T-2 toxin induces dermal inflammation and toxicity in mice: The healing potential of menthol.

According to the World Health Organization and the Food and Agricultural Organization of the United Nations, T-2 is one of the most harmful food-toxic chemicals, penetrates intact skin. The current study examined the protective benefits of menthol topical treatment on T-2 toxin-induced cutaneous toxicity in mice. Lesions were observed on the skin of the T-2 toxin-treated groups at 72 and 120 h. The T-2 toxin (2.97 mg/kg/bw)-treated group developed skin lesions, skin inflammation, erythema, and necrosis of skin tissue in contrast to the control group. Our findings reveal that topical application of 0.25% and 0.5% MN treated groups resulted in no erythema or inflammation, and normal skin was observed with growing hairs. The 0.5% MN administered group demonstrated an 80% blister and erythema healing effect in in vitro tests. In addition, MN dose-dependently suppressed ROS and lipid peroxidation mediated by the T-2 toxin up to 120%. Histology discoveries and the immunoblotting investigations with the downregulation of i-NOS gene expression confirmed the validity of menthol activity. Further molecular docking experiments of menthol against the i-NOS protein demonstrated stable binding efficacy with conventional hydrogen bond interactions, indicating compelling evidence of menthol's anti-inflammatory effects on the T-2 toxin-induced skin inflammation.

Salivary Menopausal Markers and Oral Health Status - A Hidden Hook Up.

Aim and Objectives: The aim of our study was to evaluate salivary estradiol and salivary calcium in postmenopausal women with varying degrees of oral dryness. The primary objective was to establish the interrelationship between salivary parameters and oral health status among menopausal women and compare the same with premenopausal women and normal controls. Materials and Methods: The study included 60 women Group I consisted of healthy menstruating women between 25 and 34 years of age. In Group II premenopausal women between 35 and 45 years of age were present and Group III consisted of menopausal women between 45 and 60 years of age. Unstimulated saliva was collected from the participants and estradiol analysis was done using ELISA method and calcium analysis was done using Arsenazo III reaction using colorimetric method. The oral health status in these patients was determined by using xerostomia score, Russell's periodontal score, and oral hygiene index. The values obtained were subjected to statistical analysis and the results were derived. Results: On oral examination, most of them had poor oral hygiene, periodontal disease, and moderate to severe levels of xerostomia. Salivary estradiol levels were low and salivary calcium levels were high among postmenopausal women and as salivary estradiol levels decreased there was an increase in xerostomia scores and salivary calcium. And also as salivary calcium levels increased the periodontal disease scores increased. All parameters were within normal limits among healthy menstruating women. Conclusion: Saliva can be a preferred medium and an emerging alternative for serum to estimate estradiol and calcium levels. As a dentist, we have to educate them about the oral changes they will experience during menopause and emphasize its strong association between low estradiol levels. Oral hygiene instructions should be given for the maintenance of healthy periodontium. Menopausal women who experience severe postmenopausal symptoms can be identified and the dentist and gynecologist can work hand in hand to treat the symptoms of these women.

Identification of potential biomarkers for lung adenocarcinoma.

Lung adenocarcinoma (LUAD) is the most predominant subtype of lung cancers and is one of the leading causes of cancer related mortality worldwide. Despite the advancements in the field of cancer diagnostics and therapeutics, detection at an early stage using reliable biomarkers is an unmet clinical need for a plethora of cancers, including LUAD, thus attributing to poor prognosis. In view of this, to identify potential biomarkers and therapeutic candidate genes, the expression of all known human genes was screened in the publicly available 'The Cancer Genome Atlas' (TCGA) samples of LUAD patients which resulted in the identification of overexpressed genes. Further analysis of these genes across various patient sample datasets revealed that ZNF687, ODR4, PBXIP1, PYGO2, METTL3, PIGM and RAD1 are consistently more highly expressed in LUAD. Higher expression of these genes either alone or in combination is correlated with poor survival of LUAD patients. Hence, in this study we propose that these identified genes could serve as potential candidates as gene signatures or biomarkers for LUAD that require further investigation in large cohorts of LUAD samples.

