Comparative structural and enzymatic studies on Salmonella typhimurium diaminopropionate ammonia lyase reveal its unique features.

Cellular metabolism of amino acids is controlled by a large number of pyridoxal 5'-phosphate (PLP) dependent enzymes. Diaminopropionate ammonia lyase (DAPAL), a fold type II PLP-dependent enzyme, degrades both the D and L forms of diaminopropionic acid (DAP) to pyruvate and ammonia. Earlier studies on the Escherichia coli DAPAL (EcDAPAL) had suggested that a disulfide bond located close to the active site may be crucial for maintaining the geometry of the substrate entry channel and the active site. In order to obtain further insights into the catalytic properties of DAPAL, structural and functional studies on Salmonella typhimurium DAPAL (StDAPAL) were initiated. The three-dimensional X-ray crystal structure of StDAPAL was determined at 2.5 Šresolution. As expected, the polypeptide fold and dimeric organization of StDAPAL is similar to those of EcDAPAL. A phosphate group was located in the active site of StDAPAL and expulsion of this phosphate is probably essential to bring Asp125 to a conformation suitable for proton abstraction from the substrate (D-DAP). The unique disulfide bond of EcDAPAL was absent in StDAPAL, although the enzyme displayed comparable catalytic activity. Site directed mutagenesis of the cysteine residues involved in disulfide bond formation in EcDAPAL followed by functional and biophysical studies further confirmed that the disulfide bond is not necessary either for substrate binding or for catalysis. The activity of StDAPAL but not EcDAPAL was enhanced by monovalent cations suggesting subtle differences in the active site geometries of these two closely related enzymes.

Unusual direct immunofluorescence (DIF) pattern in pemphigus herpetiformis- A case report.

Pemphigus herpetiformis (PH) is an autoimmune intraepithelial bullous skin disorder. A 61-year-old female presented with history of multiple pruritic erosions, ulcers all over body, and diffuse loss of hair over scalp. Oral and genital mucosas were uninvolved. Subcorneal separation with suprapapillary thinning of epidermis, neutrophilic spongiosis, and elongation of rete ridges were seen on histopathology. Direct immunofluorescence (DIF) revealed IgG deposits in intercellular zone in fish net like pattern and focal linear IgA deposits along basement zone. Indirect immunofluorescence (IIF) revealed antibodies to desmoglein1 (Dsg-1) positive. A final diagnosis of PH was given. The patient responded well to treatment with dapsone and steroids.

Hedgehog inhibition prolongs survival in a genetically engineered mouse model of pancreatic cancer.

BACKGROUND AND AIMS: Pancreatic cancer is among the most dismal of human malignancies. Current therapeutic strategies are virtually ineffective in controlling advanced, metastatic disease. Recent evidence suggests that the Hedgehog signalling pathway is aberrantly reactivated in the majority of pancreatic cancers, and that Hedgehog blockade has the potential to prevent disease progression and metastatic spread. METHODS: Here it is shown that the Hedgehog pathway is activated in the Pdx1-Cre;LsL-Kras(G12D);Ink4a/Arf(lox/lox) transgenic mouse model of pancreatic cancer. The effect of Hedgehog pathway inhibition on survival was determined by continuous application of the small molecule cyclopamine, a smoothened antagonist. Microarray analysis was performed on non-malignant human pancreatic ductal cells overexpressing Gli1 in order to screen for downstream Hedgehog target genes likely to be involved in pancreatic cancer progression. RESULTS: Hedgehog inhibition with cyclopamine significantly prolonged median survival in the transgenic mouse model used here (67 vs 61 days; p = 0.026). In vitro data indicated that Hedgehog activation might at least in part be ascribed to oncogenic Kras signalling. Microarray analysis identified 26 potential Hedgehog target genes that had previously been found to be overexpressed in pancreatic cancer. Five of them, BIRC3, COL11A1, NNMT, PLAU and TGM2, had been described as upregulated in more than one global gene expression analysis before. CONCLUSION: This study provides another line of evidence that Hedgehog signalling is a valid target for the development of novel therapeutics for pancreatic cancer that might be worth evaluating soon in a clinical setting.

Utilization of various carbon sources for the growth of waterborne conidial fungi.

