Anti-Histone H3.3 G34W antibody is a sensitive and highly specific immunohistochemistry marker for the diagnosis of Giant cell tumor of bone. A validation based on analysis of 198 cases from a single centre in India

Context: The diagnosis of giant cell tumor of bone (GCTB) is difficult in small biopsies with unusual age of presentation, location, and extensive secondary changes. Most of the GCTBs harbor H3F3A G34W mutations with a subset of cases showing alternate G34V, G34R, and G34L mutations. Objectives: To analyze the expression of anti-histone H3.3G34W antibody in different cellular components of GCTB across different locations and presentations (including the unusual ones) and validate the utility of this antibody in the diagnosis of GCTB and differentiate it from the other osteoclast-like giant-cell-rich lesions. Design: Immunohistochemistry was performed using anti-histone H3.3G34W antibody in the diagnosed cases of GCTB (136 cases of GCTB from 133 patients, including two malignant GCTBs) and other giant cell-containing lesions (62 cases). The presence of unequivocal crisp nuclear staining was considered positive. Results: Immunohistochemistry revealed unequivocal nuclear positivity in the mononuclear cells in 87.3% of the cases of GCTB. Of these, most showed diffuse expression with moderate to strong intensity staining. The positive staining was restricted to the nuclei of mononuclear cells with the nuclei of osteoclastic giant cells being distinctly negative. In addition to conventional GCTBs, two cases each of multicentric and malignant GCTB showed positive staining. The other giant-cell containing lesions were distinctly negative. The present study showed a sensitivity of 87.3% with specificity and positive predictive value of 100%. Conclusion: The anti-histone G34W antibody is a highly sensitive and specific marker for the diagnosis of GCTB and differentiating it from its mimics. The positive staining is restricted to the mononuclear cell component of GCTB with sparing the osteoclastic giant cells further reiterating the fact that the mononuclear stromal cells are the true neoplastic component of GCTB.

Right Sided Haemorrhagic Pleural Effusion Secondary to Pancreatitis

A 28-year-old male patient, painter by occupation came to the department of pulmonology of Rajarajeswari Medical College, Kambipura, Karnataka, with complaints of right sided chest pain since 4 months, dry cough since one month and loss of appetite with significant weight loss. The patient was a chronic alcoholic and a non-smoker. History revealed that he was admitted in a primary health care centre for one week with similar complaints 3 months back and chest radiographs were taken. The radiograph showed right sided massive pleural effusion for which diagnostic thoracocentesis was done, which revealed haemorrhagic fluid. Pleural fluid analysis showed lymphocytic predominance; there after, repeated thoracocentesis was done in the same primary health centre and up to 5 litres of haemorrhagic pleural fluid was aspirated. Routine blood reports were within normal limits.

Level of physical activity, determinants and barriers to motivate physical activities among south Indian diabetic patients: a prospective, questionnaire based study

Background: Several clinical trials have been reported that sedentary lifestyle modification including physical activity (PA) and weight loss significantly alters the sort-term as well as long-term incidence of diabetes. The present study explored the level of physical activity and its barriers among south Indian naïve patients who are attending the tertiary care teaching hospital.Methods: A prospective observational, questionnaire-based study. Patients with diabetes (both Type-1 and 2) who were attended diabetic clinic in the Department of General Medicine, Maharaja Institute of Medical Sciences (MIMS), Vijayanagaram, Andhra Pradesh, India was selected and recruited. Demographic data, clinical data were collected from the study participants. International Physical Activity questionnaire was used to determine each patient level of physical activity (PA). Physical activity is graded into low (<600 metabolic equivalent (MET)), moderate (600-3000 MET) and high (>3000 MET).Results: A total of 300 subjects were completed the entire study. Majority (62.7%) of the patients were suffering from Type-2 diabetes mellitus. Both male and female study subjects had low physical activity (70.5% and 74.1%) and moderate PA was seen in only one thirds of the patients. Patients with low physical activity had low glycaemic control compared to patients who were involved in moderate PA. Moderate PA patients had adequate glycaemic control (65.9 vs 34.1%, P <0.001). A total of 42.0% of subjects were suffering from one of the joint pains and friction joints, due to the following reason subjects were not involved in the PA.Conclusions: Low level of physical activity was observed in the study population, which is a risk factor for several micro-vascular complications over a period of time. It is very important to address the barriers of PA and vigorous counselling is needed which directs towards increasing PA.