Chemical composition, antioxidant potential, macromolecule damage and neuroprotective activity of Convolvulus pluricaulis.

Herbal medicines are known to mitigate radical induced cell damage. Hence identification and scientific validation of herbal medicines contribute to better use in Ayurvedic/Unani research. In the present study, we investigated antioxidant and anti-apoptotic properties of Convolvulus pluricaulis (C. pluricaulis). C. pluricaulis exhibited antioxidant potential evident by free radical scavenging activities. C. pluricaulis pretreatment inhibited H2O2 induced macromolecule damage such as plasmid DNA damage and AAPH induced oxidation of bovine serum albumin and lipid peroxidation of rat hepatic tissues. Further to identify the neuroprotective properties of C. pluricaulis, SHSY5Y cells were treated with H2O2 with or without pretreatment of C. pluricaulis. The C. pluricaulis pretreatment at 50 µg/ml dose exhibited 50% cell survival against 100 µM H2O2 challenge for 24 h and it also decreased the lactate dehydrogenase leakage. Further C. pluricaulis pretreatment restored and regulated the antioxidant and apoptosis markers such as SOD, CAT, p53, and caspase-3 and inhibited, reactive oxygen species generation and depolarization of the mitochondrial membrane. C. pluricaulis possess a high content of flavonoids and polyphenols and GC-MS and FTIR analysis showed a wide variety of compounds which may contribute to the observed effects.

Anti-inflammatory and anti-oxidant effects of Cardamom (Elettaria repens (Sonn.) Baill) and its phytochemical analysis by 4D GCXGC TOF-MS.

Elleteria repens is a large cardamom used in the culinary preparations. In the present study, we have evaluated in vitro antioxidant and free radical scavenging activities E. repens hexane extract (ERH) exhibited DPPH and metal chelating activity with IC50 values of 464±28.3µg/ml, 199±7.2µg/ml whereas the reducing power and antioxidant activities are found to be 289±14.6 AAE/mg, 468±22.7 GAE/mg. The observed antioxidant activities could be correlated with metabolites such as polyphenol, flavonoid, and terpenoid group of compounds identified in hexane fraction of E. repens by 4D GCXGC TOF-MS. Further ERH was evaluated for its protective properties against macromolecules such as DNA, protein and lipid damage. The extract showed protection against H2O2 induced DNA damage and inhibited AAPH induced protein oxidation and lipid peroxidation. Moreover, ERH administration to rats at 50 and 100mg/kg inhibited carrageenan-induced paw edema, and down-regulated cytokines such as COX-2, IL-6, and TNF-α and inhibited i-NOS mediated NO generation. E. repens also exhibited antioxidant effects by restoring SOD, catalase, GSH levels and inhibited lipid peroxidation in carrageenan challenged rats. Overall, the results suggest that E. repens may be useful in combating inflammation and oxidative stress.

Growth Inhibition and Morphological Alteration of Fusarium sporotrichioides by Mentha piperita Essential Oil.

