Characterization of bacteriocin produced by Brevibacillus laterosporus Tk3 isolated from 'tatwakhar' - a flour prepared from seeds of Aesculus indica in remote areas of Himachal Pradesh India.

Bacteriocin producing strain Brevibacillus laterosporus TK3, was isolated from 'Tatwakhar'- a flour prepared from seeds of Indian Horse Chestnut (Aesculus indica). Bacterial strain Brevibacillus laterosporus TK3 identified by morphological, biochemical techniques followed by 16S rRNA gene sequencing. The 16S rRNA sequence of bacteriocin producer was deposited in NCBI GenBank under accession no. KP861913.1. Bacteriocin of Brevibacillus laterosporus TK3 showed strong antagonistic activity against food spoiling/pathogenic bacteria viz. Listeria monocytogenes, Staphylococcus aureus and Clostridium perfringens. Bacteriocin production by Brevibacillus laterosporus TK3 was enhanced by optimizing production time, pH of the medium, inoculum size and incubation temperature. Maximum bacteriocin activity (6000 AU/ml) was recorded/obtained in basal salt medium of pH 5.5 with an inoculum size of 1.5 OD at 10% and incubation period of 24h at 35 °C. The bacteriocin was purified by single step gel exclusion chromatography. Molecular weight of active bacteriocin from Brevibacillus laterosporous TK3 was found to be 6 kDa according to SDS PAGE. The molecular mass of purified bacteriocin was confirmed as 5953.89 Da by MALDI TOF analysis. The purified bacteriocin was found desirable/suitable for food preservation as it showed wide spectrum of antimicrobial activity, resistance to high temperature, wide pH range and sensitivity to proteolytic enzymes thus, making it safe for human consumption.

Patch testing with expired Indian patch test kits: Results of a pilot study.

BACKGROUND: The reliability of patch testing with expired Indian standard patch test kits has been not evaluated before. METHODS: Thirty adults (men:women 25:5) with allergic contact dermatitis were divided into three groups of ten patients each for patch testing by Finn chamber® method using Indian standard patch test kits having expiry in 2016, 2015 and 2014. The results were compared with those from a new kit with 2018 expiry. RESULTS: Ten patients in group-1, eight patients in group-2 and seven patients in group-3 developed positive reactions of identical intensities and mostly from identical allergens from all four kits. The major contact allergens eliciting positive reactions of identical intensities were parthenium in nine, five and three patients, colophony in four, one and zero patients, fragrance mix in three, three and one patients, thiuram mix in three, one and one patients, and paraphenylene diamine in two, one and three patients from group-1,-2, and -3, respectively. LIMITATIONS: Small number of patients in each group remains the major limitation of the study. Whether or not these results can be extrapolated with patch test results from other similar patch test kits available across countries also needs confirmation. CONCLUSION: The patch test allergens can be used beyond labeled expiry dates but needs confirmation by a few large studies and using other available patch test kits. This is important as the relevance of patch test results for individual allergen in this scenario may remain debatable requiring careful interpretation.

Clinical, Occupational and Allergological Profile of 455 Patients with Occupational Contact Dermatitis: A 5-Year Study from a Tertiary Care Center of North India.