Effect of statins treatment on glycemic status and development of new onset diabetes mellitus in a tertiary care teaching hospital: a cross sectional study

Background: Several observational studies, well controlled randomized trials and meta-analyses reported that patients treated with statins has high risk of new onset diabetes mellitus (NODM), but the exact incidence and mechanism is still unclear and controversy. The present study was planned to find out the incidence of prediabetes and NODM and possible mechanism of action.Methods: This was a prospective, cross‑sectional study carried out at the Department of General Medicine for a period of one and half year between August 2017 and February 2019. Normoglycemic patients whose fasting blood glucose levels below 100 mg/dL and at least one year of treatment with statins were recruited in the study. Glycaemic status, development of prediabetes and NODM and insulin resistance were the primary outcomes whereas lipid profile, adverse drug effects of statins were secondary outcomes. Collected data was analysed by suitable statistical methods.Results: A total of 146 patients were recruited and 120 completed the entire study. Mean fasting blood glucose levels before initiation of statin therapy was 89.45±10.21. After one year of statin therapy, patients were separated as prediabetics and new onset diabetics and there mean fasting blood glucose levels were 116.24±12.86 (n=10) and 152.44±20.12 (n=12) respectively. A total of 12 (10.0%) patients were developed NODM and 10 (8.2%) patients developed prediabetes at the end of statin therapy. Atorvastatin 40mg was most frequency prescribed statin followed by Atorvastatin 20mg. A total of 70 (58.3%) study participants developed mild to moderate drug related adverse effects (ADRs), statin‑induced myalgia (55.7%) was the most common ADR.Conclusions: Patients treatment with statins had developed prediabetes and NODM. Atorvastatin 40mg and greater dose significantly induced NODM. Fasting blood glucose levels should be measured periodically with prescription contains higher doses of statins

A prospective open-label randomized comparative study in Alzheimer’s disease between two commonly used drugs in coastal Indian population.

Background: Currently, therapy for Alzheimer’s disease (AD) is only symptomatic. Only two classes of drugs are approved by the United States Food and Drug Administration. Our study aimed at comparing effi cacy and safety of memantine and donepezil in moderate to severe AD patients. Methods: Totally, 22 patients with moderate to severe AD were randomized into the 2 arms of the study. The study was divided into an initial 4 weeks for determination of onset of effi cacy and subsequent 28 weeks of the treatment phase. Onset of effi cacy and response was defi ned as >20% and >50% reduction in the mean total score of functional dementia scale (FDS) and clinical global impression scale (CGIS) from baseline to the study end, respectively. Results: Onset of effi cacy on FDS and CGIS was 16.7% (mean-time 61.25 days) and 80% (mean-time 36 days) with memantine and donepezil, respectively. Response was 89.3% and 40% with memantine and Donepezil, respectively. Total reduction in FDS and CGIS score of from baseline to the study end was 39.50, 40.00, and 25.60, 27.20 with memantine and donepezil, respectively. Tolerability was 86.33% and 20% with memantine and donepezil, respectively. Anorexia, muscle cramps, constipation, headache, and insomnia, were the common side-effects and self-limiting. Safety was 100% in both groups. Conclusions: Onset of effi cacy was faster with donepezil seen at 2 weeks. Response, improvement in CGIS, FDS, and tolerability were better seen with memantine at 40 weeks. Thus, in similar clinical settings, memantine can be preferred.

Diagnosis and Management of Chronic Cough due to Extrapulmonary Etiologies.

Cough remains one of the chief complaints for which patients seek medical attention. It has been estimated that at least 25% of chronic cough cases are caused by multiple, simultaneous causes. Cough is also an important presenting feature of a number of extrapulmonary conditions, such as cardiac diseases, upper airway diseases, gastroesophageal reflux diseases and neurological/ psychological conditions. The diagnosis of extrapulmonary causes of cough is quite challenging and the management is complicated by poor treatment response if underlying etiological mechanisms are overlooked. The intention of this article is to highlight the importance and consequences of nonpulmonary etiologies of cough such as drug-induced vocal cord dysfunction, gastroesophageal reflux disease, laryngeal reflux and neuropathic disorders and to discuss about management strategies for each etiology.

Estimation of Fluoride content in ground growth green vegetables leaves (amaranths) at surrounding villages of nalgonda district Andhra Pradesh by using kit colour comparison method.

