Design, synthesis and molecular docking of pyrazolo [3,4d] thiazole hybrids as potential anti-HIV-1 NNRT inhibitors.

A series of pyrazolo[3.4,d]thiazole hybrids 6 were synthesized from 5-arylidene-2-imino-3-(4-arylthiazol-2-yl)-thiazolidin-4-ones 5. The 5-arylidene-2-imino-3-(4-arylthiazol-2-yl)-thiazolidin-4-ones 5 were synthesized from 2-amino-4-arylthiazoles 1 and 2-chloro-acetamido-4-arylthiazoles 2 via the formation of 2-imino-3-(4-substituted-arylthiazol-2-yl)-thiazolidin-4-ones 3 using substituted aldehydes 4. The 5-acrylidene derivative 5 on cyclisation with phenyl hydrazine give the pyrazolo [3, 4, d] thiazole derivatives 6. The obtained pyrazolo [3.4, d]thiazole derivatives were studied as anti-HIV-1 NNRT inhibitors. It was found that these compounds might have potent RT inhibition activity.

Autophagy and oral cancers: A short review.

Autophagy is a catabolic process, which involves recycling of cellular elements and it seems to play a major role in etiopathogenesis of cancers. Autophagy is thought to buffer metabolic stress, thereby aiding in cell survival. Also, it is found that inhibiting autophagy under deficient nutrition can restore cell death to apoptosis. Therefore, autophagy plays a dual role in cancer therapeutics. Globally oral cancer is a major concern of numerous deaths, so comprehensive work needs to be undertaken in the field of its progression and to determine positive treatment modalities. There exists a serious dearth of knowledge to understand and comprehend the process of autophagy, and it is beyond doubt that further research in this field would bring forth several new methods of cancer prevention and treatment.

Pectoral nerve block1 versus modified pectoral nerve block2 for postoperative pain relief in patients undergoing modified radical mastectomy: a randomized clinical trial.

BACKGROUND: Pectoral nerve block1 (PEC1) given between pectoralis major and minor, and modified pectoral nerve block2 (mPEC2) performed between pectoralis minor and serratus anterior, can provide continuous analgesia after modified radical mastectomy (MRM) when catheters are placed before skin closure. This study was designed to compare PEC1 and mPEC2 block for providing postoperative pain relief after MRM. METHODS: Sixty-two physically fit patients undergoing MRM were assigned into two groups (Group PEC1, n=31 and Group mPEC2, n=31). Before wound closure, epidural catheter was placed in the group designated muscle plane and 30ml of 0.25% bupivacaine was injected through the catheter after wound closure. Bupivacaine 15ml of 0.25% top up was given on patient's demand or whenever visual analogue scale (VAS) score was>4. Time for first analgesia (TFA), number of top ups and VAS was recorded at 0.5, 6, 12, 18, 24 h after surgery. Sensory blockade was assessed 30 min after extubation. RESULTS: Analgesia was significantly prolonged in group mPEC2 [mean(SD)] 313.45(43.05) vs 258.87(34.71) min in group PEC1, P<0.001. Total pain experienced over 24 h was significantly less in group mPEC2 [mean(SD)] 9.77(6.93) than in group PEC1 24.19(10.81), P<0.0001. Consequently, top up requirements were significantly reduced in group mPEC2 than in group PEC1 [median(range)] 3(2-4) vs 4(3-5) respectively, P<0.001. Lateral pectoral (77.42% and 35.48%) and thoracodorsal nerves (93.55% and 48.39%) had higher incidence of sensory block in group mPEC2 than group PEC1, P<0.001. CONCLUSIONS: mPEC2 provides better postoperative analgesia than PEC1 when catheters are placed under direct vision after MRM. CLINICAL TRIAL REGISTRATION: CTRI/2017/02/007811 (REF/2015/11/010185).

Metabolomic and Genome-wide Association Studies Reveal Potential Endogenous Biomarkers for OATP1B1.

