Characterization of a novel low molecular weight sucrase from filamentous fungus Termitomyces clypeatus

An extracellular sucrase from the culture filtrate of filamentous basidiomycota Termitomyces clypeatus grown on high sucrose (5%, w/v) was purified by gel filtration chromatography, ion exchange chromatography and HPGPLC. The biochemical properties, molecular weight and conformation of sucrase produced were significantly different from the sucrase earlier purified from sucrose (1%, w/v) medium in the fungus. Purified sucrase was characterized as a low molecular weight protein of 13.5 kDa as approximated by SDS-PAGE and HPGPLC and exhibited predominantly random coil conformation in far-UV CD spectra. The enzyme was optimally active at 47 °C and pH 5.0. K_m and catalytic activity of the enzyme for sucrose were found to be 3.5 mM and 1.06 U/mg/mM, respectively. The enzyme was maximally active towards sucrose than to raffinose and sucrase activity was significantly inhibited by bivalent metal ions and reducing group agents. The results indicated that due to changes in aggregation pattern, molecular organization of purified sucrase, produced in high sucrose medium, was altered and was different from the previously reported enzyme. This is the first report of a sucrase of such low size showing activity.     

Effects of different feeding levels on moulting,growth and conversion efficiency of Palaemon lamarrei

Pigment analyses by TLC were carried out during and after the peak of a Peridinium bloom in Lake Kinneret (Israel). During the experimental period considerable changes occurred in the absolute and relative amounts of the various pigments, especially carotene, diadinoxanthine, peridinin, chlorophyll a and chlorophyl c. The ratio between total Carotenoid to total chlorophyll content is also followed. Our findings show that considerable changes in pigment composition in natural phytoplankton can occur while the species composition remains constant.     

Cajanus cajan shows multiple novel adaptations in response to regular mechanical stress

Journal of Plant Research - Cajanus cajan is one of the least studied crop plants regarding its responses to stress conditions. Regular mechanical stress suppresses plant physiology and growth at...     

Enhanced activity and stability of cellobiase (β-glucosidase: EC 3.2.1.21) produced in the presence of 2-deoxy-d-glucose from the fungus Termitomyces clypeatus

Generally less glycosylation or deglycosylation has a detrimental effect on enzyme activity and stability. Increased production and secretion of cellobiase was earlier obtained in the presence of the glycosylation inhibitor 2-deoxy-d-glucose in filamentous fungus Termitomyces clypeatus [Mukherjee, S.; Chowdhury, S.; Ghorai, S.; Pal, S.; Khowala, S. Biotechnol. Lett.2006, 28, 1773-1778]. In this study the enzyme was purified from the culture medium by ultrafiltration and gel-permeation, ion-exchange and high-performance liquid chromatography, and its catalytic activity was six times higher compared to the control enzyme. K_m and V_max of the purified enzyme were measured as 0.187 mM and 0.018 U mg⁻¹, respectively, using pNPG as the substrate. The enzyme had temperature and pH optima at 45 °C and pH 5.4, respectively, and retained full activity in a pH range of 5-8 and temperatures of 30-60 °C. Interestingly less glycosylated cellobiase was resistant towards proteolytic as well as endoglycosidase-H digestion and showed higher stability than native enzyme due to increased aggregation of the protein. The enzyme also showed higher specific activity in the presence of cellobiose and pNPG and less susceptibility towards salts and different chemical agents. The β-glucosidase can be considered as a potentially useful enzyme in various food-processing, pharmaceutical and fermentation industries.     