Fluoride is found to be an important trace element playing a key role in the dental and skeletal formation in humans. It is generally believed that including human being consume fluoride largely from drinking water sources and the total daily intake of fluoride by individuals from water and other sources varies from place to place depending on the amount of fluoride availability in water and other sources including food. Earlier it was believed that food was not a rich source of fluoride for humans but it is now well documented that certain types of food can have high fluoride content. Therefore the present research work is to study the fluoride content in leafy vegetable samples(amaranthus) collected from five different gram panchayat’s(villages) of Nalgonda district by using kit colour comparison method and report the fluoride content in the collected samples. finally it was found that the fluoride content in all the samples were found to ranging from 2.20-2.85mg/kg which is considered to be under safe zone(0.3-2.8mg/kg) and consumption of this leafy vegetables will not affect the humans.

Butea frondosa as a gastro protective against induced gastric lesions

The study was undertaken to evaluate the potential of alcoholic extract of Butea frondosa Koen. ex Roxb against gastric lesions induced in rats using acetic acid plus pyloric ligation. The impact of alcoholic extract of Butea frondosa in doses of 100, 200 and 400 mg/kg as single dose schedules and 200 mg/kg for 7, 14, 21 and 28 days was determined. Parameters assessed were ulcer index, total acidity, acid volume, total protein and pH, non protein sulfhydryls and gastric wall mucus. Extract in dose of 400 mg/kg as a single schedule signifi cantly reduced ulcer severity, total protein and pH as against the control (p<0.05). Concurrent fi ndings were also observed with 200 mg/kg administered for 21 and 28 days. Treatment with 400 mg/kg of the extract as a single dose and 200 mg/kg for 28 days produced an elevation in the content on non protein sulfhydryls. Gastric wall mucus was enhanced with 200 mg/kg of the extract administered for 28 days with a value of 186±2.74 ìg Alcian blue/g wet weight compared with the control (p<0.05). A palpable decline in incidence of ulcers was observed with the extract which might be largely due to the presence of fl avonoids.

Cultivation, phytochemistry, pharmacological actions and therapeutic applications of erythrina indica. Lam.

Medicinal plants are the nature’s gift to human being to make disease free healthy life. It plays a vital role to preserve our health. In our country more than 2000 medicinal plants are recognized. Erythrina indica (Fabaceae) is one of the important medicinal plants of coasts of India and Malaysia. Some of its medicinal usage has been mentioned in traditional system of medicine such as ayurveda, siddha and unani. This review attempts to encompass the available literature of Erythrina indica with respect to traditional uses, phytochemistry and summary of its pharmacological activities and clinical effects. Other aspects such as toxicity are mentioned.

Comparative evaluation of efficacy and safety profile of three anti-tuberculous regimens in Mangalore.

The aim of our study was to evaluate and compare the therapeutic efficacy & safety profile of three different antituberculous regimens for pulmonary tuberculosis. The study sample size included 90 newly diagnosed, sputum positive patients of pulmonary. tuberculosis. 30 each from different groups. The parameters studied were, therapeutic efficacy included weight gain, cough, sputum examination and safety profile: nausea, vomiting, anorexia, gastritis, hepatitis, jaundice diarrhoea, rashes, dizziness, tingling & numbness, flu like symptoms & joint aches. Group-I showed statistically significant weight gain when compared to Group-II. Improvement in cough and conversion to smear negative were seen in 100% of patients in Group-I, 83.3% of patients in Group-II and 93.3% of patients in Group-III. Therapeutic efficacy was highest with Group I regimen, followed by Group III and Group II which was least efficacious. Group II also registered; the maximum cost and highest incidence of adverse effects.

Nutritional neglect and physical abuse in children of alcoholics.

OBJECTIVE: Children of alcoholics are at higher risk for physical injuries and malnutrition. In this study nutritional neglect and physical abuse in children of alcoholics has been compared with children of non-alcoholics in a General Hospital attached to a Medical College. METHODS: 72 children of alcoholics were compared with 81 normal controls. 86.1% in case group and 49.4% in control group had malnourishment of various grades. RESULTS: 45% of the children of alcoholics had physical injuries in contrast to 23.4% of children of non-alcoholics. Malnutrition co-existed in almost all injured children of alcoholics. CONCLUSION: Alcoholism in parents of malnourished and injured children are frequently ignored in routine clinical evaluation. If the risk children are identified early some of the morbidity can be avoided.

Purification and characterization of dihydrofolate reductase from Lactobacillus leichmannii.