Transporter-mediated drug-drug interactions (DDIs) are a major cause of drug toxicities. Using published genome-wide association studies (GWAS) of the human metabolome, we identified 20 metabolites associated with genetic variants in organic anion transporter, OATP1B1 (P < 5 × 10-8 ). Of these, 12 metabolites were significantly higher in plasma samples from volunteers dosed with the OATP1B1 inhibitor, cyclosporine (CSA) vs. placebo (q-value < 0.2). Conjugated bile acids and fatty acid dicarboxylates were among the metabolites discovered using both GWAS and CSA administration. In vitro studies confirmed tetradecanedioate (TDA) and hexadecanedioate (HDA) were novel substrates of OATP1B1 as well as OAT1 and OAT3. This study highlights the use of multiple datasets for the discovery of endogenous metabolites that represent potential in vivo biomarkers for transporter-mediated DDIs. Future studies are needed to determine whether these metabolites can serve as qualified biomarkers for organic anion transporters. Quantitative relationships between metabolite levels and modulation of transporters should be established.

A Longitudinal HbA1c Model Elucidates Genes Linked to Disease Progression on Metformin.

One-third of type-2 diabetic patients respond poorly to metformin. Despite extensive research, the impact of genetic and nongenetic factors on long-term outcome is unknown. In this study we combine nonlinear mixed effect modeling with computational genetic methodologies to identify predictors of long-term response. In all, 1,056 patients contributed their genetic, demographic, and long-term HbA1c data. The top nine variants (of 12,000 variants in 267 candidate genes) accounted for approximately one-third of the variability in the disease progression parameter. Average serum creatinine level, age, and weight were determinants of symptomatic response; however, explaining negligible variability. Two single nucleotide polymorphisms (SNPs) in CSMD1 gene (rs2617102, rs2954625) and one SNP in a pharmacologically relevant SLC22A2 gene (rs316009) influenced disease progression, with minor alleles leading to less and more favorable outcomes, respectively. Overall, our study highlights the influence of genetic factors on long-term HbA1c response and provides a computational model, which when validated, may be used to individualize treatment.

Melatonin improves ultraviolet B-induced oxidative damage and inflammatory conditions in cutaneous tissue of a diurnal Indian palm squirrel Funambulus pennanti.

BACKGROUND: Skin is exposed to various abiotic and biotic factors. Solar radiation, of which ultraviolet (UV) rays are a principle component, increases the free radical load, and the accumulation of reactive oxygen species (ROS) causes lipid peroxidation, DNA damage and apoptosis, and is also associated with inflammatory responses recruiting molecules [nuclear factor (NF)-κB, interleukin (IL)-6] that can potentially further aggravate the damaged milieu of the cells. One of the potent causes of skin cancers is exposure to UV rays. UV radiation generates a wide range of biological responses such as adaptive, inflammatory and immunological reactions in the skin. OBJECTIVE: To examine the effects of pretreatment with melatonin on UVB (290-320 nm) radiation-mediated damage to the skin of a diurnal rodent Funambulus pennanti. RESULTS: The UVB radiation (1·5 J cm(-2) for 30 min daily on the shaved abdominal area) for 4 days caused a significant increase in the lipid peroxidation products (thiobarbituric acid reactive substances, TBARS) and decreased the activity of the antioxidant enzymes (superoxide dismutase, catalase and glutathione peroxidase) of the skin. Pretreatment with melatonin (100 µg 100 g(-1) bodyweight subcutaneously) improved the damage induced by UVB radiation on the skin and might act via a receptor-independent mechanism. No significant effect of melatonin pretreatment was found on the expression pattern of MT1 (melatonin membrane receptor) and RORα (nuclear retinoic orphan receptor alpha), which suggests a receptor-independent action. However, NF-κB and inflammatory cytokine IL-6 levels were downregulated in the squirrels pretreated with melatonin before the UVB radiation. CONCLUSION: UVB radiation induced oxidative stress in the skin culminating in an inflammatory response. The action of melatonin in protecting the skin from oxidative damage occurs in a receptor-independent manner by lowering the oxidative damage and inflammatory response. On the other hand, melatonin decreased the expression of NF-κB and the circulating proinflammatory cytokine IL-6, suggesting an anti-inflammatory action in protecting the skin from UVB radiation.

Primary aspergillosis of the larynx: a case report.