Cellobiase from Termitomyces clypeatus: activity and secretion in presence of glycosylation inhibitors

In presence of the glycosylation inhibitors, 2-deoxy-d-glucose (1 mg/ml), tunicamycin (30 μg/ml), 1-deoxynojirimycin (30 μg/ml) and d-glucono-δ-lactone (1 mg/ml), total cellobiase activity, in the extracellular, intracellular and cell bound fractions, of the fungus Termitomyces clypeatus grown in 20 ml cellobiose medium (1%, w/v) increased by 50-, 1.8-, 2.4-, 1.3-fold, respectively, with respect to control medium (16.3 U). The inhibitors also stimulated secretion of 95% of the total protein in culture medium, except d-glucono-δ-lactone which released 60% of the total protein. 2-Deoxy-d-glucose (1 mg/ml) led to production of extracellular cellobiase up to 40 U/ml, whereas in absence of the inhibitors only 0.59 U/ml enzyme was detected.     

Alteration of testicular biochemistry during protein restriction in nickel treated rats

Nickel sulfate (2.0 mg/100 g.b.wt) dissolved in double-distilled water was administered (ip) on alternate days for ten doses to normal protein-fed and protein-restricted Wister strain albino rats (b.wt. 160 ± 5 g). Two groups were used: one with normal protein diet, whereas the other with protein-restricted diet served as control. Twenty-four hours after the last treatment, the animals were sacrificed by decapitation. Tissues such as the testes, seminal vesicles, epididymis (Cauda and Caput) and prostate were dissected out, wiped clean, and stored at -20‡C until analysis. Lactate dehydrogenase (LDH) activities, glutamate oxaloacetate transaminase (GOT) activities, glycogen content, cholesterol content, and total protein content of the testes were estimated. Nickel sulfate administration significantly decreased the body weight of both normal protein-fed and proteinrestricted groups of animals; the organ weights were also decreased. Significant decrease of LDH activity was observed, but GOT activity was not altered significantly. Testicular glycogen and cholesterol increased significantly in both experimental groups, but total protein content decreased. Nickel sulfate seems to have an adverse effect on the male reproductive system in both groups of animals fed with normal protein (18% casein) diet and protein restricted (5% casein) diet.     

Effects of eyestalk ablation on growth and food conversion efficiency of the freshwater prawn Macrobrachium lanchesteri (de Man)

Bilateral eyestalk ablation in the freshwater prawn Macrobrachium lanchesteri results in high mortality, while unilateral eyestalk ablated prawns exhibited a high survival rate. There was marked increase in the growth of bilateral eyestalk-ablated prawns (47.70 mg/prawn) as compared to those that were unilaterally ablated (19.19 mg/prawn).     

Factors associated with delays in treatment initiation after tuberculosis diagnosis in two districts of India

Background

Excessive time between diagnosis and initiation of tuberculosis (TB) treatment contributes to ongoing TB transmission and should be minimized. In India, Revised National TB Control Programme (RNTCP) focuses on indicator start of treatment within 7 days of diagnosis for patients with sputum smear-positive PTB for monitoring DOTS implementation.

Objectives

To determine length of time between diagnosis and initiation of treatment and factors associated with delays of more than 7 days in smear-positive pulmonary TB.

Methods

Using existing programme records such as the TB Register, treatment cards, and the laboratory register, we conducted a retrospective cohort study of all patients with smear-positive pulmonary TB registered from July-September 2010 in two districts in India. A random sample of patients with pulmonary TB who experienced treatment delay of more than 7 days was interviewed using structured questionnaire.

Results

2027 of 3411 patients registered with pulmonary TB were smear-positive. 711(35%) patients had >7 days between diagnosis and treatment and 262(13%) had delays >15 days. Mean duration between TB diagnosis and treatment initiation was 8 days (range = 0–128 days). Odds of treatment delay >7 days was 1.8 times more likely among those who had been previously treated (95% confidence interval [CI] 1.5–2.3) and 1.6 (95% CI 1.3–1.8) times more likely among those diagnosed in health facilities without microscopy centers. The main factors associated with a delay >7 days were: patient reluctance to start a re-treatment regimen, patients seeking second opinions, delay in transportation of drugs to the DOT centers and delay in initial home visits. To conclude, treatment delay >7 days was associated with a number of factors that included history of previous treatment and absence of TB diagnostic services in the local health facility. Decentralized diagnostic facilities and improved referral procedures may reduce such treatment delays.