Functional characterization and in vitro screening of Fructobacillus fructosus MCC 3996 isolated from Butea monosperma flower for probiotic potential.

The fructophilic bacterium Fructobacillus fructosus MCC 3996 described in the present investigation was isolated from the nectar of Butea monosperma flower and evaluated in vitro for the manifestation of probiotic features. The strain utilizes fructose faster than glucose and is capable to grow in the range of 1-35% fructose concentration (optimum 5% w/v) and thus denotes its fructophilic nature. In vitro assessments of the strain have examined for the endurance in acidic environment/gastric juice, the better auto-aggregation ability even in the presence of hydrolytic enzymes, co-aggregation with pathogenic bacteria, hydrophobicity properties and no haemolytic activity to elucidate its feasible probiotic use. The significant antagonistic activity against several detrimental bacteria, despite lacking the bacteriocin secretion, is an astonishing feature. Owing to the indigenous origin of the isolate, it could be used as a probiotic, starter culture, and/or the active ingredient of food formulation may contribute to improve the desirable fermentation, long-term storage and nutritional benefits of foods especially rich in fructose. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provided in vitro evidence that Fructobacillus fructosus MCC 3996 have endurance in acidic gastric juice, better co-aggregation, auto-aggregation properties, splendid antagonistic activities against several bacteria involved in food spoilage/human infections, pertinent antibiotic susceptibility profile and no haemolytic activity. Also, F. fructosus have the capability to survive in the appreciable amount of fructose, and this advocates that the strain could be used as starter culture and/or the active ingredient of fructose-rich foods. The current in vitro study provided a strong basis for further in vivo research to identify the health beneficial characteristics of F. fructosus and its potential could be effectively utilized as health-boosting ingredient in food and pharmaceutical industries.

Installing oncofertility programs for common cancers in limited resource settings (Repro-Can-OPEN Study): An extrapolation during the global crisis of Coronavirus (COVID-19) pandemic.

PURPOSE: The state of limited resource settings that Coronavirus (COVID-19) pandemic has created globally should be taken seriously into account especially in healthcare sector. In oncofertility, patients should receive their fertility preservation treatments urgently even in limited resource settings before initiation of anticancer therapy. Therefore, it is very crucial to learn more about oncofertility practice in limited resource settings such as in developing countries that suffer often from shortage of healthcare services provided to young patients with cancer. METHODS: As an extrapolation during the global crisis of COVID-19 pandemic, we surveyed oncofertility centers from 14 developing countries (Egypt, Tunisia, Brazil, Peru, Panama, Mexico, Colombia, Guatemala, Argentina, Chile, Nigeria, South Africa, Saudi Arabia, and India). Survey questionnaire included questions on the availability and degree of utilization of fertility preservation options in case of childhood cancer, breast cancer, and blood cancer. RESULTS: All surveyed centers responded to all questions. Responses and their calculated oncofertility scores showed different domestic standards for oncofertility practice in case of childhood cancer, breast cancer, and blood cancer in the developing countries under limited resource settings. CONCLUSIONS: Medical practice in limited resource settings has become a critical topic especially after the global crisis of COVID-19 pandemic. Understanding the resources necessary to provide oncofertility treatments is important until the current COVID-19 pandemic resolves. Lessons learned will be valuable to future potential worldwide disruptions due to infectious diseases or other global crises.

Effect of metformin on thyroid function tests in patients with subclinical hypothyroidism: an open-label randomised controlled trial.

