-6 and tumor necrosis factor (TNF)-α following infection of RAW264.7 cells with H1N1 were measured using an enzyme-linked immune sorbent assay (ELISA), and expression levels of inflammatory-related factors were detected using western blotting. Results: In total, 165 chemical constituents (including glycosides, tannins, volatile oils, amino acids, triterpenoids, polyphenols, phenylpropanoids, sesquiterpenes, alkaloids, and flavonoids, among others) were tentatively identified in HSYF. Network pharmacology demonstrated that HSYF can regulate immunomodulatory- and anti-inflammatory-related targets of multiple pathways through its active ingredients, suggesting potential anti-COVID-19 effects. Furthermore, cell viability assays and ELISA showed that HSYF significantly inhibited H1N1 replication in RAW64.7 cells and markedly reduced expression of pro-inflammatory cytokines TNF-α and IL-6 at the proteins level. Conclusions: The results of the present study help improve our understanding of the therapeutic effects of HSYF in COVID-19 treatment from multi-level perspectives.

Chrysoeriol promotes functional neurological recovery in a rat model of cerebral ischemia

Background: Stroke is among the leading causes of death worldwide, but current treatment options for cerebral ischemia are limited. Chrysoeriol is a natural isolated agent with bioactive pharmacological properties. Materials and Methods: Sprague–Dawley rats were subjected to transient middle cerebral artery occlusion (MCAO) surgery and chrysoeriol was administrated for the subsequent 3 days. Neurological deficit scores, apoptosis, oxidant stress, and inflammatory cytokines were then measured and the effects on the Wnt/β-catenin pathway were evaluated. Results: Behavioral tests revealed that chrysoeriol promoted recovery from neurological deficits in rats. Hematoxylin and eosin staining demonstrated that chrysoeriol alleviated neurological damage. Triphenyl tetrazolium chloride staining demonstrated that chrysoeriol reduced the area of ischemia. An enzyme-linked immune sorbent assay (ELISA) demonstrated that chrysoeriol inhibited excessive pro-inflammatory cytokine production (tumor necrosis factor-α, interleukin [IL]-1β, and IL-6) and regulated oxidative stress (malondialdehyde, superoxide dismutase, and glutathione). Caspase-3 immunofluorescence and a terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay showed that chrysoeriol reduced neuronal apoptosis. The ELISA assay and Nissl staining showed that chrysoeriol promoted neuronal survival and growth. Western blotting and an immunofluorescence assay demonstrated that the Wnt/β-catenin signaling pathway plays a crucial role in the neuronal effects of chrysoeriol. The Wnt inhibitor, Dickkopf-related protein-1, validated the role of the Wnt/β-catenin signaling pathway in the neuroprotective effects of chrysoeriol. Conclusion: Chrysoeriol exerts neuroprotective effects in a rat model of MCAO. These effects are mediated by the Wnt/β-catenin signaling pathway.

A "sandwich" cell culture platform with NIR-responsive dynamic stiffness to modulate macrophage phenotypes.

Considering the key roles of macrophages in tissue repair and immune therapy, designing smart biomaterials able to harness macrophage phenotypes on demand during the healing process has become a promising strategy. Here, a novel "sandwich" cell culture platform with near-infrared (NIR) responsive dynamic stiffness was fabricated to polarize bone marrow-derived macrophages (BMDMs) in situ for revealing the relationship between the macrophage phenotype and substrate stiffness dynamically. Under NIR irradiation, calcium ions (Ca2+) diffused through the middle layer of the IR780-mixed phase change material (PCM) due to the photothermal effect of IR780, resulting in an increase of hydrogel stiffness in situ by the crosslinking of the upper layer of the hyaluronic acid-sodium alginate hydrogel (MA-HA&SA). The up-regulation of inducible nitric oxide synthase (iNOS) and tumor necrosis factor-α (TNF-α) was quantified by immunostaining and enzyme-linked immune sorbent assay (ELISA), respectively, indicating the transformation of macrophages from the anti-inflammatory to pro-inflammatory phenotype under dynamic stiffness. The nuclear Yes-associated-protein (YAP) ratio positively correlated with the shift of the macrophage phenotype. The modulation of macrophage phenotypes by stiffness-rise without the stimuli of cytokines offers an effective and noninvasive strategy to manipulate immune reactions to achieve optimized healing or therapeutic outcomes.

