Anaemia in Newly Diagnosed Patients of Rheumatoid Arthritis and its Correlation with Disease Activity.

Aim: To detect frequency of anemia in patients of Rheumatoid arthritis (RA) and to establish relationship between hemoglobin level and disease activity in RA. Method: Fifty nine patients of RA fulfilling 2010 ACR/EULAR criteria of RA having disease duration less than two years were included in the study. Haemoglobin (Hb) levels were measured. Disease activity was assessed by DAS-28 score. Results: Among 40/59 (67.80%) anemic cases, 22/40 (55%) patients had anaemia of chronic disease (ACD), 11/40 (27.50%) patients had Iron deficiency anemia (IDA), 3/40 (7.5%) patients had vitamin B(12) deficiency, 1/40 (2.50%) patient had folate deficiency and 3/40 (7.50%) patients had combined IDA and vitamin B(12) deficiency. Duration of disease, rheumatoid factor positivity and occurrence of erosive disease were not significantly different among anaemic and nonanaemic patients (p>0.05 for each). Mean ESR (p>0.02) and DAS-28 (p>0.001) were statistically significantly different among anaemic and nonanaemic patients. Haemoglobin level had significant negative correlation with disease activity (DAS28) in RA cases (r -0.5533, p<0.0001). Conclusion: Anemia was seen in higher frequency in RA patients. Haemoglobin had significantly negative correlation with disease activity (DAS 28) in RA. . Conclusion: Anemia was seen in higher frequency in RA patients. Haemoglobin had significantly negative correlation with disease activity (DAS 28) in RA.

Enhanced candicidal compound production by a new soil isolate Penicillium verruculosum MKH7 under submerged fermentation.

BACKGROUND: Microorganisms are a rich source of structurally diverse secondary metabolites that exert a major impact on the control of infectious diseases and other medical conditions. The biosynthesis of these metabolites can be improved by manipulating the nutritional or environmental factors. This work evaluates the effects of fermentation parameters on the production of a lactone compound effective against Candida albicans by Penicillium verruculosum MKH7 under submerged fermentation. Design-Expert version8.0 software was used for construction of the experimental design and statistical analysis of the experimental data. RESULTS: The important factors influencing antibiotic production selected in accordance with the Plackett-Burman design were found to be initial pH, temperature, peptone, MgSO4.7H2O. Orthogonal central composite design and response surface methodology were adopted to further investigate the mutual interaction between the variables and identify the optimum values that catalyse maximum metabolite production. The determination coefficient (R2) of the fitted second order model was 0.9852. The validation experiments using optimized conditions of initial pH 7.4, temperature 27 °C, peptone 9.2 g/l and MgSO4.7H2O 0.39 g/l resulted in a significant increase (almost 7 fold from 30 to 205.5 mg/l) in the metabolite production which was in agreement with the prediction (211.24 mg/l). Stability of the compound was also assessed on the basis of its response to physical and chemical stresses. CONCLUSIONS: So far as our knowledge goes, till date there are no reports available on the production of antibiotics by Penicillium verruculosum through media optimization using RSM. Optimization not only led to a 7 fold increase in metabolite yield but the same was achieved at much lesser time (8-10 days compared to the earlier 12-15 days). The enhanced yield of the antibiotic strongly suggests that the fungus P. verruculosum MKH7 can be efficiently used for antibiotic production on a large scale.

Atrial Septal Aneurysm Presenting as Clubbing without Clinically Apparent Cyanosis.

Atrial septal aneurysm (ASA) is a localised "saccular" deformity which protrudes to the right or the left atrium or on both sides. It is a rare, but well recognised cardiac abnormality. It is usually an incidental finding or may presents as atrial arrhythmias or arterial embolism. Though it is an acyanotic congenital heart disease but it may result in significant right to left shunt and cyanosis. We describe a patient of ASA with atrial septal defect who presented with clubbing and right to left shunt without clinically apparent cyanosis.

Estimation of vitamin D levels in rheumatoid arthritis patients and its correlation with the disease activity.