Dihydrofolate reductase (DHFR) (5,6,7,8-THF: NADDP+ oxidoreductase, EC 1.5.1.3) was purified 205-fold to apparent homogeneity from the crude extracts of Lactobacillus leichmannii. It has UV absorption maxima at 280 nm, M(r) of 20,000, Stokes radius of 0.34 nm and a S20.w value of 0.12 S. The preparation showed the presence of 168 amino acid residues with threonine and lysine as the NH2- and COOH- terminal end-groups respectively and a single reactive sulfhydryl group. pCMB inhibited the enzyme activity (IC50 = 2 microM). The enzyme has a pH optimum of 7.4 and is thermally inactivated at > 35 degrees C. It is activated by 0.1 M KCl and KI and 2 M urea. 3-4 M urea completely inactivated the enzyme. Enzyme has Km values of 3.5 microM and 6.2 microM for NADPH and DHF respectively, and a Ki value of 7 nM for MTX, the inhibition being competitive.

Inhibition of thymidylate synthase by pergularinine, tylophorinidine and deoxytubulosine.

The activity of thymidylate synthase (TS) purified in our laboratory from Lactobacillus leichmannii was inhibited by pergularinine (PGL) and tylophorinidine (TPD) and deoxytubulosine (DTB) isolated from the Indian medicinal plants Pergularia pallida and Alangium lamarckii respectively. Cytotoxicity studies showed that cell growth of L. leichmannii was inhibited (IC50 = 40-45 microM) by all the three alkaloids, the concentrations > 80-90 microM resulting in complete loss of the enzyme activity. Ki values of the enzyme calculated from Lineweaver-Burk and Dixon plots for PGL, TPD and DTB were 10 x 10(-6) M, 9 x 10(-6) M and 7 x 10(-6) M respectively. These are typed as 'non-competitive' inhibitors of TS. All the three alkaloids inhibited (IC50 = 50 microM) the elevated TS activity of leukocytes in cancer patients with clinically diagnosed chronic myelocytic leukemia (n = 10), acute lymphocytic leukemia (n = 8) and metastatic solid tumours (n = 3).

Reversible inhibition by pyridoxal 5'-phosphate and irreversible inactivation by urea and guanidine hydrochloride of thymidylate synthase from Lactobacillus leichmannii.

Pyridoxal 5'-phosphate reversibly inhibited thymidylate synthase from Lactobacillus leichmannii. The inhibition was competitive with dUMP (Ki = 1 microM) and non-competitive with 5,10-CH2-THF (Ki = 0.08 microM). Treatment of native or pCMB-treated enzyme with urea (5 M) or guanidine hydrochloride (4 M) resulted in inactivation and dissociation of the homodimer (74 kDa) into monomer (37 kDa).

Denaturation of pteroylpoly-gamma-glutamyl hydrolase from chicken liver by urea, thiourea and guanidine hydrochloride: altered catalytic properties of the enzyme activated by urea.

The effects of varying concentrations of urea, thiourea and guanidine hydrochloride on the enzyme activity and the isoenzymic polypeptide association of pteroylpoly-gamma-glutamyl hydrolase (EC 3.4.22.12) from chicken liver were studied. Incubation of the enzyme at 4 degrees C with low concentrations of the buffered (100 mM sodium acetate containing 1% ascorbate, pH 4.1) solutions of urea (0.55 M) and guanidine hydrochloride (0.05 M) resulted in stimulation (5- and 2-fold respectively) of the activity of the enzyme whereas at higher concentrations of the denaturants (6 M urea, 1 M thiourea or 2 M guanidine hydrochloride) the enzyme was completely inactivated. However, there was no enzyme activation in response to thiorea treatment. Under specific denaturing conditions the association of two isoenzymic polypeptides was studied. The 0.55 M urea- and 0.05 M guanidine hydrochloride-activated enzyme displayed its disaggregated nonidentical polypeptides I and II (M(r) = 41,000 and 17,300 respectively) on Sephadex G-100 gel filtration, SDS-PAGE and sedimentation analyses. The 8 M urea- and 3 M guanidine hydrochloride-inactivated enzyme on the other hand exhibited a single protein aggregate species of an M(r), 57,000 like the native enzyme. Both unmodified native enzyme and the pCMB-modified PtepolyGlu hydrolase responded similarly to these denaturants. The two constituent active polypeptides polyp-I and polyp-II of the heterodimeric gamma-glutamyl glutamyl hydrolase are dissociated in the presence of 0.55 M urea as evident from the PAGE analyses. Some catalytic properties of the activated enzyme were studied and compared with those of the native enzyme. The urea-activated enzyme displayed a shift in the second pH optimum of the double pH-activity profile (optima at pH 4.1 and pH 5.2) from pH 5.2 to pH 6.0. The activated enzyme has a Km value of 0.59 x 10(-6) M (Vmax, 0.10) for 5-CH3-H4PteGlu4 while the native enzyme has the Km of 0.83 x 10(-6) M (Vmax, 0.03) for this substrate. When the reaction mixtures were incubated with the urea-activated gamma-glutamyl hydrolase, a maximum stimulatory effect on the enzyme activity was observed with the bivalent metal ion Ca2+ whereas the most potent inhibitory effect was observed with the trivalent anion citrate.