OBJECTIVE: The aim of this study is to determine the phytochemical composition, antifungal activity of Mentha piperita essential oil (MPE) against Fusarium sporotrichioides. METHODS: The phytochemical composition was conducted by gas chromatography mass spectrometry (GC MS) analysis and mycelia growth inhibition was determined by minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC), the morphological characterization was observed by scanning electron microscopy. Finally, the membrane permeability was determined by the release of extracellular constituents, pH, and total lipid content. RESULT: In GC MS analysis, 22 metabolites were identified such as menthol, l menthone, pulegone, piperitone, caryophyllene, menthol acetate, etc. The antifungal activity against targeted pathogen, with MIC and MFC 500 µg/mL and 1000 µg/mL, respectively. The MPE altered the morphology of F. sporotrichoides hyphae with the loss of cytoplasm content and contorted the mycelia. The increasing concentration of MPE showed increase in membrane permeability of F. sporotrichoides as evidenced by the release of extracellular constituents and pH with the disruption of cell membrane indicating decrease in lipid content of F. sporotrichoides. CONCLUSION: The observed results showed that MPE exhibited promising new antifungal agent against Fusarium sporotrichioides. SUMMARY: F. sporotrichioides, filamentous fungi contaminate to corn and corn--based productsF. sporotrichioides mainly responsible for the production of T-2 toxinPhytochemical composition was conducted by gas chromatography--mass spectrometry analysisMentha piperita essential oil (MPE) is commonly known as peppermintThe F. sporotrichioides growth was inhibited by MPE (minimum inhibitory concentration, minimum fungicidal concentration)Morphological observation by scanning electron microscope. Abbreviations Used: Cfu: Colony forming unit; DMSO: Dimethyl sulfoxide, °C: Degree celsius; F. Sporotrichoides: Fusarium sporotrichioides; EOs: Essential oils; M: Molar, g: Gram/gravity, mg: Milligram; µg: Microgram, ml: Milliliter; mm: Millimeter, min: Minutes; M. piperita: Mentha piperita, MIC: Minimum inhibitory concentration; MFC: Minimum fungicidal concentration; MAE: Mentha arvensis essential oil; Na2SO4: Sodium sulfate; pH: Potential Hydrogen; PDB: Potato Dextrose Broth; SEM: Scanning electron microscope.

LC-ESI-MS/MS analysis of total oligomeric flavonoid fraction of Cyperus rotundus and its antioxidant, macromolecule damage protective and antihemolytic effects.

In the present investigation, we identified the phytochemical constituents of total oligomeric flavonoid fraction (TOF) of Cyperus rotundus by LC-ESI-MS/MS and also demonstrated its antihemolytic effects against 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) induced hemolysis of rat erythrocytes. Our results of TOF extract exhibited DPPH, metal chelating, ABTS, NO and hydroxyl radical scavenging activities with an IC50 values of 23.72±1.6, 52.45±2.88, 9.8±0.42, 6.5±0.33 and 120±6.83µg/ml respectively, whereas total antioxidant and reducing power activities were 194±12.5µg GAE/mg extract and 145±8.3µg AAE/mg extract. The extract showed potent inhibitory activity against AAPH induced plasmid DNA damage, protein oxidation and lipid peroxidation. The TOF extract mitigates AAPH induced hemolysis and exhibits ∼50% antihemolytic activity. TOF pretreatment also preserved morphology of erythrocytes as observed and measured by light microscope and atomic force microscope analysis. Furthermore, the TOF fraction effectively inhibited AAPH induced LDH release, ROS generation and lipid peroxidation. Taken together, our data demonstrate the antihemolytic activity of C. rotundus against AAPH induced oxidative stress of erythrocytes, and was associated with the decrease in oxidative stress, cellular damage and protection of macromolecules. In conclusion, the effects might be correlated with high content of flavonoids and polyphenols identified in C. rotundus. This suggests the clinical application of TOF fraction of C. rotundus against ROS induced cell death.

Celastrus paniculatus Willd. mitigates t-BHP induced oxidative and apoptotic damage in C2C12 murine muscle cells.

Identification, exploration and scientific validation of antioxidant rich herbal extracts to mitigate the radical induced cell damage provide new insights in the field of ayurvedic research/therapies. In the present study, we evaluated the anti-oxidant and anti-apoptotic potential of Celastrus paniculatus seed extract (CPSE) against tertiary butyl hydroperoxide (t-BHP) induced mice muscle cell damage. The extract at a dose of 50 µg/ml protected the cells up to 70 % as evidenced by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell survival assay and also prevented LDH leakage against t-BHP induced cytotoxicity. CPSE showed potential antioxidant activity by restoring mitochondrial membrane potential and inhibited reactive oxygen species generation and lipid peroxidation. CPSE pretreatment also regulated the antioxidant markers such as superoxide dismutase and catalase enzymes content and proteins expression. Further CPSE showed anti-apoptotic effects by regulating cytochrome-C and heat shock protein-70 expression and also showed 43 % muscle cell DNA damage inhibitory activity against t-BHP challenge as observed by single cell gel electrophoresis assay. Overall the extract inhibits the muscle cell damage, thus explaining the possible anti-oxidant/anti-apoptotic defense status of the C. paniculatus seed extract.