AIM: To detect level of serum vitamin D in patients of Rheumatoid arthritis (RA) and to establish relationship between serum vitamin D level and disease activity in RA. METHOD: Eighty patients of RA fulfilling 1987 revised criteria of the American College of Rheumatology (ACR) of RA classification and eighty healthy controls were included in the study. 25 (OH) vitamin D levels were measured. Disease activity was assessed by DAS-28 score. RESULTS: Ninety percent of RA patients were either vitamin D deficient or insufficient while only seventy percent of healthy controls were either vitamin D deficient or insufficient(p=0.007). Mean serum vitamin D levels of RA patients was significantly low compared to healthy controls (p=0.009). Thirty-one patients had high disease activity (DAS-28 score >5.1, group A), 32 patients had moderate disease activity (DAS 28 score 3.2-5.1, group B) and 17 patients had low disease activity (DAS-28 score <3.2, group C). Vitamin D levels in high disease activity group was significantly low compared to vitamin D level in patients with low and moderate disease activity (p<.001) and vitamin D level had significant negative correlation with DAS28 score (r=-0.604, p<0.001). CONCLUSION: Serum vitamin D levels were significantly low in RA patients than in healthy controls. Vitamin D deficiency was seen in significantly higher numbers of patients and vitamin D had negative correlation with disease activity in RA.

Methaemoglobinaemia as a result of nitrite poisoning.

Methaemoglobinaemia due to nitrite poisoning is rare. Awareness of this condition in the cyanosed patient not responding to oxygenation and timely administration of methylene blue may be life saving. We report a case of methaemoglobinaemia as a result of sodium nitrite poisoning.

Comparative study of antibiofilm activity of copper oxide and iron oxide nanoparticles against multidrug resistant biofilm forming uropathogens.

The effectiveness of the metal oxide nanoparticles viz. CuO and Fe2O3 as antibacterial agents against multidrug resistant biofilm forming bacteria was evaluated. CuO nanoparticles were also experimented for antibiofilm and time kill assay. The CuO displayed maximum antibacterial activity with zone of inhibition of (22 ± 1) mm against methicillin resistant Staphylococcus aureus (MRSA) followed by Escherichia coli (18 ± 1) mm. The Fe2O3 showed the zone of inhibition against MRSA of (14 ± 1) mm followed by E. coli (12 ± 1) mm. CuO proved to be more toxic than Fe2O3 nanoparticles showing significantly high antibacterial activity and found to possess dose dependent antibiofilm properties.

Anti-nucleosome antibodies in patients with systemic lupus erythematosus: potential utility as a diagnostic tool and disease activity marker and its comparison with anti-dsDNA antibody.

OBJECTIVE: To compare the utility of anti-nucleosome antibodies and anti-dsDNA antibodies in diagnosis of Systemic Lupus Erythematosus (SLE) and as a marker of disease activity. METHODS: This is a hospital based observational study among 40 (37 females and 3 males) selected cases of SLE (> or = 4 ACR criteria) and 80 control. 40 cases of other systemic autoimmune disease (SAD) [e g. 29 cases of Rheumatoid arthritis, 4 cases of Systemic sclerosis/scleroderma, 4 cases of Sjögren syndrome, 3 cases of MCTD and 40 Healthy blood were taken as control. From each patient venous blood samples were collected and submitted for anti-nucleosome and anti-dsDNA antibodies assay by enzyme linked immunosorbent assay (ELISA). RESULTS: Anti-nucleosome antibodies were positive in 19 (47.5%) SLE, 02 (05%) other SAD and none of the healthy persons. Anti dsDNA antibodies were positive in 15 (37.5%) SLE patients, 07 (17.5%) other SAD and 01(2.5%) healthy persons. For diagnosis of SLE, sensitivity of anti-ds DNA and anti-nucleosome antibody was found to be 37.5% and 47.50% respectively. The specificity of anti-nucleosome was 100% and that of anti-dsDNA was 97.50%. So, anti-nucleosome antibody test is more specific and more sensitive for diagnosis of SLE than anti-dsDNA. When SLE cases were compared with SAD, sensitivity of anti-dsDNA and anti-nucleosome antibody, for diagnosis of SLE, found to be 37.50% and 47.50% respectively but the specificity of anti-nucleosome was 95% and that of anti-dsDNA was 82.50%. Both antibodies show positive correlation with SLEDAI score .The correlation coefficient was stronger for anti-dsDNA antibodies (r = +0.550, P = < .001) than anti-nucleosome antibodies (r = +0.332, P = < .05) CONCLUSIONS: Anti-nucleosome antibodies show higher positivity than anti-dsDNA antibodies among SLE than other SAD and healthy population. Anti-nucleosome antibodies are more sensitive and specific for the diagnosis of SLE than anti-dsDNA antibodies. Anti-nucleosome and anti-dsDNA both show positive correlation with SLEDAI. But anti-dsDNA antibodies show stronger correlation with SLEDAI than anti-nucleosome. So, anti-nucleosome antibodies can be used as an additional marker for diagnosis of SLE and SLE disease activity.