Evaluation of The Relationship among The Levels of SIRT1 and SIRT3 with Oxidative Stress and DNA Fragmentation in Asthenoteratozoospermic Men.

BackgroundReactive oxygen species (ROS) play a crucial role in etiology of DNA fragmentation and lipid peroxi- dation in sperm, leading to infertility in men. The silent information regulators SIRT1 and SIRT3 are members of the sirtuins protein family known to be involved in cancer genetics, aging and oxidative stress responses. The aim ofthis study is to determine the correlation between SIRT1 and SIRT3 with antioxidants, oxidative stress biomarkers, and DNA fragmentation in the semen of asthenoteratozoospermic and normozoospermic men.Materials and MethodsIn this case-control study, after spermogram analysis the specimens were divided into two groups, normozospermic (n=40) and asthenoteratozoospermic (n=40), according to World Health Organization (WHO) standards. Sperm DNA fragmentation was evaluatedusing the sperm chromatin dispersion (SCD) test.Catalase activity was measured using the Aebi spectrophotometeric method. Total antioxidant capacity (TAC) level and super- oxide dismutase (SOD) activitywere measured by using commercially available colorimetric assays. Enzyme-linked immune sorbent assay (ELISA) was used to measure SIRT1 and SIRT3 protein levels of seminal plasma. Malondial- dehyde (MDA) level in seminal plasma was determined by high-performance liquid chromatography (HPLC).ResultsThe asthenoteratozoospermic group had significantly lower catalase and SOD activities and TAC levels in comparison with the normozoospermic group (P<0.001).The percentage of DNA fragmentation and MDA level in the asthenoteratozoospermic group were remarkably higher than in the normozoospermic group. The SIRT1 and SIRT3 protein levels in seminal plasmawere remarkably lower in asthenoteratozoospermic group than the normozoospermic group (P<0.001).ConclusionThe results of this study suggest that SIRT1 and SIRT3 protein levels are negatively correlated with oxidative stress and DNA fragmentation in semen. The low levels of SIRT1 and SIRT3 in asthenoteratozoospermic men may lead to an increase in oxidative stress, DNA fragmentation, and lipid peroxidation that eventually result in immotile and immature spermatozoa (asthenoteratozoospermia).

Safety and immunogenicity of SURE RAB™, an inactivated cell culture derived rabies vaccine: A comparative, phase III clinical trial

IntroductionRabies is a viral zoonotic disease widespread across the world. In India, various cell culture vaccines are available for pre and post exposure prophylaxis (PEP) but are not sufficient to meet the rising demand. The present study evaluated the safety and immunogenicity of Rabies vaccine Human I.P. (Brand name: SURE RAB™) in PEP and demonstrated its non-inferiority to already approved rabies vaccine (Brand name: VERORAB). Material and methodsIt was a phase-III randomized, open label, comparative, single centre clinical trial in post exposure subjects. Safety and immunogenicity were evaluated at Day 0, 14 and 45 ± 7 post vaccination. Day 14 serum samples were analyzed by Enzyme Linked Immunesorbent assay (IgG ELISA, Bio-Rad) and Day 0, 14 and 45 ± 7 serum were tested by Rapid Fluorescent Focus Inhibition Test (RFFIT). Paired t-test was applied to compare the results of Rabies virus neutralizing antibody (RVNA). The severity of adverse reactions was measured on a scale of excellent, good, fair and bad; p-value (p < 0.05) was considered as statistically significant. ResultsAll the subjects achieved a protective titer value between 0.5 and 9.0 IU/ml by Day 14 tested by ELISA and significant rise in the antibody titer in all the groups when tested after 45 days. Statistically significant p-value (p < 0.001) observed with RFFIT test indicated biological potency of rabies vaccine. Adverse events and safety was comparable statistically between three groups (p ​= ​0.886) and Group I ​+ ​II combined versus Group III (p = 0.495). ConclusionThe study results conclusively demonstrate that SURE RAB™ is comparable to VERORAB in terms of safety and immunogenicity and can be used for PEP in rabies.