Aspergillosis of larynx is very rare. Only a few cases have been reported in medical literature. Larynx is generally involved secondarily by aspergillosis. Only rarely the larynx happens to be the primary site of involvement. The lesions may be confined to the vocal folds or may involve various other sites of the larynx. There is, invariably, certain risk factors which may predispose for fungal infection like immune deficiency condition, use of inhaled or systemic steroids, long and continuous use of antibiotics etc. These lesions may mimic malignancy or a premalignant condition. Awareness of this entity is essential as the management depends on accurate diagnosis. These lesions invariably respond well to antifungal therapy with elimination of risk factors. Here we have reported a case of primary aspergillosis of the larynx without involvement of other part of the airway and without any generalized immune deficit in a 35 year old woman.

Recurrent pregnancy loss in polycystic ovary syndrome: role of hyperhomocysteinemia and insulin resistance.

Recurrent pregnancy loss (RPL) in polycystic ovary syndrome (PCOS), which occurs in ∼50% of total pregnancies is a frequent obstetric complication. Among the several hypotheses, insulin resistance (IR), obesity and hyperhomocysteinemia (HHcy) play significant role/s in RPL. This study was conducted to assess the link between elevated levels of homocysteine and IR in PCOS-associated women with RPL in Kolkata, India. A retrospective study was conducted of one hundred and twenty six PCOS women (<30 years) who experienced two or more spontaneous abortions during the first trimester presenting to Institute of Reproductive Medicine (IRM) in Kolkata during the period of March 2008 through February 2011. One hundred and seventeen non-PCOS subjects with matching age range were randomly chosen as controls. Incidence of HHcy and IR was 70.63% (n = 89) and 56.34% (n = 71), respectively, in RPL-affected PCOS population which was significantly higher (p<0.04; p<0.0001) when compared to the non-PCOS set (HHcy: 57.26%; IR: 6.83%). Rates of miscarriage were significantly higher (p<0.008; p<0.03) in hyperhomocysteinemia-induced miscarriage when compared to the normohomocysteinemic segment (PCOS: 70.63% vs.29.36% & non-PCOS: 57.26% vs. 42.73%) along with the insulin resistant (p<0.04; p<0.0001) population (PCOS: 70.63% vs. 56.34% & non-PCOS: 57.26% vs. 6.83%) in both groups. A probabilistic causal model evaluated HHcy as the strongest plausible factor for diagnosis of RPL. A probability percentage of 43.32% in the cases of HHcy- mediated RPL suggests its increased tendency when compared to IR mediated miscarriage (37.29%), further supported by ROC-AUC (HHcy: 0.778vs. IR: 0.601) values. Greater susceptibility towards HHcy may increase the incidence for miscarriage in women in India and highlights the need to combat the condition in RPL control programs in the subcontinent.

Metaphase arrest and delay in cell cycle kinetics of root apical meristems and mouse bone marrow cells treated with leaf aqueous extract of Clerodendrum viscosum Vent.

OBJECTIVES: To study cell cycle delay and metaphase arresting activity of leaf aqueous extract of Clerodendrum viscosum Vent. (LAECV) in root apical meristems and mouse bone marrow cells. MATERIALS AND METHODS: Cell cycle delay and metaphase arresting activities of LAECV were analysed, in root apical meristems of onion and wheat, and in mouse bone marrow cells, by scoring mitotic index, metaphase frequency and transition of cells from metaphase to anaphase. Colchicine was used as the standard metaphase arresting drug. Phytochemicals present in LAECV were detected and their phytotoxic activity was evaluated by analysing green-gram (Vigna radiata) seedling's root growth retardation and branch root swelling phenomenon. RESULTS: LAECV treatment resulted in dose-dependent root growth retardation of green-gram seedling root length (P < 0.01) and half maximal growth inhibitory concentration (IC(50) ) of LAECV was 0.87 mg/ml at 144 h. In onion and wheat root meristem cells the mitotic index decreased, metaphase frequency increased and transition from metaphase to anaphase reduced. Experimentation with mouse bone marrow cells indicated that LAECV induced metaphase arrest (164.3% increase in arrested metaphases per 300 mg/kg body weight, over 2.5 h). Phytochemicals like carbohydrates, glycosides, saponins, terpenoids, triterpenoids, tannins and trace amounts of alkaloids were detected in LAECV. CONCLUSION: It may be said that LAECV contains mitostatic and metaphase arresting components that are able to induce significant metaphase arrest in root apical meristems and also in mouse bone marrow cells.