PURPOSE: Though most of the observational studies have shown that metformin can reduce serum thyroid stimulating hormone (TSH) level in patients of hypothyroidism with diabetes or polycystic ovarian disease, randomised controlled trials are sparse. The primary objective of this study was to evaluate the effect of metformin on thyroid function tests (TSH, free T4, and free T3) in patients with subclinical hypothyroidism (SCH). METHODOLOGY: In this open label, parallel arm, randomised controlled trial, 60 patients of SCH (TSH 5.5-10 mIU/L) were randomised to either metformin group (1500 mg/day) or control group. RESULT: A total of 46 patients (23 in each group) completed the study and no significant difference in serum TSH, free T4 or free T3 was found in between the 2 groups. Neither there was any significant change in serum TSH, free T4 or free T3 (pre and post 6 months) within the individual groups. However, the rate of normalisation of serum TSH in patients with negative thyroid antibody was significantly higher than patients with positive thyroid antibody (71.4% vs. 18.8%; P = 0.026) in metformin group in post hoc analysis. Fasting plasma glucose, serum high-density lipoprotein and indices of insulin sensitivity significantly improved in metformin group. Four patients (17%) had mild gastrointestinal adverse effects in the metformin group. CONCLUSION: We did not find any significant change in thyroid function test in patients with SCH with metformin therapy.

Human filarial proteins attenuate chronic colitis in an experimental mouse model.

Encouraged by our earlier results of promising therapeutic effect of filarial recombinant proteins BmALT2, BmCys and WbL2 individually in the mouse model of acute ulcerative colitis, in this study, these proteins have been explored individually and in different combinations for their therapeutic potential in dextran sulphate sodium (DSS)-induced chronic colitis mice. These mice, treated with filarial proteins, showed reduced disease parameters including body weight loss, disease activity index, macroscopic and histopathological scores of colon and myeloperoxidase activity in colonic mucosa. Among various treatment schemes, rBmALT2 + rBmCys which showed most pronounced therapeutic implication was found to downregulate the mRNA expressions of IFN-γ and TNF-α and upregulate IL-10 and TGF-ß expression in the splenocytes. Also, increase in level of IgG1 and IgG2a isotypes in the sera of rBmALT2 + rBmCys-treated colitis mice was noted. Activated NF-κB level was found to be reduced in the colon of treated colitis mice compared to untreated one. In conclusion, filarial proteins in combination have been shown to improve the clinicopathologic status of chronic colitis through suppression of pro-inflammatory immune response most possibly in NF-κB-dependent manner. We propose this therapeutic strategy to be tested further to be considered as an effective option in chronic colitis.

Evaluation of Ageratum conyzoides in field scale constructed wetlands (CWs) for domestic wastewater treatment.

Ageratum conyzoides were evaluated in field scale subsurface flow constructed wetlands (CWs) to quantify its nitrogen (N) and phosphorus (P) uptake and compare with wetland plants (Pistia stratiotes, Typha latifolia and Canna indica). The two-field scale subsurface flow CWs, located in the International Crops Research Institute for Semi-Arid Tropics, received wastewater from an urban colony. The CW1 and CW2 had the same dimensions (length:10 m, width:3 m, total depth:1.5 m and sand and gravel:1 m), similar flow rates (3 m3/d), hydraulic loading rates (HLRs-10 cm/d) and hydraulic retention time (HRT-5 days) from July 2014-August 2015. The vegetation in both CWs consisted of Pistia stratiotes, Typha latifolia, Canna indica, and Ageratum conyzoides, respectively. The CW1 (% reduction with respect to concentrations) reduced total suspended solids (TSS) (68%), NH4-N (26%), NO3-N (30%), soluble reactive P (SRP) (20%), chemical oxygen demand (COD) (45%) and fecal coliforms (71%), while the CW2 (%-reduction with respect to concentrations) reduced TSS (63%), NH4-N (32%), NO3-N (26%), SRP (35%), COD (39%) and fecal coliforms (70%). Ageratum conyzoides can be used in combination with Pistia stratiotes, Typha latifolia and Canna indica to enhance removal of excessive N, P and fecal coliforms from domestic wastewater.

Biological Activities and Identification of Bioactive Metabolite from Endophytic Aspergillus flavus L7 Isolated from Aegle marmelos.