Study of Golgi protein 73 in patients with hepatitis C virus-related hepatocellular carcinoma

ObjectiveThe aim of the present work is to investigate the expression of Golgi protein 73 (GP73) in serum in patients with hepatocellular carcinoma (HCC) and determine its efficacy as a screening test in early detection of HCC.BackgroundSerum GP73 is a promising biomarker for detection of HCC.Patients and methodsThis case–control study involved 30 patients with HCC, 30 patients with liver cirrhosis, and 15 healthy controls. Clinical examination, abdominal ultrasonography, and triphasic computed tomography for focal lesion were performed. Liver function tests were performed using clinical autoanalyzer, serum α-fetoprotein (AFP) was measured using enzyme-linked immune-sorbent assay method, and GP73 was determined by enzyme-linked immune-sorbent assay kit for GP73. Data were collected and statistically analyzed.ResultsGP73 was highly significantly higher in HCC group than cirrhotic and control groups. The sensitivity of GP73 in diagnosis of HCC was 83% and the specificity was 74%, at a cutoff point of 111 ng/ml, elicited from the receiver operator characteristic curve, with very good area under curve of 0.909, whereas that the sensitivity of AFP was 36.7% and the specificity was 100% at a cutoff point of 400 ng/ml, elicited from the receiver operator characteristic curve, with very good area under curve of 0.903.ConclusionGP73 is highly associated with HCC and is more sensitive than AFP for early detection of HCC.

Packaging fortified with Natamycin nanoparticles for hindering the growth of toxigenic Aspergillus flavus and aflatoxin production in Romy cheese.

Objective:This study assessed the effect of cellulose sheets fortified with Natamycin-loaded alginate nanoparticles on the growth of toxigenic Aspergillus flavus and aflatoxin production on the superficial layer of Egyptian Romy cheese after 12 weeks of maturation.Materials and Methods:Toxigenic A. flavus (GenBank accession No. MT645073) was inoculated into the outer surface of Egyptian Romy cheese (at 5 log CFU/gm) and wrapped with a cellulose sheet fortified with Natamycin-loaded alginate nanoparticles. Unwrapped control contaminated Romy wheels were made as well as non-contaminated wrapped cheese wheels for sensory evaluation. Romy cheese wheels were stored at a temperature similar to commercial methods for 12 weeks. Fungal counts were enumerated during this time, and enzyme-linked immune sorbent assay detected aflatoxin after the 4th week of maturation storage.Results:In cheese samples covered with cellulose sheets containing Natamycin-loaded alginate nanoparticles, the fungal count was reduced by 2 log approximately in contrast to control samples after the 2nd week of storage. However, within the 8th week of storage, the greatest significant reduction (p < 0.05) was seen where fungal growth was hindered entirely to the end of the ripening period. The mean values for taste, color, flavor, and overall acceptability were 4, 4.7, 4.09, and 4.3, respectively. Furthermore, in the treated samples, the total aflatoxin concentration was decreased by 78.6% relative to the untreated control one.Conclusion:Using cellulose sheets fortified with Natamycin-loaded alginate nanoparticles in Egyptian Romy cheese wrapping could be an effective way of controlling A. flavus and subsequent aflatoxin production without influencing the typical taste, color, flavor, and overall appearance of traditional Romy cheese.

MiR-22 Host Gene Enhances Nuclear Factor-kappa B Activation to Aggravate Hypoxia-induced Injury in AC16 Cardiomyocytes