Four isolates of waterborne conidial fungi (Tetracheatum elegans, Tetracladium marchalianum, Pestalotiopsis submersus and Flagellospora penicillioides) were investigated for their carbon requirement, using eight different carbon sources (viz. glucose, fructose, sucrose, xylose, starch, cellulose, dextrin and lactose). All fungi tested grew sparsely on the basal medium lacking in carbon, which was the control. However these fungi were found to vary in their ability to use the supplied sources of carbon. Glucose and sucrose were found to be suitable sources of carbon for all four fungal isolates, whereas fructose proved good for T. marchalianum and P. submersus. Starch and xylose also supported growth of T. marchalianum, P. submersus and F. penicillioides. Cellulose, a polysaccharide, was a poor source of carbon for the growth of these isolates. Four g/L of glucose was recorded as the most useful concentration that gives the maximum dry weight of selected fungi (262 mg and 400 mg for T. elegans and P. submersus respectively after 15 d).

Zinc Requirement for Stomatal Opening in Cauliflower.

Zn deficiency induced increases in epicuticular wax deposits, lamina thickness, degree of succulence, water saturation deficit, diffusive resistance, and proline accumulation and decreases in carbonic anhydrase activity, water potential, stomatal aperture, and transpiration in the leaves of cauliflower (Brassica oleracea L. var botrytis cv Pusa) plants. Restoration of Zn supply to the deficient plants increased stomatal aperture, transpiration, and carbonic anhydrase activity significantly within 2 h. However, leaf water potential in the Zn-deficient plants did not recover within 24 h after resupply of Zn. The guard cells in epidermal peels from the Zn-deficient leaves had less K+ than those from the controls. Stomatal aperture in the epidermal peels from Zn-deficient leaves was 64% less than in the controls when the epidermal strips were floated on 125 mM KCl. Supplementing the ambient medium 25 mM KCl with ZnCl2 enhanced stomatal aperture in both control and Zn-deficient peels, and the effect was significant in the latter. The observations indicate involvement of Zn in stomatal opening, possibly as a constituent of carbonic anhydrase needed for maintaining adequate [HCO3-] in the guard cells, and also as a factor affecting K+ uptake by the guard cells.

Effect of cadmium on growth, ultramorphology of leaf and secondary metabolites of Phyllanthus amarus Schum. and Thonn.

The pollution is increasing in the environment by different kinds of human activities, which results in the accumulation of heavy metals including cadmium in the soil and water and it causes different types of problems to living beings. As the plants are utilized by human being as food and medicine, therefore, it is mandatory to see the effect of metals on plants. In this context, efforts have been made to observe the effect of different concentration of Cadmium (Cd) on Phyllanthus amarus Schum. and Thonn., because Cd is the widespread metal and the plants response to low and high level of exposure is a complex phenomenon. P. amarus is mostly grown as weed in agricultural and waste lands. It is a reputed plant used in Indian indigenous systems of medicine with hepatoprotective, diuretic, stomachic properties and is recently being used for the treatment of hepatitis B. The study revealed that Cd causes significant decrease in fresh and dry weight, length of root and shoot, protein, chlorophyll, carotenoids and sugar and increase in starch content. It is interesting to note that the therapeutically active compounds-phyllanthin and hypophyllanthin, enhanced at certain levels of Cd due to abiotic stress. Besides, the ultramorpholical changes were also observed in stomatal opening and wax deposition on both the surfaces of leaves.

Adaptive radiotherapy in lung cancer: dosimetric benefits and clinical outcome.

OBJECTIVE: Anatomical changes during radiotherapy (RT) might introduce discrepancies between planned and delivered doses. This study evaluates the need for adaptive treatment in lung cancer RT. METHODS: 15 patients with non-small-cell lung cancer, undergoing radical RT with or without concurrent chemotherapy, consecutively underwent planning CT scans at baseline and after 44-46 Gy. Target volumes were delineated on both scans. Phase I delivered 44-46 Gy to the initial planning target volume (PTV). Two Phase II plans for 16-20 Gy were developed on initial and mid-treatment scans, the treatment being delivered with the mid-treatment plan. The second CT structure set was fused with the initial scan data set using dose wash. Volumetric and dosimetric changes in target volumes and critical structures were assessed. RESULTS: There was significant reduction in primary gross tumour volume (34.00%; p = 0.02) and PTV (34.70%; p < 0.01) in the second scan. In Plan 2, delivering the same dose to the initial PTV would have resulted in a significantly higher dose to the lung PTV (V20, 52.18%; V5, 21.76%; mean, 23.93%), contralateral lung (mean, 29.43%), heart (V10, 81.47%; V5, 56.62%; mean, 35.21%) and spinal cord (maximum dose, 37.53%). CONCLUSION: Treatment replanning can account for anatomical changes during RT and thereby enable better normal tissue sparing, while allowing radical target doses with the possibility of maximizing local control. ADVANCES IN KNOWLEDGE: This study supports the sparse dosimetric data regarding the quantitative tumour volume reduction, re-emphasizing the need for adaptive replanning for minimizing normal tissue toxicity without compromising local control, and adds to the existing body of literature.

