Pralhad Kushtagi

Preparation and, in vitro, preclinical and clinical studies of aceclofenac spherical agglomerates

Aceclofenac agglomerates were prepared by spherical crystallization technique using a three solvent system comprising acetone: dichloromethane (DCM): water (bridging liquid, good solvent and bad solvent, respectively). Hydroxypropyl methylcellulose-50cps (HPMC) in different concentrations was used as hydrophilic polymer. The effect of speed of rotation and amount of bridging liquid on spherical agglomeration were studied. The agglomerates were subjected to various physicochemical evaluations such as practical yield, drug content, particle size, loss on drying, porosity, IR spectroscopy, differential scanning calorimetry, X-ray diffraction studies, relative crystallinity, scanning electron microscopy, micromeritic properties, solubility and dissolution studies. The agglomerates showed improved micromeritic properties as well as dissolution behaviour in comparison to conventional drug crystals. The optimized agglomerates (F-9) showed good sphericity as well as high drug release, and hence they were compressed into tablets by direct compression. The tablets were found within the limits with respect to various physicochemical parameters. The dissolution rate of prepared tablets was better than that of marketed tablet and pure drug. The optimized agglomerates and tablet formulations were found to be stable for 6 months under accelerated conditions. The in vivo studies (preclinical pharmacokinetics, pharmacodynamics and toxicity studies, and clinical pharmacokinetics) of optimized agglomerates were carried out. The results of preclinical studies revealed that the agglomerates provided improved pharmacodynamic and pharmacokinetic profiles of drug besides being nontoxic. The results of pharmacokinetic studies of optimized tablet in human subjects indicated improved pharmacokinetic parameters of drug in comparison with that of marketed tablet.

Simultaneous detection of periodontal pathogens in subgingival plaque and placenta of women with hypertension in pregnancy

BackgroundThere are many studies documenting increased prevalence of periodontal infection in women with preeclampsia. But, very few studies have attempted to establish causal relationship between the two.ObjectiveTo find out causal circumstantial evidence by isolating specific periodontal pathogens in oral and placental samples.Materials and methodsAntenatal periodontal screening and subgingival plaque collection was carried out in ten women with hypertension in pregnancy and ten normotensive controls on their hospital admission at term for cesarean delivery. Placental biopsy was obtained after aseptic placental collection at the time of elective cesarean delivery. Subgingival plaque and placental biopsy were studied for Porphyromonas gingivalis, Fusobacterium nucleatum, Treponema denticola, Prevotella intermedia and Aggregatibacter actinomycetemcomitans using quantitative polymerase chain reaction technique.Periodontist and laboratory personnel were unaware of case or control status. Periodontal status was not informed to the obstetrician recruiting the cases and laboratory. Microbiology report was not revealed till end of the study.ResultsPeriodontal pathogens were found to be high in the group with hypertension than the controls. P gingivalis was found in all the samples from subgingival plaque and placenta, irrespective of the periodontal disease status.ConclusionIn cases with hypertension, periodontal pathogens are present in higher proportion in subgingival plaque and placenta.

DNA Promoter Methylation-dependent Transcription of the Double C2-like Domain β (DOC2B) Gene Regulates Tumor Growth in Human Cervical Cancer

Background: DOC2B promoter hypermethylation is an early and frequent event in cervical cancer. Results: DOC2B hypermethylation induces transcriptional repression, reactivated by demethylation; ectopic expression increases Ca2+ flux and inhibits key characteristics of tumorigenesis including proliferation, motility, and invasion. Conclusion: DOC2B gene is epigenetically regulated and inhibits cervical cancer growth. Significance: DNA methylation regulates DOC2B gene expression in cervical cancer. Double C2-like domain β (DOC2B) gene encodes for a calcium-binding protein, which is involved in neurotransmitter release, sorting, and exocytosis. We have identified the promoter region of the DOC2B gene as hypermethylated in pre-malignant, malignant cervical tissues, and cervical cancer cell lines by methylation-sensitive dimethyl sulfoxide-polymerase chain reaction and bisulfite genome sequencing; whereas, it was unmethylated in normal cervical tissues (p < 0.05). The promoter hypermethylation was inversely associated with mRNA expression in SiHa, CaSki, and HeLa cells and treatment with demethylating agent 5-aza-2-deoxycytidine restored DOC2B expression. The region −630 to +25 bp of the DOC2B gene showed robust promoter activity by a luciferase reporter assay and was inhibited by in vitro artificial methylation with Sss1 methylase prior to transient transfections. Overexpression of the DOC2B gene in SiHa cells when compared with controls showed significantly reduced colony formation, cell proliferation, induced cell cycle arrest, and repressed cell migration and invasion (p < 0.05). Ectopic expression of DOC2B resulted in anoikis-mediated cell death and repressed tumor growth in a nude mice xenograft model (p < 0.05). DOC2B expressing cells showed a significant increase in intracellular calcium level (p < 0.05), impaired AKT1 and ERK1/2 signaling, and induced actin cytoskeleton remodeling. Our results show that promoter hypermethylation and silencing of the DOC2B gene is an early and frequent event during cervical carcinogenesis and whose reduced expression due to DNA promoter methylation may lead to selective cervical tumor growth.