Aegle marmelos, a well-known Indian plant with medicinal and religious importance, has been extensively used in Indian traditional medicine. The present study aimed to isolate, identify, and evaluate the biological activities of endophytic fungi from A. marmelos. One of the isolates, labeled as L7, was identified as Aspergillus flavus using morphology and ITS gene sequence. Total phenolic and flavonoid contents in the culture filtrate were found to be 65.77 mg GAE/ml and 158.33 mg quercetin/ml of crude extract, respectively. The extract showed excellent antimicrobial activity against common human bacterial and fungal pathogens. The test extract at 700 µg/ml, which notably reduced the concentration of DPPH-free radical as percent DPPH scavenging activity, was found to be the highest (64.53 %). The extract, at the concentration of 2 mg/ml, produced 70 % inhibition of hemolysis of RBCs compared to 78 % produced by standard drug (Ibuprofen). Chemical profiling of the fermented extract using TLC followed by UV and FTIR revealed the presence of flavonoids. The HPLC analysis confirmed the presence of bioflavonoid rutin in the extract. To the best of our knowledge, this is the first report on production of bioactive flavonoid by endophytic Aspergillus flavus obtained from A. marmelos and its pharmaceutical potential. In conclusion, the endophytic Aspergillus flavus obtained from the A. marmelos could be explored as an economic and potential natural resource with diverse pharmaceutical and biological activities.

Antimicrobial potential of metabolites extracted from bacterial symbionts associated with marine sponges in coastal area of Gulf of Mannar Biosphere, India.

UNLABELLED: Marine coastal areas of India have vast diversity of sponges which harbours many endosymbiotic bacteria which are the source of many potential antimicrobial metabolites. This study focuses the screening and characterization of drug-producing bacteria symbiotically which are associated with marine sponges collected from Gulf of Mannar, South Coast India. Six different sponges were collected and they were identified on the basis of their morphology. The drug-producing isolates were screened by agar overlay method towards various clinical strains. The secondary metabolites were characterized and were found to be quinones, alkaloids, flavanoids and flavonyl glycosides. The metabolites showed significant inhibitory properties against clinical strains that were further identified as chromophoric and fluorophoric in nature. Ethyl acetate extracts of chromophore and floureophore substances showed significant inhibitory properties against Methicillin resistant Staphylococcus aureus (MRSA) and Salmonella typhi respectively. 16S rRNA gene sequencing of theses isolates revealed that chomophore-producing strain were closely related to Pseudomonas spp. RHLB12, isolated from Callyspongia spp. and floureophore-producing bacteria was related to Bacillus licheniformis T6-1 which was isolated from Haliclona spp. Hence, our study demonstrated that antimicrobial metabolites extracted from symbiotic bacteria associated with marine sponges have high therapeutic potential against many bacterial pathogens including multidrug-resistant strains. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study demonstrating antimicrobial potential of flurophoric and chromophoric metabolites extracted from bacterial biosymbionts associated with marine sponges. Our study has significant scope as Indian coastal area especially harbours vast varieties of sponges with novel secondary metabolites-producing organisms. The natural metabolites extracted from sponge-derived bacteria pave novel therapeutic remedy against various pathogens when most of them are emerged as extreme drug resistant superbugs.

Arterial thrombosis in "protein C deficiency"--a rare event.

We report here a case of a 35 year old male with protein C deficiency who presented with acute right sided hemiparesis with right sided facial palsy due to cerebral arterial thrombosis. He was treated with anticoagulation therapy and improved. This case is interesting as arterial thrombosis is rarely observed event in protein C deficiency.

Evaluation of surgically treated primary spinal cord tumors in a single Indian institution: A case series study of 178 patients.