OBJECTIVES: To evaluate clinical and occupational profile and common allergens in patients with occupational contact dermatitis (OCD). MATERIALS AND METHODS: The records of 455 (M:F 2:1) patients aged 18-85 years with allergic contact dermatitis were analyzed retrospectively. The diagnosis of OCD and patterns of dermatitis were defined by standard criteria. Indian standard series comprising 20 allergens and when suspected patient's own products were patch tested by Finn chamber method as per European Society of Contact Dermatitis guidelines and relevance of positive results was defined clinically. RESULTS: Airborne contact dermatitis (27.7%), acral dermatitis (14.1%), hand dermatitis (12.9%), acrofacial dermatitis (12.7%), and facial dermatitis (10.5%) were the common patterns. Agriculturists (51.2%), homemakers (27.9%), office workers (24.6%), and construction workers (4.6%) comprised the majority. Positive patch test results in 58% cases were from parthenium (31.7%), p-paraphenylenediamine (PPD) (22.9%), nickel (16%), fragrance mix (11%), potassium dichromate (10.7%), cobalt (7.6%), and mercaptobenzothiazole (4.9%). Hair colorants, shoe chips, and shaving cream also produced relevant positive reactions. Parthenium, PPD, fragrance mix, and potassium dichromate in agriculturists; nickel, parthenium, PPD, fragrance mix, and potassium dichromate in women, and potassium dichromate and parthenium in construction workers elicited the most positive reactions. PPD and hair colorants elicited positive reaction mainly in office workers. CONCLUSIONS: The agriculturists, homemakers, and construction workers have OCD most frequently. Parthenium in farmers, potassium dichromate in construction workers, nickel in women, and PPD in office workers were the major contact allergens. The study is limited by its retrospective design, small number of patients, and limited number of patch test allergens.

Clinico-investigative attributes of 122 patients with hirsutism: A 5-year retrospective study from India.

BACKGROUND: Hirsutism is common across ethnicities and a significant cause of negative self-esteem from presumptive loss of femininity. It remains understudied in Indian patients. OBJECTIVE: We studied the clinical and investigative attributes of patients with hirsutism. METHODS: The medical records of 233 patients with hirsutism diagnosed between 2014 and 2019 were analyzed retrospectively. RESULTS: The complete records of 122 patients age 14 to 45 years were available. Approximately 32% were adolescents, and 50% patients were age 21 to 30 years. The mean ±â€¯standard deviation modified Ferriman-Gallway (mF-G) score was 17.95 ±â€¯10.58, and hirsutism was graded zero/mild in 57.4% patients. Polycystic ovaries were present in 29.5% of patients. Serum-free testosterone levels were elevated in 16.4% of patients. Associated signs of clinical hyperandrogenism, such as acne, obesity, acanthosis nigricans, and menstrual irregularities, were present in 12 patients (9.8%). Familial hirsutism occurred in 13% of patients and was idiopathic in 10.7% of patients. Significantly more patients (47.7% vs. 27.9%) with severe hirsutism did not complete investigations compared with those with mild hirsutism (45.9% vs. 21.6%), which may be due to the high cost of investigative work-up (as often stated by many patients) compared with periodic depilation. LIMITATIONS: The mF-G score is a visual and subjective scale, and its validity remains limited by interobserver variations. The score's cut-off values may vary among races/ethnicities. A small number of patients and the retrospective study design are other limitations. CONCLUSION: Hirsutism of polycystic ovary syndrome or idiopathic origin is not uncommon in Indian women. Hirsutism of adrenal or thyroid origin remains uncommon. Self-referral and high cosmetic concerns for facial hair were common in adolescents and young unmarried patients, and the majority had an mF-G score of < 8 to 16. Cut-off values for the mF-G score specific to our population assigning higher than current mF-G value to facial hair are highly desirable.

Adults with a more extensive body involvement, moderate to extremely severe vitiligo and a prolonged clinical course have an early onset in childhood in addition to other prognostic factors as compared to individuals with later-onset vitiligo.