Pseudo-aneurysm of left ventricle.

A 16 years boy who presented with pain chest, precordial bulging with abnormal chest movement and fever was diagnosed to have pseudo-aneurysm of left ventricle with infective endocarditis.

A metagenomic approach to decipher the indigenous microbial communities of arsenic contaminated groundwater of Assam.

Metagenomic approach was used to understand the structural and functional diversity present in arsenic contaminated groundwater of the Ganges Brahmaputra Delta aquifer system. A metagene dataset (coded as TTGW1) of 89,171 sequences (totaling 125,449,864 base pairs) with an average length of 1406 bps was annotated. About 74,478 sequences containing 101,948 predicted protein coding regions passed the quality control. Taxonomical classification revealed abundance of bacteria that accounted for 98.3% of the microbial population of the metagenome. Eukaryota had an abundance of 1.1% followed by archea that showed 0.4% abundance. In phylum based classification, Proteobacteria was dominant (62.6%) followed by Bacteroidetes (11.7%), Planctomycetes (7.7%), Verrucomicrobia (5.6%), Actinobacteria (3.7%) and Firmicutes (1.9%). The Clusters of Orthologous Groups (COGs) analysis indicated that the protein regulating the metabolic functions constituted a high percentage (18,199 reads; 39.3%) of the whole metagenome followed by the proteins regulating the cellular processes (22.3%). About 0.07% sequences of the whole metagenome were related to genes coding for arsenic resistant mechanisms. Nearly 50% sequences of these coded for the arsenate reductase enzyme (EC. 1.20.4.1), the dominant enzyme of ars operon. Proteins associated with iron acquisition and metabolism were coded by 2% of the metagenome as revealed through SEED analysis. Our study reveals the microbial diversity and provides an insight into the functional aspect of the genes that might play crucial role in arsenic geocycle in contaminated ground water of Assam.

Partial HPRT deficiency (Kelley-Seegmiller syndrome).

Hypoxanthine-guanine phosphoribosyl transferase (HPRT) deficiency is an X-linked defect of purine metabolism. Clinical manifestations are usually related to the degree of enzyme deficiency; complete HPRT deficiency (Lesh-Nyhan Syndrome) presenting with severe neurological or renal symptoms, or partial HPRT deficiency (Kelley-Seegmiller syndrome) manifesting as a gout-urolithiasis syndrome. We report a case of partial HPRT deficiency presenting as chronic tophaceous gout, mental retardation, nephrolithiasis and family history suggestive of X-linked inheritance, for its rarity.

Regulation of NAD- and NADP-linked isocitrate dehydrogenase by hydrocortisone in the brain and liver of male rats of various ages.