Structural basis for the energetics of jacalin-sugar interactions: promiscuity versus specificity.

Jacalin, a tetrameric lectin, is one of the two lectins present in jackfruit (Artocarpus integrifolia) seeds. Its crystal structure revealed, for the first time, the occurrence of the beta-prism I fold in lectins. The structure led to the elucidation of the crucial role of a new N terminus generated by post-translational proteolysis for the lectin's specificity for galactose. Subsequent X-ray studies on other carbohydrate complexes showed that the extended binding site of jacalin consisted of, in addition to the primary binding site, a hydrophobic secondary site A composed of aromatic residues and a secondary site B involved mainly in water-bridges. A recent investigation involving surface plasmon resonance and the X-ray analysis of a methyl-alpha-mannose complex, had led to a suggestion of promiscuity in the lectin's sugar specificity. To explore this suggestion further, detailed isothermal titration calorimetric studies on the interaction of galactose (Gal), mannose (Man), glucose (Glc), Me-alpha-Gal, Me-alpha-Man, Me-alpha-Glc and other mono- and oligosaccharides of biological relevance and crystallographic studies on the jacalin-Me-alpha-Glc complex and a new form of the jacalin-Me-alpha-Man complex, have been carried out. The binding affinity of Me-alpha-Man is 20 times weaker than that of Me-alpha-Gal. The corresponding number is 27, when the binding affinities of Gal and Me-alpha-Gal, and those of Man and Me-alpha-Man are compared. Glucose (Glc) shows no measurable binding, while the binding affinity of Me-alpha-Glc is slightly less than that of Me-alpha-Man. The available crystal structures of jacalin-sugar complexes provide a convincing explanation for the energetics of binding in terms of interactions at the primary binding site and secondary site A. The other sugars used in calorimetric studies show no detectable binding to jacalin. These results and other available evidence suggest that jacalin is specific to O-glycans and its affinity to N-glycans is extremely weak or non-existent and therefore of limited value in processes involving biological recognition.

Design, synthesis, and evaluation of 3,4-dihydro-2H-[1,4]diazepino[6,7,1-hi]indol-1-ones as inhibitors of poly(ADP-ribose) polymerase.

The design, synthesis, and biological evaluation of potent inhibitors of poly(ADP-ribose) polymerase-1 (PARP-1) are reported. A novel series of 3,4-dihydro-2H-[1,4]diazepino[6,7,1-hi]indol-1-ones were designed using a combination of protein structure-based drug design, molecular modeling, and structure-activity relationships (SAR). These novel submicromolar inhibitors possess a tricyclic ring system conformationally restricting the benzamide in the preferred cis orientation. The compounds were designed to optimize space-filling and atomic interactions within the NAD+ binding site of PARP-1. Previously described and newly adapted methods were applied to syntheses of these tricyclic inhibitors. Various modifications were made to the diazepinoindolones at the 6- and 7-positions in order to study this region of the active site and optimize noncovalent interactions. The electron density of derivative 28 bound to chicken PARP-1 revealed that the oxime makes a tight hydrogen bond with the catalytic gamma-carboxylate of glutamic acid (Glu) 988 in accordance with our original designs and models. Most of the compounds have been evaluated for inhibition of human PARP-1. Selected inhibitors were also tested for the ability to potentiate the cytotoxic effect of the DNA-damaging agent Topotecan.

Anti-oxidant activity of Centella asiatica on lymphoma-bearing mice.

Oral treatment with 50 mg X kg(-1) day(-1) of crude methanol extract of Centella asiatica for 14 days significantly increased the anti-oxidant enzymes, like superoxide dismutase (SOD), catalase and glutathione peroxidase (GSHPx), and anti-oxidants like glutathione (GSH) and ascorbic acid decreased in lymphoma-bearing mice.

Novel tricyclic poly(ADP-ribose) polymerase-1 inhibitors with potent anticancer chemopotentiating activity: design, synthesis, and X-ray cocrystal structure.