BACKGROUND: The extent and disease severity, duration and other associated prognostic cofactors in vitiligo in adults may vary with the age of onset (before or after 10 years of age). OBJECTIVES: To compare extent and disease severity, duration and other cofactors in adults with early-onset and late-onset vitiligo. METHODS: The medical records of 408 (M:F 1:1.1) adults aged 20-75 years diagnosed with vitiligo between January 2016 and December 2019 were examined retrospectively. The extent and severity of vitiligo were defined. Characteristics of vitiligo with early onset and late onset were compared statistically and odds ratios calculated for risk assessment. RESULTS: 31 (7.6%, M:F 1:2.4) patients had early-onset vitiligo, and 377 (92.4%, M:F 0.8:1) patients had later-onset vitiligo. Compared to late onset, patients with early-onset vitiligo had a significant number of males (71% vs 45.9%), higher percentages of body surface area involvement and moderate to extremely severe disease (29% vs 10.6%), longer duration of disease (41.9% vs 9%), Koebner's phenomenon (48.4% vs 15.6%) and halo nevus (9.7% vs 1.9%). Differences between the two groups were not significant for types of vitiligo, family history of vitiligo and presence of cutaneous and systemic/autoimmune diseases. CONCLUSION: The adults, males in particular, with generalised vitiligo (>10% BSA involvement) appear to have an early onset and a prolonged clinical course. The presence of Koebner's phenomenon and halo nevus in patients with early-onset vitiligo was other poor prognostic factors compared to patients with late-onset vitiligo. The retrospective, hospital-based cross-sectional design and small sample size for stratification remain major limitations.

Characteristics of Vitiligo in Children and Adolescents.

Vitiligo in children and adolescents displays some distinct features, which may affect its clinical course, therapeutic outcome, and prognosis. We studied 579 children and adolescents with vitiligo, comprising 275 (47.5%) boys and 304 (52.5%) girls (male:female ratio [m:f], 1:1.1) aged between 2 and 19 years (mean ± SD 11.13 ± 4.23 years). The majority of children (301, 52%) were aged >5-12 years, and 221 (38.2%) were adolescents; onset of vitiligo in the above groups occurred between the ages of 2 and 19 years (mean ± SD 9.18 ± 4.08 years). The majority of patients (337, 58.2%) had developed vitiligo between 5 and 12 years of age, and 332 (57.4%) patients had a medical consultation within 1 year of the onset of the disease. The involvement of up to 10% of body surface area in 569 (98.3%) patients, generalized vitiligo in 328 (56.7%) patients, and focal vitiligo in 158 (27.3%) patients were the major presentations. Only 150 (15.9%) patients had family members affected by vitiligo, and associated systemic disorders, predominately thyroid abnormalities, occurred in three (0.5%) patients. Vitiligo was more frequent in children aged 5-12 years, and it affected girls with a slight preponderance, commonly presenting as generalized vitiligo and focal/localized vitiligo. Patients with a family history of vitiligo had an earlier onset but without a statistically significant difference. Local trauma is an important trigger, and screening for thyroid disorders appears important.

Skin photo-protection with phytochemicals against photo-oxidative stress, photo-carcinogenesis, signal transduction pathways and extracellular matrix remodeling-An overview.

Excessive exposure of skin to UV radiation triggers the generation of oxidative stress, inflammation, immunosuppression, apoptosis, matrix-metalloproteases production, and DNA mutations leading to the onset of photo ageing and photo-carcinogenesis. At the molecular level, these changes occur via activation of several protein kinases as well as transcription pathways, formation of reactive oxygen species, and release of cytokines, interleukins and prostaglandins together. Current therapies available on the market only provide limited solutions and exhibit several side effects. The present paper provides insight into scientific studies that have elucidated the positive role of phytochemicals in counteracting the UV-induced depletion of antioxidant enzymes, increased lipid peroxidation, inflammation, DNA mutations, increased senescence, dysfunctional apoptosis and immune suppression. The contribution of phytochemicals to the downregulation of expression of oxidative-stress sensitive transcription factors (Nrf2, NF-Kb, AP-1 and p53) and protein kinases (MSK, ERK, JNK, p38 MAPK, p90RSK2 and CaMKs) involved in inflammation, apoptosis, immune suppression, extracellular matrix remodelling, senescence, photo ageing and photo-carcinogenesis, is also discussed. Conclusively, several phytochemicals hold potential for the development of a viable solution against UV irradiation-mediated photo ageing, photo-carcinogenesis and related manifestations.