Developments in chemical approaches to treat tuberculosis in the last decade.

Tuberculosis (TB) caused by Mycobacterium tuberculosis claims millions of lives each year globally. Although it can be controlled by currently available drug regimen (DOTS), yet the emergence of multidrug resistance (MDR) and extensively drug resistance (XDR) TB is a growing concern. The increasing rate of MDR-TB, co-infection with HIV and XDR-TB necessitates the development of new anti-TB agents that have a practical impact on tuberculosis control. This review article gives a brief introduction of tuberculosis, present day problems, traditional and new anti-TB drug targets, currently used drugs, their mode of action, the pipeline compounds and a short description of new chemical entities (NCE's) as antitubercular agents developed in last 10 years.

Prevalence of classical swine fever virus in India: a 6-year study (2004-2010).

A study was undertaken regarding the prevalence of classical swine fever virus (CSFV) antibodies and antigens in sera and suspected tissue samples of domestic pigs. The samples were received between January 2004 and September 2010. A total of 594 serum samples from 12 states and 287 tissue samples from 13 states of India were tested using commercial enzyme-linked immunosorbent assay (ELISA) kits. The mean prevalence of CSFV antibodies in suspected sera was 63.3% (376/594), whereas 76.7% (220/287) of the suspected samples were found to contain CSFV antigen. The high prevalence of CSFV antibodies suggests that the disease is endemic in the country. This baseline data will be of use in the formulation of control and eradication programmes.

High-performance infrared narrow-bandpass filters for the Indian National Satellite System meteorological instrument (INSAT-3D).

This paper describes the design and manufacture of a set of precision cooled (210 K) narrow-bandpass filters for the infrared imager and sounder on the Indian Space Research Organisation (ISRO) INSAT-3D meteorological satellite. We discuss the basis for the choice of multilayer coating designs and materials for 21 differing filter channels, together with their temperature-dependence, thin film deposition technologies, substrate metrology, and environmental durability performance.

Retained foreign body in sino-orbital region.

It is rare to find foreign bodies in nose and paranasal sinuses. The further rarity is to encounter impacted foreign body in this region and route of entry being through the orbit. We are here by presenting a case, where a metallic foreign body was retained for duration of 4 months in sino - orbital region.

Aetiology of pneumonia in hospitalized children.

One-hundred-and-thirty-two children with clinical and radiological evidence of bronchopneumonia/pneumonia were studied over a 1-year period for isolation/detection of bacterial and viral aetiological pathogens. Throat swab, nasopharyngeal aspirate (NPA), and lung aspirate were studied for bacterial and viral cultures. NPA was also subjected to latex agglutination test (LA) for H. influenzae and S. pneumoniae; and immunofluorescent technique (IFAT) and enzyme immunoassay (EIA) for respiratory syncytial virus (RSV). Blood culture for bacterial pathogens, and LA of blood and urine was also undertaken. Haemophilus influenzae was the commonest organism (15 per cent) isolated as the sole pathogen followed by RSV (14 per cent), Klebsiella (13 per cent) and S. pneumoniae (12 per cent). E. coli was the commonest organism (50 per cent) in infants <3 months and was closely followed by RSV (44 per cent), Klebsiella (25 per cent), and S. pneumoniae (18 per cent). Isolation rate of E. coli gradually declined with age. RSV (47 per cent) and H. influenzae (31 per cent) were the commonest organisms between 7 and 24 months. S. pneumoniae and Staph. aureus were common bacterial pathogens identified in all age groups with maximum isolation of 20 and 40 per cent, respectively, in children more than 5 years. Isolation of E. coli, Klebsiella and Staph. aureus was highest from NPA culture, while as S. pneumoniae and H. influenzae were most often detected by LA. Out of 12 cases from whom a lung aspirate was collected, bacterial pathogen could be isolated in six cases (50 per cent). Detection of RSV by EIA was higher than by culture or IFAT. Most of the organisms were resistant to chloramphenicol except for H. influenza. All the isolates of S. pneumoniae were sensitive to all the antibiotics. Bacterial pathogens were isolated/detected in 74 per cent of cases and RSV was the aetiological agent in 49 per cent of cases investigated for viral aetiology. Higher detection rate of RSV is attributed to selection of cases in winter months during a period of suspected epidemic of RSV.