Altered trace mineral milieu might play an aetiological role in the pathogenesis of polycystic ovary syndrome.

Insulin resistance is a very common associate of polycystic ovary syndrome (PCOS). Pathophysiology in relation with the essential elements including copper, magnesium, zinc, manganese, chromium, and calcium has been reported in women with insulin resistance. This prospective study was designed to explore whether the women with PCOS do exhibit altered serum element levels in association with/without insulin resistance. One hundred and thirty-two women with PCOS and forty-six control women were studied. Women with PCOS were further divided based on the presence of insulin resistance (insulin resistant: n = 50; non-insulin resistant: n = 82). In all women, basal levels of gonadotropins, prolactin, testosterone, insulin, glucose, and the six different elements were measured. Serum levels of testosterone (p < 0.001), luteinizing hormone (p < 0.05), and fasting insulin (p < 0.004) were significantly higher in the PCOS population compared to controls as well as PCOS women without insulin resistance. Women with PCOS exhibited a significantly high calcium (p < 0.04) and lower manganese levels (p < 0.002) when compared to controls. However, the PCOS women with insulin resistance exhibited significantly lower serum levels of magnesium and chromium (p < 0.04), in addition to higher levels of zinc and copper (p < 0.04). The differences in calcium (p < 0.03) and manganese levels (p < 0.0001) became aggravated with the presence of insulin resistance when compared to control as well as PCOS women without insulin resistance. In PCOS-associated insulin resistance, circulating serum magnesium (r = -0.31; p < 0.03) and chromium (r = -0.38; p < 0.006) status significantly correlated with fasting insulin levels. We conclude that imbalanced element status may be a key foundation for insulin resistance in PCOS. The findings in this study should be investigated with further trials in order to obtain new insights into PCOS.

Bioactive constituents of Homalomena aromatica essential oil and its antifungal activity against dermatophytes and yeasts.

Homalomena aromatica rhizomes are rich source of essential oils, which have been attributed for various medicinal uses. In the present investigation, essential oil from H. aromatica rhizomes was isolated and subjected to gas chromatography-mass spectrum (GC-MS) analysis. Fifty-five chemical constituents were reported from H. aromatica rhizomes of which T-muurolol (5.32%), viridiflorol (3.69%), α-selinene (2.19%), M-cymene (2.19%) and γ-Muurolene (1.81%) were identified and reported for the first time. Other major components were identified as linalool (62.5%), terpene-4-ol (7.08%), δ-cadinene (5.57%), α-cadinol (3.71%) and spatulenol (1.81%). H. aromatica rhizome essential oil showed high antimicrobial activity against Trichophyton rubrum, Trichophyton mentagrophytes, Microsporum fulvum, Microsporum gypseum, Trichosporon beigelii and Candida albicans.

The potential risk factors and the identification of hearing loss in infants.

To identify potential risk factors for hearing loss from the infant with high risk criteria as guided by the Joint Committee on Infant Hearing (JCIH, 2000). 490 infants with age range of 2 days to 6 months who had undergone detailed audiological evaluation during for the period of 3 years at Gauhati Medical College Hospital were taken for the study. The hearing screening was performed on each infant using Distortion Product Otoacoustic Emission (DPOAE) testing. The infants who failed DPOAEs screening were followed up and Auditory Brainstem Response testing. Out of 490 high risk infants who had undergone detailed audiological evaluation 145 infants were found to be having hearing loss. Out of 145 infants 73 infants were male and 72 infants were female. The risk factor for hearing loss with the highest incidence was hyperbilirubenemia, Apgar scores of 0-4 at 1 min or 0-6 at 5 min was the second most prevalent risk factor, followed by TORCH infections. This study suggests the need for review of high risk register that is used along with the physiological and electrophysiological hearing test to screen the infants. The high risk register remains helpful in determining follow up plans so that children who may develop late onset of hearing loss will not be missed. Being aware of which risk factors are more likely to cause hearing loss in infants would be helpful to plan for follow up these children.

A novel method of plasmid isolation using laundry detergent.