Myocardial infarction (MI) is a severe life-threatening disease caused by acute and persistent ischemia and hypoxia and eventually leads to heart failure and sudden death. Long noncoding RNAs (lncRNAs) play significant roles in the pathology, diagnosis, and development of various cardiovascular diseases, including MI. This study aimed to explore the effect and molecular mechanism of lncRNA miR-22 host gene (MIR22HG) on hypoxia-induced injury in AC16 cardiomyocytes. The expression of MIR22HG and miR-24 in hypoxia-treated AC16 cardiomyocytes was detected by quantitative real-time polymerase chain reaction. Cell viability, lactate dehydrogenase release, levels of aspartate aminotransferase (AST) and creatine kinase-MB (CK-MB), and apoptosis were detected by Cell Counting Kit-8, lactate dehydrogenase (LDH) release assay, commercial enzyme-linked immune sorbent assay kits, and flow cytometry analysis, respectively. The protein levels of nuclear factor-kappa B (NF-κB) p65 and cytoplasmic inhibitor of kappa B alpha (IκBα) and phosphorylated IκBα were detected by western blot. Results showed that hypoxia treatment decreased viability and increased MIR22HG expression in AC16 cardiomyocytes. MIR22HG overexpression aggravated hypoxia-induced viability reduction, leakage of myocardial injury markers LDH, AST, and CK-MB, and apoptosis in AC16 cardiomyocytes, while MIR22HG knockdown elicited the reverse effects. MIR22HG overexpression enhanced NF-κB activation in hypoxia-treated AC16 cardiomyocytes. Inhibition of NF-κB pathway impaired the effects of MIR22HG overexpression on hypoxia-induced injury in AC16 cardiomyocytes. Moreover, MIR22HG knockdown inhibited the NF-κB pathway by upregulating miR-24 in AC16 cardiomyocytes. Inhibition of miR-24 resisted the effects of MIR22HG silencing on hypoxia-induced injury in AC16 cardiomyocytes. In conclusion, MIR22HG overexpression aggravated hypoxia-induced injury in AC16 cardiomyocytes via enhancing NF-κB activation by targeting miR-24.

Effects of intraperitoneal and intracerebroventricular injection of cinnamaldehyde and yohimbine on blood glucose and serum insulin concentrations in ketamine-xylazine induced acute hyperglycemia.

This study was designed to investigate the effects of peripheral [intraperitoneal (IP)] and central [intracerebroventricular (ICV)] administration of cinnamaldehyde on concentrations of blood glucose and serum insulin in the acute hyperglycemia induced by ketamine/xylazine. Yohimbine (a α2-adrenoceptor antagonist) was used alone and in combination with cinnamaldehyde to explore the α2-adrenergic receptor contribution. A total of 48 rats were divided into eight groups with six rats in each for IP administration of normal saline, vehicle, cinnamaldehyde (25.00, 50.00 and 100 mg kg-1), yohimbine (0.50 and 2.00 mg kg-1) and cinnamaldehyde plus yohimbine. These rats were used again for ICV administration 15 days after the completion of IP experiment. During this 15 days period, the lateral ventricle of the brain was surgically cannulated for ICV administration of normal saline, vehicle, cinna-maldehyde (25.00, 50.00 and 100 µg per rat), yohimbine (5.00 and 20.00 µg per rat) and cinnamaldehyde plus yohimbine. Blood glucose levels were measured from tail blood using a glucometer and serum insulin concentrations were determined via enzyme-linked immune-sorbent assay kit. The increased levels of blood glucose and the decreased concentrations of serum insulin were significantly decreased and increased, respectively, by separate and combined IP and ICV administrations of cinnamaldehyde and yohimbine. The systemic effects of these chemical compounds were significantly greater than the central ones. Based on the results, it can be argued that cinnamaldehyde has a potential to induce anti-hyperglycemic and antihypoinsulinemic effects. Peripheral and central α2-adrenegic receptors might be involved in these effects of cinnamaldehyde.

Artificial intelligence for COVID-19 and future pandemics: A mini-review

The pandemic of severe acute respiratory syndrome coronavirus 2 has spread very quickly all over the world and has become an unparalleled public health crisis. This unforeseen and exceptional situation has instigated a wave of research to investigate the virus, track its spread, and study the disease it causes. Current methods of diagnosis and monitoring largely rely on polymerase chain reactions and enzyme-linked immunesorbent assay methods. In this hour of crisis, researchers are looking for new technologies to monitor and control such disease outbreaks. Artificial intelligence (AI) is one such technology. Being an evidence-based tool, this technology has the potential to upgrade our disease management strategies and help us to restrict the spread of such diseases. AI can play an effective role in tracking the spread of diseases, screening of the population, identifying patients and developing treatments of diseases. Through this review, we aim to analyze the role of AI in the diagnosis, monitoring and treatment of diseases like coronavirus disease 2019, with most recent updates and assess the prospects of this technology in the management of such diseases.