Introduction: Primary spinal cord tumors are rare tumors that are usually heterogeneous having variable histopathological subtypes. Rapidly, growing space-occupying lesions in the spinal canal cause severe loss of function. This study aimed to analyze all adult patients diagnosed with primary spinal cord tumors and to describe their location, symptoms, histopathological types, surgical outcome, complications, recurrence in a single institution, and prognostic factors. Materials and Methods: The study was a retrospective study of 178 patients with spinal tumor who underwent surgical intervention at the department of neurosurgery at a tertiary care institute from 2014 to 2021. Patient demographic characteristics, symptoms, radiological features, spinal level of tumor, spinal compartment, preoperative and postoperative McCormick grade, and complications were recorded, and a comparison of outcomes after surgical intervention was done. Results: In our study, 97 patients were males and the rest of the 81 patients were females. The mean age at the time of surgery was 43.5 years. The thoracic region of the spinal canal was the most involved with 67 (37.64%) cases, whereas intradural extramedullary was the most common (65.73%). Schwannoma was the most common histopathological subtype (46.06%). Total excision was achieved in 154 cases. A total of 37 patients were in Grades 4 and 5 of modified McCormick's grade preoperatively. However, postoperatively, there were only 17 patients in this group. Two patients with cervical intramedullary tumor expired in the postoperative period. Conclusion: Early surgical intervention with sound microneurosurgical skills gives good outcomes independent of the type and location of tumor. Better outcomes are seen in extradural tumors, early surgical intervention, tumors better amenable to total resection, and in patients with better preoperative neurological status.

Hearing loss in British Army musicians.

BACKGROUND: Military musicians increasingly fulfil front line roles. Existing literature suggests that musicians are at greater risk of hearing loss than the general population. It is important to ascertain whether these findings apply to military musicians if they are to be placed in operational acoustically hazardous environments. AIMS: To compare hearing loss between groups of British Army musicians and with their administrative (non-musician) counterparts. METHODS: Routine health surveillance audiometric data for a group of military musicians were compared with military administrative staff during the first 8-12 years of their careers. Structured interviews were used to identify those meeting the study inclusion criteria (8 year minimum service, no significant, relevant medical history, no previous extraordinary loud noise exposure, service within study employment group throughout military career), and to clarify the instrument played. RESULTS: Eighty-four musicians and 44 administrators participated. Using the Health & Safety Executive's (HSE's) age- and gender-correcting system, 12 (14%) musicians and 9 (20%) non-musicians had a deterioration in hearing. The overall odds ratio for developing hearing loss in musicians was 0.65 (95% CI 0.25-1.68). Compared with woodwind instrumentalists, the odds ratio for hearing loss in brass instrumentalists was 2.00 (95% CI 0.85-3.44) and for percussionists 1.83 (95% CI 0.77-2.90). CONCLUSIONS: The findings suggest that military musicians are at no greater risk of hearing loss than their administrative counterparts after 8-12 years in service. This may represent a healthy-worker effect. There is no statistically significant differential risk of hearing loss between different instrumental groups.

Multicenter matched-pair study comparing BACT/ALERT® MP reagent systems for the recovery of mycobacteria from specimens other than blood.

The BACT/ALERT® MP Reagent System is a broth culture medium for optimal detection and recovery of mycobacteria from clinical samples. The MP formulation was recently modified to improve detection and recovery times. A multicenter prospective matched pair study design was conducted to validate the performance of improved MP (MP-I) versus current MP (MP-C) bottles utilizing nonsterile and normally sterile samples, except blood, from patients suspected of having mycobacterial infections. A total of 1488 clinical samples were collected to obtain 212 mycobacteria samples by either or both MP culture bottles. MP-I and MP-C sensitivities were 86.6% and 81.4%, respectively, but the difference was not significant (P = 0.163) while specificities were 96.8% and 93.8%, respectively, and that difference was significant (P = 0.002). Overall recovery was 94.34% for MP-I and 88.68% for MP-C (recovery was 100% for both bottles with 52 seeded samples). Overall performance of MP-I was better than MP-C for sensitivity, specificity, and recovery.

Prediction score for prolonged hospital stay in meconium aspiration syndrome: A multicentric collaborative cohort of south India.