Effectiveness of subcutaneous injection of sterile water to the lower back for pain relief in labor

The analgesic efficacy of subcutaneous injection of sterile water compared to isotonic saline was investigated in a randomized, controlled study on a total of 100 women in the active phase of labor and who complained of low back pain. Pain perception was rated on a numerical rating scale before and at 10 and 45 minutes after the injection. The initial pain score was the same in both groups and pain relief was expressed by both groups irrespective of the solution injected, but the sterile water group had significantly higher relief scores compared to those receiving saline. This was not influenced by maternal age, parity, education, body mass index, cervical dilatation at intervention or fetal size, suggesting that subcutaneous injection of sterile water to the lower back provides relief from back pain during labor.

Maternal mid-pregnancy serum triglyceride levels and neonatal birth weight

Although disturbances in maternal glucose metabolism and resultant fetal hyperinsulinemia are known to significantly impact fetal overgrowth, it has been suggested that other fuels such as lipids and amino acids may be the determinants of fetal growth in nondiabetics. Maternal serum lipid levels increase in mid to late pregnancy; and in women with gestational diabetes, elevated triglycerides were significantly associated with birth weight corrected for gestational age. Although the role of maternal hyperlipidemia in fetal growth is not known, maternal fasting serum triglyceride levels at 24 to 32 weeks of Pregnancy were shown to be significantly and positively associated with neonatal weight at term,independent of maternal prepregnancy obesity, weight gain during pregnancy, or mid-pregnancy plasma glucose levels (either fasting or postprandial) in Japanese and white women. The aim of the present study was to determine the relationship between maternal mid-pregnancy serum lipid levels and neonatal birth weight in our region. Fasting levels of serum triglycerides and total cholesterol were evaluated in 108 consenting consecutive women who attended for prenatal care with a singleton pregnancy between 24 to 34 weeks and who had abnormal results on the glucose challenge test (plasma glucoseN140 mg/dL) and normal results from the oral 100 g 3-hour glucose tolerance test. Women with antiphospholipid antibody syndrome, hypertension, or thyroid disorder were excluded. The relationship between mid-pregnancy triglyceride and total cholesterol values and neonatal birth weight was determined according to fasting glucose, body mass index (BMI, calculated as weight in kilograms divided by height in meters squared) and pregnancy weight gain—with 95 mg/dL, 25, and 10 kg as the cut-off values, respectively. The relative risk of having heavier babiesweighing 3.5 kg or greater was 2 (95% CI, 0.9–4.2) for women with hypertriglyceridemia and 2.6 (95% CI, 0.78–8.4) when controlled for fasting glucose, BMI, and pregnancy weight gain. When studied together, the combination of hypertriglyceridemia, fasting hyperglycemia, and higher BMI was a significant predictor of heavier neonates. It is important to note that the higher relative risk for heavier babies in women with hypertriglyceridemia persisted even when the variables such as greater BMI, greater pregnancy weight gain, and hyperglycemia were accounted for. With a combination of greater BMI and fasting hyperglycemia, women with hypertriglyceridemia had a much higher relative risk for heavier babies (RR 9.7; 95% CI, 1.7–62.6).

Periodontal infection in women with low birth weight neonates

It has been hypothesized that in the presence of periodontal disease, periodontal pathogens are disseminated hematogenously to target the placenta,membranes, and fetus. This bacterial challenge may result in increased cytokine expression, and affect placental function and precipitate preterm labor. The present case-control study was designed to investigate whether women who delivered low birth weight neonates had a higher rate of periodontal infection. Material and Methods: A total of 150 consecutive women admitted in labor (preterm or with evidence of fetal growth restriction) with known gestational age and a first trimester ultrasound dating The control group consisted of women who had delivered a neonate of appropriate size for gestational age at term (≥37–41 weeks), matched to each case participant for age, parity, and socioeconomic status. Oral examinations were carried out within 72 hours of delivery by a periodontist blinded to the case-control status. Results: Periodontal infection was noted in 59% (177/300) of women in the study. The presence of periodontal infection was significantly higher in women with preterm (92.1%; 82/ 89 cases) and growth restricted neonates (91.8%; 56/61 cases) compared with women in the control group (26%; 39/ 150 cases) (p<0.001). All clinical parameters considered to indicate periodontal infection showed a significant association with low birth weight neonates. These parameters, whether used individually or inferred together, were indicative of periodontal infection and correlated with pregnancy outcomes. Conclusion: The findings that periodontal infection is prevalent in pregnancy and significantly higher in women who delivered low birth weight neonates suggest a need for developing preventive strategies. The clinical diagnosis of periodontal infection correlated with either inflammatory mediators or microbiological confirmation would add validity to the claims of a causal relationship.