Since the discovery of plasmid, various methods have been developed to isolate plasmid DNA. All the methods have one common and important target of isolating plasmid DNA of high quality and quantity in less time. These methods are not completely safe because of use of toxic chemicals compounds. The developed protocol for plasmid extraction is based on the alkaline lysis method of plasmid preparation (extraction atpH 8.0) with slight modifications. Cell lysis reagent sodium dodecyl sulfate is replaced by lipase enzyme present in laundry detergent. A good plasmid preparation can be made, which is well suited for subsequent molecular biology applications. By taking safety measures on count, contaminants like, RNA and protein can be completely avoided with maximized plasmid yield. The resultant plasmid quality and quantity can be well comparable to other prevalent methods.

Clinical outcome of levonorgestrel intra-uterine system in idiopathic menorrhagia.

AIMS: To evaluate the efficacy, acceptability and side effects of levonorgestrel intrauterine system in patients with idiopathic menorrhagia. METHODS: A prospective, non-randomized clinical study in which 42 women between the age of 35-55 years complaining of menorrhagia with or without irregular cycle were recruited during one year period. The women were followed up at 3, 6, 12, 24 and 36 months after insertion. RESULTS: The reduction in mean blood loss was found to be statistically significant with a p value of < 0.001 for all the follow-up periods. CONCLUSION: The present study projects levonorgestrel intrauterine system as an effective patient friendly device with a high degree of compliance and worth considering as a viable alternative to surgery for the management of menorrhagia due to dysfunctional uterine bleeding in developing countries like India.

Evaluation of colposcopy vs cytology as secondary test to triage women found positive on visual inspection test.

BACKGROUND: Colposcopic evaluation and guided biopsy is an important diagnostic step and standard of management for abnormal cytology smears in developed countries. AIM: The present study evaluates the performance of colposcopy vs conventional cytology in estimating the presence and grade of cervical disease against the reference standard of histopathology as a secondary test modality to triage women found positive on primary screening by visual inspection with 5% acetic acid (VIA). SETTINGS AND DESIGN: Colposcopy and directed biopsy were performed after primary screening for cervical cancer in an urban hospital cancer screening clinic between January 2000 and June 2005, in Mumbai, India. MATERIALS AND METHODS: Healthy women (8863) in the age group of 35-65 years participated in the cervical cancer early detection program in the hospital and community cancer screening clinics in Mumbai. Women found positive on the primary screening test by VIA underwent diagnostic evaluation by Pap smear cytology and colposcopy evaluation with directed biopsies. STATISTICAL ANALYSIS: Accuracy parameters and their 95% confidence intervals were calculated using 2 X 2 tables and standard formulae. RESULTS: The test range of sensitivity of colposcopy for the detection of histologically confirmed cervical intraepithelial neoplasia grade 1+ (CIN1+) or CIN2+ was 58.0-74.7% and that of specificity was 57.5-92.9%. The sensitivity and specificity of cytology to detect CIN2+ was 57.4% and 99.4%, respectively. CONCLUSION: Colposcopy is a good sensitive test for the detection of CIN and can be considered as a secondary testing tool to triage women found positive on VIA.

Relative potencies of natural estrogens on vitellogenin and choriogenin levels in the Indian freshwater spotted snakehead, Channa punctata: in vivo and in vitro studies.

The relative efficacies of three natural estrogens viz., estrone (E(1)), estradiol-17beta (E(2)) and estriol (E(3)) to induce synthesis of vitellogenin (Vg) and choriogenin (Chg) were assessed in primary hepatocyte cultures of the Indian freshwater spotted snakehead, Channa punctata. Hepatocytes were isolated from the spotted snakehead liver by a non-enzymatic protocol. Optimum culture conditions were standardized for ensuring their viability and functioning. Isolated hepatocytes were cultured for 48 h for monolayer formation and then exposed to various concentrations (0.001-10 microM) of the three estrogens. Competitive homologous ELISAs, developed and validated for spotted snakehead Vg and Chg were employed to determine the amounts of these two proteins secreted into the culture medium after 48 h of incubation. The results reveal that although all the three estrogens were effective in inducing the production of Vg and Chg in a dose-dependent manner, there were differences in their relative potencies. Of three estrogens, E(1) was the least potent and could induce synthesis of Vg and Chg only at a minimum concentration of 0.5 microM; whereas significant levels of both the proteins were quantified in culture medium by exposing the hepatocytes to E(2) or E(3) even at a concentration of 0.001 microM. All three estrogens were effective in inducing synthesis of Vg and Chg in vivo also. These results suggest the possibility of employing the above in vitro experimental design to monitor the presence of estrogens/estrogen-like chemicals in natural waters, which could interfere with the estrogen receptor system of fish. This study further points to the possibility of using Chg, in addition to Vg, as a parameter for screening various chemicals for their estrogenic activity.