BACKGROUND AND OBJECTIVE: With improved survival in neonates with meconium aspiration syndrome (MAS), the focus is currently on mitigating the morbidities. The objective of this study was to predict factors determining prolonged hospital stay in neonates with MAS. MATERIALS AND METHODS: It was a retrospective cohort from five centers of south India between 2018 and 2020. Neonates ≥35 weeks of gestation admitted to neonatal intensive care unit with the diagnosis of MAS and requiring oxygen beyond 24 h of life were included in the study. The morbidities in the neonates with stay ≤7 days (short stay) were compared with >7 days (prolonged stay). Logistic regression by the backward stepwise method was used for predictive score creation. RESULTS: Out of 347 neonates with MAS discharged home, 103 (29%) had a short stay and 244 (71%) had prolonged stay. The primary support beyond O2 (continuous positive airway pressure/mechanical ventilation) (42% vs. 83%, p < 0.001), fractional inspired oxygen (FiO2 ) at 1 h >30% (45% vs. 87%, p < 0.001), hypoxic ischemic encephalopathy (HIE) stage 2 or 3 (1% vs. 27%, p < 0.001), moderate-severe persistent pulmonary artery hypertension (PPHN) (3% vs. 31%, p < 0.001) were independent factors associated with prolonged stay on logistic regression. A prediction model was devised using weighted scores of these four associated morbidities. The clinical score thus developed had 83% sensitivity, 68% specificity for the prediction of prolonged stay (area under curve: 82%, 95% confidence interval [78-87], p < 0.001). CONCLUSION: More than two-thirds of neonates with MAS had prolonged stay. The primary support beyond oxygen, FiO2 requirement >30%, Moderate to severe PPHN, HIE stage 2 or 3 were predictive of prolonged stay in neonates with MAS.

The small-nucleolar RNAs commonly used for microRNA normalisation correlate with tumour pathology and prognosis.

BACKGROUND: To investigate small-nucleolar RNAs (snoRNAs) as reference genes when measuring miRNA expression in tumour samples, given emerging evidence for their role in cancer. METHODS: Four snoRNAs, commonly used for normalisation, RNU44, RNU48, RNU43 and RNU6B, and miRNA known to be associated with pathological factors, were measured by real-time polymerase chain reaction in two patient series: 219 breast cancer and 46 head and neck squamous cell carcinoma (HNSCC). SnoRNA and miRNA were then correlated with clinicopathological features and prognosis. RESULTS: Small-nucleolar RNA expression was as variable as miRNA expression (miR-21, miR-210, miR-10b). Normalising miRNA PCR expression data to these recommended snoRNAs introduced bias in associations between miRNA and pathology or outcome. Low snoRNA expression correlated with markers of aggressive pathology. Low levels of RNU44 were associated with a poor prognosis. RNU44 is an intronic gene in a cluster of highly conserved snoRNAs in the growth arrest specific 5 (GAS5) transcript, which is normally upregulated to arrest cell growth under stress. Low-tumour GAS5 expression was associated with a poor prognosis. RNU48 and RNU43 were also identified as intronic snoRNAs within genes that are dysregulated in cancer. CONCLUSION: Small-nucleolar RNAs are important in cancer prognosis, and their use as reference genes can introduce bias when determining miRNA expression.

Continuous low-dose cyclophosphamide and methotrexate combined with celecoxib for patients with advanced cancer.

BACKGROUND: Combined therapy of metronomic cyclophosphamide, methotrexate and high-dose celecoxib targeting angiogenesis was used in a phase II trial. METHODS: Patients with advanced cancer received oral cyclophosphamide 50 mg o.d., celecoxib 400 mg b.d. and methotrexate 2.5 mg b.d. for two consecutive days each week. Response was determined every 8 weeks; toxicity was evaluated according to CTC version 2.0. Plasma markers of inflammation, coagulation and angiogenesis were measured. RESULTS: Sixty-seven of 69 patients were evaluable for response. Twenty-three patients had stable disease (SD) after 8 weeks, but there were no objective responses to therapy. Median time to progression was 57 days. There was a low incidence of toxicities. Among plasma markers, levels of tissue factor were higher in the SD group of patients at baseline, and levels of both angiopoietin-1 and matrix metalloproteinase-9 increased in the progressive disease group only. There were no changes in other plasma markers. CONCLUSION: This metronomic approach has negligible activity in advanced cancer albeit with minimal toxicity. Analysis of plasma markers indicates minimal effects on endothelium in this trial. These data for this particular regimen do not support basic tenets of metronomic chemotherapy, such as the ability to overcome resistant tumours by targeting the endothelium.