A series of novel compounds have been designed that are potent inhibitors of poly(ADP-ribose) polymerase-1 (PARP-1), and the activity and physical properties have been characterized. The new structural classes, 3,4,5,6-tetrahydro-1H-azepino[5,4,3-cd]indol-6-ones and 3,4-dihydropyrrolo[4,3,2-de]isoquinolin-5-(1H)-ones, have conformationally locked benzamide cores that specifically interact with the PARP-1 protein. The compounds have been evaluated with in vitro cellular assays that measure the ability of the PARP-1 inhibitors to enhance the effect of cytotoxic agents against cancer cell lines.

Recognition of follicle stimulating hormone (alpha-subunit) by a recombinant receptor protein domain coded by an alternately spliced mRNA and expressed in Escherichia coli.

To assess the functional significance of putative proteins encoded by alternately spliced mRNA of the sheep testicular FSH receptor, a short form cDNA comprising of the first four exons (117 residues mature protein) was engineered for expression in Escherichia coli. The expressed protein of molecular mass 15 kDa was purified to homogeneity and verified by reaction with an antibody against a synthetic peptide sequence unique to the amino (N)-terminal region FSH receptor. The purified FSH receptor domain protein bound 125I-labeled hFSH in a ligand blot on polyvinylidine difluoride membranes. Further analyses by slot blot revealed high affinity of the immobilized protein with significant reaction at 10 pmol. As the immobilized receptor protein also reacted with structurally related hormones (125I-labeled LH/125I-labeled human chorionic gonadotropin), we confirmed that interaction most probably occurred via the common alpha-subunit of these glycoprotein hormones. Our results reveal that this N-terminal portion of the FSH receptor contain(s) major site(s) for hormone recognition that could be mediated via the alpha-subunit. A rabbit antibody to the receptor inhibited FSH action in receptor bearing cells, revealing the utility of such recombinant FSH receptor protein(s) for modulation of hormone action.

Metabolism of proteins and glycoproteins in tumour bearing mice treated with Aeromonas L-asparaginase.

L-asparaginase, isolated in our laboratory, from Aeromonas had been found to be antileukaemic. In the present study, changes in the levels of proteins and glycoproteins in leukaemic mice and under treatment with Aeromonas L-asparaginase have been compared. Levels of protein bound hexose, fucose and sialic acid which were increased during leukaemia attained normal levels when treated with L-asparaginase. The increased blood urea level declined significantly during enzyme therapy. Effects of L-asparaginase are compared with 'Leunase', a commercially available drug used in the treatment of leukaemia.

Acute renal failure in asphyxiated newborns.

The study was undertaken to evaluate the occurrence of renal failure following perinatal asphyxia in the newborns. Thirty newborns with severe birth asphyxia were included in the study along with 30 normal newborns who comprised the control group. Any neonate presenting with oliguria or blood urea more than 40 mg/dl or creatinine more than 1 mg/dl was subjected to a fluid and diuretic challenge. If oliguria or renal dysfunction persisted then the child was labelled as renal failure and these subjects were further investigated. It was observed that 43% of asphyxiated babies developed acute renal failure (ARF); 69.2% babies had oliguric renal failure. While no significant correlation could be seen between Apgar scores at 5 and 10 min and development of ARF, a significant relationship was seen between hypoxic-ischemic encephalopathy and ARF. Patients with oliguric ARF carried a poorer prognosis as compared to non-oliguric ARF.

Renal dysfunction in septicemic newborns.

The study was undertaken to assess the association and incidence of acute renal failure (ARF) in septicemic neonates. Thirty neonates with septicemia formed the subject matter. Neonates with renal dysfunction were labelled as ARF patients after non responsiveness to a fluid and a diuretic challenge. Renal function tests were also evaluated. Nearly 15% neonates with septicemia developed ARF which was predominantly oliguric in type. The mortality rate in the septicemic neonates with ARF was significantly high. Further the mortality in neonates with oliguric ARF was significantly higher than those with non-oliguric ARF.