Pentacyclic triterpenes: New tools to fight metabolic syndrome.

BACKGROUND: Metabolic syndrome is a combination of dysregulated cardiometabolic risk factors characterized by dyslipidemia, impaired glucose tolerance, insulin resistance, inflammation, obesity as well as hypertension. These factors are tied to the increased risk for type-II diabetes and cardiovascular diseases including myocardial infarction in patients with metabolic syndrome. PURPOSE: To review the proposed molecular mechanisms of pentacyclic triterpenes for their potential use in the metabolic syndrome. METHODS: PubMed, Science Direct, and Google Scholar database were searched from commencement to April 2018. Following keywords were searched in the databases with varying combinations: "metabolic syndrome", "pentacyclic triterpenes", "transcription factors", "protein kinase", "lipogenesis", "adipogenesis", "lipolysis", "fatty acids", "gluconeogenesis", "cardiovascular", "mitochondria", "oxidative stress", "pancreas", "hepatic cells", "skeletal muscle", "3T3-L1", "C2C12", "obesity", "inflammation", "insulin resistance", "glucose uptake", "clinical studies" and "bioavailability". RESULTS: Pentacyclic triterpenes, such as asiatic acid, ursolic acid, oleanolic acid, 18ß-glycyrrhetinic acid, α,ß-amyrin, celastrol, carbenoxolone, corosolic acid, maslinic acid, bardoxolone methyl and lupeol downregulate several metabolic syndrome components by regulating transcription factors, protein kinases and enzyme involved in the adipogenesis, lipolysis, fatty acid oxidation, insulin resistance, mitochondria biogenesis, gluconeogenesis, oxidative stress and inflammation. CONCLUSION: In vitro and in vivo studies suggests that pentacyclic triterpenes effectively downregulate various factors related to metabolic syndrome. These phytochemicals may serve as promising candidates for clinical trials for the management of metabolic syndrome.

Natural AMPK Activators: An Alternative Approach for the Treatment and Management of Metabolic Syndrome.

This review covers recent discoveries of phytoconstituents, herbal extracts and some semi-synthetic compounds for treating metabolic syndrome with AMPK activation as one of their mechanisms of action. Recent researches have demonstrated AMPK activation to ameliorate multiple components of metabolic syndrome by regulating a balance between anabolic and catabolic cellular reactions. The review attempts to delineate the AMPK activation by natural agents from the perspective of its functional consequences on enzymes, transcription factors and signaling molecules and also on other potential factors contributing in the amelioration of metabolic syndrome.

Synthesis of novel inhibitors of α-amylase based on the thiazolidine-4-one skeleton containing a pyrazole moiety and their configurational studies.

Postprandial hyperglycemia can be controlled by delaying the absorption of glucose resulting from carbohydrate digestion. α-Amylase is the initiator of the hydrolysis of polysaccharides, and therefore developing α-amylase inhibitors can lead to development of new treatments for metabolic disorders like diabetes mellitus. In the present work, we set out to rationally develop α-amylase inhibitors based on the thiazolidine-4-one scaffold. The structures of all these newly synthesized hybrids were confirmed by spectroscopic analysis (IR, 1H-NMR, MS). The appearance of two sets of signals for some protons in 1H NMR revealed the existence of a mixture of 2E,5Z (37.1-42.0%) and 2Z,5Z isomers (58.4-62.8%), which was further supported by DFT studies. All the newly synthesized compounds have potential inhibitory properties as revealed through in vitro α-amylase inhibition activity. Compound 5a at 100 µg mL-1 concentration showed a remarkable inhibition of 90.04%. In vitro α-amylase inhibition was further supported by docking studies of compound 5a against the active site of human pancreatic α-amylase (PDB ID: ; 2QV4). The docking studies revealed that the bonding interactions found between 5a and human pancreatic α-amylase are similar to those responsible for α-amylase inhibition by acarbose.