Validated TLC method for simultaneous quantitation of kutkoside and picroside-I from Kutki extract.

INTRODUCTION: The two iridoid glycosides kutkoside and picroside-I are the active hepatoprotective principles of Picrorhiza kurroa Royle ex Benth (Scrophulariaceae), commonly known as Kutki. Quantitation of these phytoconstituents is important for the routine quality control of Kutki extract. OBJECTIVE: To develop and validate a simple, precise and rapid thin-layer chromatography (TLC) method for the simultaneous quantitation of kutkoside and picroside-I in Kutki extract. METHODOLOGY: The analysis was performed on a TLC precoated silica gel 60 F(254) plate with ethyl acetate:methanol:glacial acetic acid:formic acid (25:5:1:1, v/v/v/v) as mobile phase. Densitometric evaluation of kutkoside and picroside-I was carried out at 265 nm and the mobile phase showed good resolution with R(f) values 0.42 ± 0.03 and 0.61 ± 0.03 for kutkoside and picroside-I, respectively. The method was validated in terms of specificity, linearity, accuracy and precision. RESULTS: The content of kutkoside and picroside-I was found to be 2.18 and 1.90%, respectively, and was comparable with those obtained by HPLC. The linearity was found to be in the range of 80-480 ng/spot for both kutkoside and picroside-I. The average recovery values were found to be 96.5 and 96.0% for kutkoside and picroside-I, respectively. CONCLUSION: The developed method was found to be relatively simple, precise and reproducible for the simultaneous quantitation of kutkoside and picroside-I. The method does not employ any derivatisation procedure and can be used as a quality control tool for the routine analysis of commercial Kutki extracts.

Use of hearing protection on military operations.

Noise Induced Hearing Loss (NIHL) continues to be a significant source of morbidity for UK service personnel. The Personal Interfaced Hearing Protection (PIHP) system was procured as an Urgent Operational Requirement primarily to provide a connection to the Personal Role Radio. Through its custom- mould design it was also expected to act as Personal Protective Equipment (PPE) against NIHL in a method that had not been possible previously. The aim of this paper was to evaluate user opinion of the four current methods of hearing protection including PIHP. 108 service personnel serving in a Forward Operating Base in Afghanistan were questioned anonymously using a paper-based questionnaire. The response rate was 78%, reflecting the difficulties in anonymous data collection in an environment such as this. PIHP was perceived to be more practical for use on the ranges or patrolling than the standard methods of hearing protection. Although only 4% were routinely wearing PIHP on patrol, this was related to understandable reluctance in wearing a new system that had only been issued immediately prior to this deployment. Early design problems highlighted by our study, such as changes in the design of the ear-piece to improve retention, have already been implemented. Compliance will be improved through a dedicated educational programme to explain its relevance. Despite the logistical difficulties in providing custom- made ear moulds, the PIHP system must be delivered to all service personnel early in their career so they are confident in its use long before operational deployment. Future research is required to ascertain the effectiveness of PIHP as a method of PPE in reducing the incidence of NIHL.

Gene ontology enrichment analysis of PPAR-γ modulators from Cassia glauca in diabetes mellitus.