Pteroyl- and tetrahydropteroylpolyglutamate effects on the catalytic activity of thymidylate synthase from Lactobacillus leichmannii: a novel method for determining gamma-glutamyl chain lengths of the folylpolyglutamates.

Tetrahydropteroylpolyglutamates with longer gamma-glutamyl chain lengths have been found to act as better cofactors than the corresponding monoglutamates for the activity of thymidylate synthase, (5, 10-CH2H4PteGlu: dUMP C-methyltransferase, EC 2.1.1.45) purified from Lactobacillus leichmannii. Contrarily, the pteroylpolyglutamates (unreduced forms) with longer gamma-glutamyl chain lengths act as powerful inhibitors of the same enzyme, the I50 being 2 microM for the tetraglutamate, and inhibition is competitive. The Km and Ki values for the synthetic folylpolyglutamates are identical to those obtained for the natural folylpolyglutamyl forms isolated from Torula yeast (Candida utilis) by the author earlier. A rapid novel method is suggested that could be conveniently used to determine the gamma-glutamyl chain lengths of the folylpolyglutamates employing the direct or indirect linear proportionality relationship observed between the number of gamma-glutamyl residues linked and the Ki and Km values of the enzyme considering the state of oxidation/reduction of the pteridine moiety and the 1-C substituents attached.

Physico-chemical properties of thymidylate synthase from Lactobacillus leichmannii: thiol groups, amino acid composition and the terminal end-groups.

Thymidylate synthase (5,10-methylenetetrahydrofolate: deoxyuridylate C-methyltransferase, EC 2.1.1.45) from Lactobacillus leichmannii was completely inactivated after 5 min of heat treatment at 55 degrees C. A remarkable synergistic effect with no loss in activity was noted when 10(-3) M dUMP was added to the enzyme before subjecting to heat treatment. The enzyme got activated in the presence of 2-mercaptoethanol (75 mM) and inhibited by pCMB (I50 = 5 microM). It had 2 free sulfhydryl groups and a single disulfide bond. The two identical subunits of the 74 kDa dimer were possibly bonded by a single disulfide linkage. It had a total of 652 amino acids with methionine as the amino-terminal and alanine as the carboxy-terminal amino acid residues. The carboxy-terminal end-group alanine was preceded by valine, lysine and proline sequentially in that order.

Ternary complex formation with 5-fluoro-2'-deoxyuridylate and the kinetic properties of thymidylate synthase from Lactobacillus leichmannii.

Lactobacillus leichmannii thymidylate synthase (5,10-CH2-H4PteGlu:dUMP C-methyltransferase, EC 2.1.1.45) forms a tight and stable covalently bonded ternary complex with the inhibitor 5-FdUMP in the presence of the cofactor 5,10-CH2-H4-PteGlu. 'Filter assay' employing the radioactive nucleotide ligand showed that 2 moles of FdUMP are bound per mole enzyme during the ternary complex formation with the L. leichmannii dTMP synthase. This is in line with our earlier observation on the Streptococcus faecium thymidylate synthase [Narasimha Rao, K & Kisliuk R L (1983), Proc Natl Acad Sci USA, 80, 916-920]. The enzyme has Km values of 6.3 x 10(-6) M, 8.2 x 10(-5) M and 1.0 x 10(-4) M for dUMP, (dl)-L-H4PteGlu and Mg2+ respectively; Vmax for dUMP, (dl)-L-H4PteGlu and Mg2+ are; 0.55, 0.5 and 1.1 respectively. It has K(i) values of 6.7 x 10(-6) M, 2.2 x 10(-6) M, 5.0 x 10(-5) M and 2.0 x 10(-4) M for FdUMP, dTMP, MTX and Ca2+ respectively. The type of enzyme inhibition with FdUMP, dTMP, MTX and Ca2+ was competitive. dTMP Studies clearly show the 'end product' inhibition of the enzyme.