The activity and hormonal regulation of NAD- and NADP-linked isocitrate dehydrogenase (EC 1.1.1.41 and 1.1.1.42, respectively) in the brain and liver of rats of various ages were investigated. The activity of NAD-linked isocitrate dehydrogenase of the brain was greater than cytoplasmic or mitochondrial NADP-linked isocitrate dehydrogenase. In contrast, the cytoplasmic NADP-isocitrate dehydrogenase of the liver predominates over both NAD- and mitochondrial NADP-isocitrate dehydrogenases at the three ages studied. The activity of NAD-isocitrate dehydrogenase increased in the brain (139%) and liver (17%) of rats up to 33 weeks of age and decreased (57 and 39%, respectively) in old rats (85-week-old). The activity of cytoplasmic NADP-isocitrate dehydrogenase was maximum in immature (6-week-old) rat brain and decreased as the age of the rats increased; whereas, in liver, the activity of this enzyme was found o be maximum in adult rats (33-week-old). Brain mitochondrial NADP-isocitrate dehydrogenase activity increased (64%) in adult rats, but in liver it decreased (45 and 33% in 33- and 85-week-old rats, respectively). In both tissues, adrenalectomy and hydrocortisone treatment showed differential age-dependent response. Hydrocortisone-mediated induction of the level of enzymes was inhibited by actinomycin D.

An experimental model of oculorespiratory reflex.

An experimental study of the oculorespiratory reflex (ORR) was conducted on 20 albino rabbits using a square wave (SW) type of stimulus. The ORR could be elicited in 100% of animals. The medial rectus was observed to be most reflexogenic for ORR. The frequency and pattern of ORR was not affected by bilateral vagotomy, intravenous atropine or glycopyrrolate, but could be completely abolished by retrobular block.

Determination of fluoxymesterone, norethandrolone, prednisolone, and prednisone in tablets by differential pulse polarography.

A differential pulse polarographic method for the determination of fluoxymesterone, norethandrolone, prednisolone, and prednisone in tablets is described. This method is more sensitive than dc polarography, and the measurement of diffusion current is greatly simplified. Sørensen phosphate buffer, pH 5.6, was used as the supporting electrolyte. No apparent interference was observed from tablet excipients; the method is rapid, simple, and relatively precise.

Determination of flucytosine in tablets by differential pulse polarography.

A differential pulse polarographic assay was developed for determining flucytosine in tablets. The drug is extracted from the sample with water and hydrochloric acid and, after the pH is adjusted, an aliquot is added to the cell and the solution is polarographed at the dropping mercury electrode versus the saturated calomel electrode with 0.066 M Sørensen phosphate buffer (pH 5.6) as the supporting electrolyte. The polarographic peak height enables precise quantitative determination. The Ep value for flucytosine is--1.54 v versus the saturated calomel electrode. The mean recovery of the drug is 101.5% +/- 1.9 (SD). The method is simple, rapid, and precise.

Philosophic basis of biomedical research. I. Scientific hypotheses.

Hypotheses are central in scientific research. This has been so in all the major scientific works of our time. However, the dangers of projecting the desires of hypothesizing individual into his observations and inferences should be avoided and one should consider a hypothesis as a tentative proposition having varifiable consequences. Our present biomedical literature needs more emphasis on well conceived hypotheses. Many of these when subjected to experimentation would be proved wrong, as many have been in the past. But even these would stimulate experimental testing and eventually add to our understanding.

A new cardenolide from the roots of Terminalia arjuna.

A new cardenolide, 16,17-dihydroneridienone 3-O-beta-D-glucopyranosyl-(1-->6)-O-beta-D-galactopyranoside (1), was isolated from the roots of Terminalia arjuna.

Total serum levels of triiodothyronine (T3) thyroxine (T4) and thyrotropine (TSH) in school going children of Dibrugarh district: an endemic goitre region of Assam.

The Thyroid Status was studied by estimating the total serum levels of T3 and T4 by radioimmunoassay (RIA) and TSH by radioimmunometric Assay (IRMA) from 635 school children (8-20 years; male 129, female 506) of the Dibrugarh district: a chronic endemic goitrous region of India. The results were compared with the control group of 147 (male 48, female 99) of healthy medical students of the same geographical area. The average values of T3 and TSH of school children were found higher and T4 lower than the control; the difference were only significant for T3 and TSH. T3/T4 ratio is more in school children than the control. The findings of low T4 and high TSH indicate that the school children (Pubertal stage) from chronic iodine deficients areas suffer from poor thyroid status; the male seemed to have been affected more than the female. As age advances the thyroid status improves in female.