Neuropharmacological activities of Taxus wallichiana bark in Swiss albino mice.

AIMS: The bark of Taxus wallichiana is widely used for preparing a decoction and consumed as a tea by several tribal communities of the Indian subcontinent. The sedative, motor coordination, anxiolytic, and antidepressant effects of the hydroalcoholic extract of T. wallichiana bark and its ethylacetate fraction were evaluated in mice models of behavior analysis. MATERIALS AND METHODS: The effects were evaluated on diazepam-induced sleeping time, elevated plus maze and light and dark box, and on the forced swimming test. General locomotor activity and motor coordination effects were evaluated in the actophotmeter and rota-rod tests respectively. STATISTICAL ANALYSIS: Results are expressed as mean ± standard error of the mean. Statistical analysis was performed using ANOVA, followed by post-hoc Dunnett's test. *P < 0.05, **P < 0.01, ***P < 0.001 were considered as significant. RESULTS: Both the hydroalcoholic extract and ethylacetate fraction showed a marked decrease in latency of sleep onset, prolonged the diazepam-induced sleeping time, decreased spontaneous locomotor activity; whereas ethylacetate fraction produced anxiolytic and antidepressant activity. CONCLUSIONS: Both hydroalcoholic extract and its ethylacetate fraction of the bark of T. wallichiana have bioactive principles, which induce neuropharmacological changes.

A review of traditional use, phytoconstituents and biological activities of Himalayan yew, Taxus wallichiana.

Plants synthesize certain phytoconstituents for their protection, which, because they are not of primary need, are known as secondary metabolites. These secondary metabolites of plants, have often been found to have medicinal uses for human beings. One such gymnosperm having secondary metabolites of medicinal potential for humans is Taxus wallichiana (Himalayan yew). Besides being the source of taxol, this plant has been investigated for its essential oil, diterpenoids, lignans, steroids, sterols and biflavonoids. Traditionally, it is used to treat disorders of the digestive, respiratory, nervous and skeletal systems. Although pharmacologically underexplored, it has been used for antiepileptic, anti-inflammatory, anticancer, antipyretic, analgesic, immunomodulatory and antimicrobial activities. The present review compiles traditional uses, phytochemical constituents (specifically the secondary metabolites) pharmacological activities and the toxicity of T. wallichiana.

Cardioprotective effect of ammonium glycyrrhizinate against doxorubicin-induced cardiomyopathy in experimental animals.

OBJECTIVE: The objective of this study was to evaluate the cardioprotective effect of herbal bioactive compound ammonium glycyrrhizinate against doxorubicin-induced cardiomyopathy, in experimental animals. MATERIALS AND METHODS: Ammonium glycyrrhizinate (50, 100, 200 mg/kg, p.o.) was administered for four weeks in albino rats. Cardiomyopathy was induced with a dose of 2.5 mg/kg i.p. of doxorubicin on 1(th), 7(th), 14(th), 21(th), 28(th) day in the experimental animals. At the end of the experiment, on 29(th) day, serum and heart tissues were collected and hemodynamic, biochemical and histopathological studies were carried out. RESULTS: Administration of doxorubicin in normal rats showed significant (P < 0.001) changes in body weight, feed intake, urine output, hemodynamic parameters like (blood pressure, heart rate, cardiac output) and in lipid profile (cholesterol, triglyceride, high density lipoprotein, low density lipoprotein, very low density lipoprotein) indicating cardiomyopathy symptoms. Animals treated with ammonium glycyrrhizinate significantly (P < 0.05) decreased triglyceride, cholesterol, low density lipoprotein (LDL) and very low density lipoprotein (VLDL) levels. Moreover, high density lipoprotein (HDL) levels increased in rats treated with ammonium glycyrrhizinate as compared with the normal group. CONCLUSION: Ammonium glycyrrhizinate is effective in controlling serum lipid profile and cardiac complications in experimentally induced cardiomyopathy in animals.