Mitochondrial DNA Variation Analysis in Cervical Cancer

This study was undertaken to investigate the mitochondrial DNA (mtDNA) variation in non-malignant and malignant cervical tissue samples. We have identified 229 and 739 variations non-malignant and malignant tissues respectively distributed over 321 locations in the D-loop (50 in non-malignant and 166 in malignant; 216 variations), coding region (139 in non-malignant and 455 in malignant; 594 variations) tRNA and rRNA genes (39 in non-malignant and 119 in malignant; 158 variations). Besides, 77 novel and 34 various other disease associated variations were identified in non-malignant and malignant samples. A total of 236 tumor specific variations in 201 locations representing 30.1% in D-loop, 59.3% in coding regions and 10.6% in RNA genes were also identified. Our study shows that D loop (in 67 locations) is highly altered followed by ND5 (35 locations) region. Moreover, mtDNA alterations were significantly higher in malignant samples by two tailed Fishers exact test (P≤0.05) with decreased mtDNA copy numbers. Bioinformatic analysis of 59 non-synonymous changes predicted several variations as damaging leading to decreased stability of the proteins. Taken together, mtDNA is highly altered in cervical cancer and functional studies are needed to be investigated to understand the consequence of these variations in cervical carcinogenesis and their potential application as biomarkers.

Periodontal Disease and Pregnancy Hypertension: A Clinical Correlation

BACKGROUND Periodontal disease is thought to be associated with increased risk of systemic diseases and adverse pregnancy outcomes, including pregnancy hypertension (PH). The aim of the present study is to find the prevalence of periodontal disease in females with PH in a rural-based medical institute. METHODS The present case control study was performed with 200 females, 100 with PH and 100 without PH. Antenatal periodontal screening was performed within 72 hours of their hospital admission for delivery. The periodontal parameters assessed were oral hygiene index-simplified, gingival index, mean probing depth, and loss of attachment. RESULTS Prevalence of periodontal disease was 65.5% and was significantly higher (P <0.0001) in females with hypertension (relative risk = 1.5; 95% confidence interval [CI] = 1.3 to 1.9). Socioeconomic status, education, occupation, and body mass index did not appear to influence the relationship between periodontal disease and PH on bivariate multiple logistic regression analysis. Nulliparous females were at higher odds to develop periodontal disease and PH (odds ratio = 1.7; 95% CI = 0.5 to 6.1). As the severity of periodontal disease increased from moderate to severe, the severity of hypertension also increased (r(2) = 0.8 and 0.5 for moderate and severe periodontal disease, respectively). CONCLUSION Periodontal disease is more prevalent in females with PH.

Protein profile study of clinical samples of ovarian cancer using High Performance Liquid Chromatography-Laser Induced Fluorescence (HPLC-LIF)

New techniques for the early detection of cancer are fast emerging. This is essential for more effective diagnosis and control of the disease. We have used a High Performance Liquid Chromatography-Laser Induced Fluorescence (HPLCLIF) technique to record chromatograms of proteins in serum and ovarian tissue samples. The recorded chromatograms of normal, benign and malignant samples were analyzed using statistical (Principal Component Analysis) methods. It is shown that chromatograms of the samples can be classified into sets, and a model based on such a classification can be used to analyze protein profiles of test samples of serum and ovarian tissue for the detection of malignancies.

Evaluation of two uterotonic medications for the management of the third stage of labor.

It is perhaps surprising that as yet no consensus exists among clinicians concerning the best way to prevent postpartum hemorrhage. The present study was conducted as an attempt to evaluate the scope of 125 µg (note the dosage) of carboprost tromethamine given intramuscularly in comparison with intravenous methyl ergometrine. Two hundred and fifteen parturients were randomly assigned to use either drug at the time of delivery of anterior shoulder of the baby. The outcome measures were duration of third stage the measured blood loss after placental delivery and the incidence of side effects. Amount of blood loss was quantified by noting the increment in weight of standardised tampons which were placed high up in the vagina immediately after placental delivery. Weight of blood in grams was considered equal to volume in ml. (excerpt)