Immune responses to chloroquine--sensitive and resistant populations of Plasmodium berghei in mice.

In order to elucidate the role of the host as a factor in the spread of chloroquine resistance, a study of the host's immune responses in chloroquine resistant (cqr) and chloroquine sensitive (cqs) Plasmodial infections is essential. Course of the infection and the nature of immune responses in mice infected with chloroquine resistant (R) and chloroquine sensitive (S) strains of Plasmodium berghei were compared. Crude parasite antigen activated T cells from both the groups of mice (R and S) and the parasite specific antibodies were detected in the sera of both the groups. The differences in immune responses between the control (uninfected) and infected mice were found to be significant. However, humoral and in vitro cellular responses obtained with T cells from chloroquine resistant P. berghei primed mice was lower in comparison with the responses obtained with T cells from the sensitive infections. Our studies therefore suggest that immunosuppression to parasite antigen is seen in mice primed with chloroquine resistant P. berghei, which may play a role in the development of resistance.

Insight into structural organization and protein-protein interaction of non structural 3 (NS3) proteins from dengue serotypes.

Dengue infections produce a distinct character of virus-induced intracellular membrane alterations which are associated with the viral replication machinery. Currently, the NS3 protein is being targeted for antiviral therapy against dengue. NS3 protein of dengue virus interacts with nuclear receptor binding protein (NRBP) of human causing cell trafficking between the Endoplasmic Reticulum (ER) and Golgi, which interacts with Rac3, a member of the Rho-GTPase family. No crystal structure of the NRBP is available for any species, thus limiting the complete understanding of structure- function relationships of this protein. The present study deals with the molecular modeling of the viral protein (NS3 of DENV1-4), the host protein (NRBP) and their interactions through protein-protein docking study. Theoretical threedimensional structures of the NRBP and NS3 were modeled using the Modeller 9v8, and the evaluated models were docked using GRAMM-X to study the mode of protein-protein interaction (NRBP as receptor and NS3 as ligand). The docked docking complexes were further evaluated for interaction analysis by the RosettaDock Server. Suface and interface residues were observed along with hydrogen and hydrophobic interaction. The conserved residues forming hydrogen interaction of NRBP with DENV1-4 serotypes were found to be GLN 305, SER 363 and GLN 379.

Immunogenic potential and protective efficacy of formalin inactivated circulating Indian strain of West Nile virus.

OBJECTIVE: To assess the best suitable condition for virus inactivation, and to study the immunogenic potential and protective efficacy of a circulating West Nile virus (WNV) strain in Assam. METHODS: Bulk preparation of circulating WNV: WNIRGC07 (GeneBank ID: HQ246154), was undertaken in a bioreactor using cytodex-1. Virus Inactivation was done in three different conditions; 22 °C, 4 °C and room temperature. The virus preparations were evaluated for antigenicity by ELISA and toxicity by cell proliferation kit. Virus efficacy was done in-vivo on swiss albino mice against standard Indian WNV and Japanese encephalitis virus (JEV) strain. Humoral and cell mediated immune response was evaluated in mice sera by ELISA and neutralization assay. RESULTS: Inactivation at 22 °C was found to be more suitable in terms of less toxicity and high antigenicity. The same was selected to study the immune response and efficacy in mice. It induced neutralizing antibody titre of 1: 625 and high IgG response. In vivo experiment showed 100% protective efficacy against WNV and 20.8% cross protective efficacy against JEV. Further assessment of cellular immunity through immunized mice revealed augmentation of high levels of pro-inflammatory cytokines and moderate levels of anti-cytokines indicating a mixed balance of Th1 and Th2 response. CONCLUSIONS: Findings suggest that formalin inactivated Indian WNV strain has a good immunogenic potential. This is the first study on assessment of immunogenic potential of a lineage 5 strain of WNV. Our study reveals that it would be a promising and effective candidate for vaccine studies which warrants further evaluation.