Optimizing results of breast cancer surgery: a 8 yrs' experience.

Management of breast cancer in developing country, in contrast to Western countries, is challenging considering more number of advanced cases and poor understanding of the disease and its management by the population and even many physicians. We tried to look prospectively at our experience with breast cancers, trying to optimize the results given the wide- variety of cases we see.

Rare site giant cell tumors: report of two cases on phalanges of the finger and review of literature.

Giant cell tumor (GCT) of bone arising from a phalanx of a finger is extremely rare. We report two cases of GCT arising from a phalanx of a finger. One case presented with recurrence following the amputation of the left ring finger (performed elsewhere). He was treated successfully with ray amputation. The other case was treated primarily by intralesional curettage and autogenous bone graft. At their most recent follow-ups (80 and 24 months, respectively), both were recurrence free and had returned to their previous occupational and recreational activities.

Establishment of ELISA for murrel vitellogenin and choriogenin, as biomarkers of potential endocrine disruption.

Vitellogenin (Vg) and choriogenin (Chg) are sensitive biomarkers for testing endocrine disruption in fish. Therefore, we have developed immunoassays for Vg and Chg in the Indian freshwater murrel, Channa punctatus. Vg is a known precursor of egg-yolk proteins, whereas Chg contributes to the formation of egg-envelope. Vg and Chg were induced in male murrel by administration of estradiol-17beta. Chg had an apparent native molecular mass of 180 kDa. It consisted of a single peptide with a molecular mass of 110 kDa, whereas native Vg protein (530 kDa) contained 175 kDa peptide. Highly specific polyclonal antibodies against purified plasma proteins, Vg and Chg, were employed for developing competitive enzyme linked immunosorbent assays (ELISAs). The sensitivity of Vg assay was 3.9 ng/mL (working range 15-500 ng/mL) and of Chg assay was 1.56 ng/mL (working range 6-200 ng/mL). The inter- and intra-assay variations were well within acceptable limits. The two antisera did not cross-react with male plasma proteins. Antiserum to Vg did not cross-react with Chg. Similarly, antiserum to Chg showed no correlation with Vg. Further, immunofluorescence and Western blotting confirmed the specificity of Vg and Chg antisera.

Isolation and characterization of embryonic stem cell-like cells from in vitro-produced buffalo (Bubalus bubalis) embryos.

This study was carried out to isolate and characterize buffalo embryonic stem (ES) cell-like cells from in vitro-produced embryos. Inner cell mass (ICM) cells were isolated either mechanically or by enzymatic digestion from 120 blastocysts whereas 28 morulae were used for the isolation of blastomeres mechanically. The ICM cells/ blastomeres were cultured on mitomycin-C-treated feeder layer. Primary cell colony formation was higher (P < 0.05) for hatched blastocysts (73.1%, 30/41) than that for early/expanded blastocysts (25.3%, 20/79). However, no primary cell colonies were formed when blastomeres obtained from morulae were cultured. Primary colonies were formed in 14.1% (12/85) of intact blastocyst culture, which was significantly lower (P < 0.05) than that of 41.6% for ICM culture. These colonies were separated by enzymatic or mechanical disaggregation. Using mechanical disaggregation method, the cells remained undifferentiated and two buffalo ES cell-like cell lines (bES1, bES2) continued to grow in culture up to eight passages. However, disassociation through enzymatic method resulted in differentiation. Undifferentiated cells exhibited stem cell morphological features, normal chromosomal morphology, and expressed specific markers such as alkaline phosphatase (AP) and Oct-4. Cells formed embryoid bodies (EBs) in suspension culture; extended culture of EBs resulted in formation of cystic EBs. Following prolonged in vitro culture, these cells differentiated into several types of cells including neuron-like and epithelium-like cells. Furthermore, the vitrified-thawed ES cell-like cells also exhibited typical stem cell characteristics. In conclusion, buffalo ES cell-like cells could be isolated from in vitro-produced blastocysts and maintained in vitro for prolonged periods of time.