BACKGROUND: PPAR-γ has an integrative role in the management of insulin resistance; ligands of this receptor have emerged as potent insulin sensitizers and may modulate proteins involved in the pathogenesis of diabetes mellitus. Hence the present study is aimed to identify PPAR-γ modulators from the plant Cassia glauca and predict the ontology enrichment analysis utilizing various in-silico tools. METHODS: ChEBI database was used to mine the phytoconstituents present in the plant C. glauca, SwissTargetPrediction database was used to identify the targets, and scrutinizing of phytoconstituents modulating PPAR-γ was performed. Autodock4.0 was used to dock phytoconstituent ligands with the target PPAR-γ. Multiple open-source databases and in-silico tools were utilized to predict the drug-likeness characters and predict side effects of the phytoconstituents modulating PPAR-γ and STRING database was used to construct a network between the modulated genes. RESULTS: Twenty-four phytoconstituents were identified from the plant Cassia glauca from which four were found to modulate PPAR-γ, sennoside was predicted to have the greatest drug-likeness score and a significantly less side effect whereas diphenyl sulfone was predicted to show hepatotoxicity with the greatest pharmacological activity of 0.815. [epicatechin-(4beta- > 8)]5-epicatechin showed the lowest binding affinity with target PPAR-γ i.e. -8.6 kcal/mol and possessing a positive drug-likeness score with no side effect data. CONCLUSION: Bioctives were found free from probable side effects leaving out diphenyl sulfone having a prediction of hepatotoxicity, the anti-diabetic property of the plant may be due to the presence of [epicatechin-(4beta- > 8)]5-epicatechin which needs further validation by in-vitro and in-vivo protocols.

Development and validation of a method for densitometric analysis of berberine in herbal extract and polyherbal formulation.

OBJECTIVE: To develop a new high-performance thin-layer chromatography (HPTLC) method for the quantification of berberine in herbal extract and pharmaceutical dosage form. MATERIALS AND METHODS: The HPTLC was performed on aluminium foil plates coated with 200 µm silica gel 60F(254). Linear ascending development with toluene:ethyl acetate:formic acid:methanol 9:9:3:1 (v/v/v/v) was performed at room temperature (25 ± 2°C) in a twin-trough glass chamber saturated with mobile-phase vapour. Compact bands (R(F) 0.58 ± 0.02) were obtained for berberine. Spectrodensitometric scanning was performed in fluorescence mode at 350 nm. The method was validated for precision, recovery, robustness, specificity, and detection and quantification limits, in accordance with International Conference on Harmonization guidelines. RESULTS: Linear regression analysis of the calibration plots showed a good linear relationship (r(2) = 0.9996 ± 0.0001) between peak area and concentration in the range 10-100 ng/band, respectively. The limits of detection and quantification were 2.8 and 9.3 ng/band. The recovery of the method was 98.5-100.6%. CONCLUSION: The above method was a rapid and cost-effective quality-control tool for routine analysis of berberine in herbal extracts and in pharmaceutical dosage form.

Gene set enrichment analysis of α-amylase and α-glucosidase inhibitors of Cassia glauca.

BACKGROUND: The present study aimed to evaluate in vitro α-amylase and α-glucosidase inhibitory activity of various extracts of Cassia glauca, predict the binding affinity of multiple phytoconstituents with both enzymes via in silico molecular docking and identify the probably modulated pathways by the lead hit. METHODS: Different extracts of Cassia glauca i.e. acetone, ethanol, and aqueous extracts were evaluated for α-amylase and α-glucosidase inhibitory activity using in vitro method in which starch and 4-Nitrophenyl ß-D-glucopyranoside were used as substrate respectively. Similarly, the docking study was performed using autodock4 to predict the binding affinity of phytoconstituents with α-amylase and α-glucosidase. After docking, ten different poses were obtained for the ligand molecule. Among them, the pose of ligand molecule with the lowest binding energy was visualized in Discovery Studio 2019. RESULTS AND CONCLUSION: Among the multiple extracts, the aqueous extract showed the highest α-amylase (IC50:652.10 ± 20.09) and α-glucosidase (IC50:482.46 ± 8.70) inhibitory activity. Similarly, cassiaoccidentalin B was predicted to have the highest binding affinity with both enzymes. The potency of aqueous extract to inhibit α-amylase and α-glucosidase could be due to multiple water-soluble compounds like saponins, flavonoids, and glycosides.