Production of recombinant enveloped structural proteins from the Chinese WSSV isolate

The white spot syndrome virus (WSSV) is one of the deadly pathogens of penaeid shrimps and other crustaceans. The WSSV virion consists of an enveloped rod-shaped nucleocapsid enclosing a large circular double stranded DNA genome of 305 Kb with 181 open reading frames. The two major structural genes, VP19 and VP28 were amplified from the genomic DNA of Chinese isolate of WSSV and cloned in pUCm-T vector and sub cloned in pET-30a (+) vector. The expressions of genes inE. coli (BL21) were confirmed by SDS-PAGE analysis. The clones were sequenced, submitted to the gene bank and the Xiang Shan strain of WSSV were compared with the previous reported sequence of WSSV of various regions which revealed that VP19 and VP28 gene sequences had certain differences from the sequences of similar genes of the isolate already reported. The recombinant proteins expressed, purified and characterized.     

Metabolic Bone Profile of Healthy Adult North Indian Population from Chandigarh Urban Bone Epidemiological Study (CUBES)

We aimed to estimate metabolic bone profile in a large cohort of healthy, adult Indian population to generate reference standards of serum calcium, phosphate and alkaline phosphatase (ALP), 25 (OH) Vitamin D and iPTH, and also to find out the prevalence of Vitamin D deficiency in healthy population. Apparently healthy people in the age group of 20–80 years, residing in the union territory of Chandigarh were chosen. Fasting samples for serum calcium, phosphate, albumin, alkaline phosphatase (ALP), 25 (OH) D and iPTH were collected and were processed on the same day. We recruited 930 healthy subjects from different subsectors of Chandigarh. Final analysis was done for 915 subjects. Out of this, 530 (58%) were women and 385 (42%) were men. The study participants were divided into two groups, less than and more than 50 years for the men and pre and post-menopausal for the women. The serum calcium, phosphate, ALP and iPTH were significantly higher in the post-menopausal women compared to the pre-menopausal women. The median plasma 25 (OH) D in men and women was 12.5 ng/mL and 14.3 ng/mL, respectively. 25 (OH) D deficiency was seen in 65.4% of individuals. 25 (OH) D levels co-related negatively with iPTH levels (r = − 0.4, p < 0.0001), and showed an increasing trend with age. We have thus presented metabolic bone profile of healthy, adult north Indian population. These reference values can be used for diagnosis and monitoring of various MBDs. Vitamin D deficiency is still rampant in our population in spite of increasing awareness.     

Iron sulfur proteins and their synthetic analogues: Structure, reactivity and redox properties

A number of non-heme proteins contain iron-sulfur clusters as their core. The various structural types that have been characterised so far are discussed. Attempts to understand their properties and functions at a molecular level through model systems are described.     

Enzyme vs. extremozyme

Extremozymes that function in the extreme environmental conditions of temperature, pressure, pH, and salinity are of great importance for industrial processes and scientific research. Apart from understanding the principle behind the stability of such enzymes, there has been considerable effort to isolate enzymes from extremophiles (organisms that live in extreme environmental conditions) as well as modify enzymes from mesophiles (organisms that live in normal environmental conditions) to convert them into extremozymes.     

Mycobacterium tuberculosis H37Ra ESAS-7—An excretory-secretory antigen fraction of immunodiagnostic potential in pulmonary tuberculosis

A mycobacterial excretory-secretory protein fraction ESAS-7 purified by 50% ammonium sulphate precipitation followed by SDS-PAGE fractionation was evaluated by penicillinase enzyme linked immuno-sorbent assay (ELISA) for its sensitivity and specificity in the diagnosis of pulmonary tuberculosis. At a “cut off” serum dilution of 600, 38 (90%) of 42 sera from bacteriologically confirmed tuberculosis cases, 15 (100%) of 15 sera from bacteriogically negative but anti tubercular therapy (ATT) responded cases, 3 (7%) of 43 sera from normal healthy subjects and 4 (8%) of 48 sera from non tuberculous disease control cases gave positive reaction for tubercular antibody to ESAS-7 antigen fraction containing predominantly 33-kDa protein with a sensitivity of 90% in bacteriologically confirmed cases and specificity of 92%. Further, this diagnostic assay using the ESAS-7 antigen is more sensitive requiring as little as one nanogram antigen per test compared to use of 100 nanogram EST-6 antigen reported earlier. Thus use of ESAS-7 antigen for antibody detection has good diagnostic potential with improved specificity in pulmonary tuberculosis.     

Salivary proteolytic activities in periodontitis, gingivitis and diabetes mellitus

Elastase activity was found to be significantly increased in periodontitis (0.872±0.4270 absorbance units/mg protein, mean±S. D., 1.05±0.61 units/ml saliva), gingivitis (0.772±0.416 units/mg protein, 1.515±0.952 units/ml) and diabetes (0.549±0.286 units/mg protein, 1.20±0.769 units/ml) compared to normals (0.255±0.089) units/mg protein, 0.264±0.079 units/ml). Chymotryptic activity was not found to be increased in these disease conditions over the normal level (0.284±0.096 units/mg protein). Aminopeptidase activity was elevated only in periodontitis (0.670+0.140 units/mg protein) compared to normals (0.349±0.100 units/mg protein). Trypsin-like activity was also found to be significantly raised in periodontitis compared to normals when Pro-Phe-Arg-p-nitroanilide (0.666±0.204 units/mg protein), Ile-Pro-Arg-p-nitroanilide (1.59±0.260 units/mg protein) and Pyroglu-Pro-Arg-p-nitroanilide (1.82±0.380 units/mg protein) were used as substrates. The normal values with these three substrates were respectively, 0.454±0.110, 1.04±0.231 and 1.15±0.312 units/mg protein. Total protein level in saliva was found to be significantly elevated in gingivitis (209±66.8 mg/dl) and diabetes (204±68.0) compared to normal values (107±20.7). Increase in periodontitis was marginal (127±28.3 mg/dl). Expression of proteolytic activities normalized to protein level was useful in differential diagnosis. Increase in elastase level in saliva is not a specific marker for periodontal diseases.     

Preliminary studies on serum lipids, apolipoprotein-B and oxidative stress in xanthelasma

Serum lipid profile, apolipoprotein-B (apo-B), malondialdehyde levels(MDA) and superoxide dismutase (SOD) activity were assessed in 12 cases of xanthelasma with and without coronary artery disease (CAD)/hypertension (HTN) and results are compared with healthy controls. Dyslipidemia was found in 65% cases of xanthelasma as compared to 20% healthy controls. Xanthelasma patients had significantly high malondialdehyde (MDA) levels (p<0.01) and significantly decreased (p<0.05) SOD activity as compared to controls. Among xanthelasma patients, xanthelasma with CAD/HTN showed higher total cholesterol (236±32.7 vs 188±24.7 mg/dl), low density lipoprotein cholesterol (157±35.5 vs 113±16 mg/dl) and Apo-B (120.5±9.4 vs 114±19.2 mg/dl) levels as compared to xanthelasma without CAD/HTN. Results of our study indicate that xanthelasma patients with increased apo-B, MDA and decreased SOD need cardiovascular monitoring.