Adaptability in enzymes

In studying inhibitors and activators of enzymes it has often been found that inhibitors act as activators when in very low concentrations and that certain activators (in suitable concentrations) show inhibition. Similar results were obtained in the present work in studying urease from soya bean in the presence of lead acetate. Lead acetate is normally an inhibitor of urease. It was found, however, that in small concentrations of lead acetate increase in the activity of urease occurs. On prolonging the time of the reaction inhibition develops and still later increased enzymatic activity again occurs. The results were elaborated statistically and are discussed in relation to the use of “adaptation” of urease in unfavourable conditions in reaction mixtures by the action of lead acetate.     

Recovery of nitrogen fertilizer by traditional and improved rice cultivars in the Bhutan Highlands

The recovery of soil derived nitrogen (N_DFS) and fertilizer N (N_DFF) was investigated in highland rice (Oryza sativa L.) fields in Bhutan, characterized by high inputs of farmyard manure (FYM). The effect of 60 kg N ha^?1 (60 N) applied in two splits to a traditional and an improved cultivar, popular among the farmers, was investigated using the ¹⁵N isotope dilution technique. No differences were found between cultivars with respect to the uptake of N_DFS and N_DFF, but the improved cultivar yielded 27% more (P?≤?0.05) grain compared with the traditional cultivar. This was largely due to its greater harvest index (HI). The mean percentage recovery of fertilizer N (REN) applied at 45 days after transplanting (DAT) was 34% compared to 22% at 7 DAT, resulting in 56% greater uptake of N_DFF at 45 DAT. The overall REN for both the improved and the traditional cultivars were 25.7% and 30% respectively, with no difference between cultivars, but REN decreased with increasing FYM inputs. Fertilizer N recommendations that allow for previous FYM inputs combined with applications timed to coincide with maximum crop demand (45 DAT), and the use of improved cultivars, could enhance N fertilizer recoveries (REN) and increase rice yields in the Bhutan Highlands.     

LED-Fluorescence Microscopy for Diagnosis of Pulmonary Tuberculosis under Programmatic Conditions in India

Background

Light-emitting diode fluorescence microscopy (LED-FM) has been shown to be more sensitive than conventional bright field microscopy using Ziehl-Neelsen (ZN) stain in detecting sputum smear positive tuberculosis in controlled laboratory conditions. In 2012, Auramine O staining based LED-FM replaced conventional ZN microscopy in 200 designated microscopy centres (DMC) of medical colleges operating in collaboration with India’s Revised National Tuberculosis Control Programme. We aimed to assess the impact of introduction of LED-FM services on sputum smear positive case detection under program conditions.

Methods

This was a before and after comparison study. In 15 randomly selected medical college DMCs, all presumptive TB patients who underwent sputum smear examination in the years 2011 (before LED-FM) and 2012 (after LED-FM) were compared. An additional 15 comparable DMCs that implemented conventional ZN sputum smear microscopy were also selected for comparison between 2011 and 2012.

Results

The proportion of presumptive TB patients (PTP)found sputum smear positive increased by 30%- from 13.6% (3432/25159) in 2011 to 17.8% (4706/26426) in 2012 (P value <0.01) in the sites that implemented LED-FM microscopy, whereas in DMCs where the ZN staining procedure is followed the proportion of sputum smear positive had remained unchanged (13.0%versus 12.6%;P value0.31).

Conclusion

Use of LED-FM significantly increased the proportion of smear positive cases among presumptive TB patients under routine program conditions in high workload laboratories. The study provides operational evidence needed to scale-up the use of LED-FM in similar settings in India and beyond.     

Anti-CD70 Immunocytokines for Exploitation of Interferon-γ-Induced RIP1-Dependent Necrosis in Renal Cell Carcinoma

Metastatic renal cell carcinoma (RCC) is an incurable disease in clear need of new therapeutic interventions. In early-phase clinical trials, the cytokine IFN-γ showed promise as a biotherapeutic for advanced RCC, but subsequent trials were less promising. These trials, however, focused on the indirect immunomodulatory properties of IFN-γ, and its direct anti-tumor effects, including its ability to kill tumor cells, remains mostly unexploited. We have previously shown that IFN-γ induces RIP1 kinase-dependent necrosis in cells lacking NF-κB survival signaling. RCC cells display basally-elevated NF-κB activity, and inhibiting NF-κB in these cells, for example by using the small-molecule proteasome blocker bortezomib, sensitizes them to RIP1-dependent necrotic death following exposure to IFN-γ. While these observations suggest that IFN-γ-mediated direct tumoricidal activity will have therapeutic benefit in RCC, they cannot be effectively exploited unless IFN-γ is targeted to tumor cells in vivo. Here, we describe the generation and characterization of two novel ‘immunocytokine’ chimeric proteins, in which either human or murine IFN-γ is fused to an antibody targeting the putative metastatic RCC biomarker CD70. These immunocytokines display high levels of species-specific IFN-γ activity and selective binding to CD70 on human RCC cells. Importantly, the IFN-γ immunocytokines function as well as native IFN-γ in inducing RIP1-dependent necrosis in RCC cells, when deployed in the presence of bortezomib. These results provide a foundation for the in vivo exploitation of IFN-γ-driven tumoricidal activity in RCC.     

A Prospective Epidemiological Study of Acute Mountain Sickness in Nepalese Pilgrims Ascending to High Altitude (4380 m)

Background

Each year, thousands of pilgrims travel to the Janai Purnima festival in Gosainkunda, Nepal (4380 m), ascending rapidly and often without the aid of pharmaceutical prophylaxis.

Methods

During the 2012 Janai Purnima festival, 538 subjects were recruited in Dhunche (1950 m) before ascending to Gosainkunda. Through interviews, subjects provided demographic information, ratings of AMS symptoms (Lake Louise Scores; LLS), ascent profiles, and strategies for prophylaxis.

Results

In the 491 subjects (91% follow-up rate) who were assessed upon arrival at Gosainkunda, the incidence of AMS was 34.0%. AMS was more common in females than in males (RR = 1.57; 95% CI = 1.23, 2.00), and the AMS incidence was greater in subjects >35 years compared to subjects ≤35 years (RR = 1.63; 95% CI = 1.36, 1.95). There was a greater incidence of AMS in subjects who chose to use garlic as a prophylactic compared to those who did not (RR = 1.69; 95% CI = 1.26, 2.28). Although the LLS of brothers had a moderate correlation (intraclass correlation = 0.40, p = 0.023), sibling AMS status was a weak predictor of AMS.

Conclusions

The incidence of AMS upon reaching 4380 m was 34% in a large population of Nepalese pilgrims. Sex, age, and ascent rate were significant factors in the development of AMS, and traditional Nepalese remedies were ineffective in the prevention of AMS.     

Variation in the Activity of Three Principal Detoxifying Enzymes in Major Sucking Pest of Tea, Helopeltis theivora Waterhouse (Heteroptera: Miridae) from Sub-Himalayan Tea Plantations of West Bengal, India

Helopeltis theivora Waterhouse (Heteroptera: Miridae), is a major sucking pest of tea in North East India along with other tea growing countries. In West Bengal, tea is cultivated in three sub-Himalayan regions, Terai (foothill plains western to river Teesta), the Dooars (foothill plains eastern to river Teesta) and the Darjeeling hill slopes. Most plantations, in these regions are managed conventionally i.e. by spraying different synthetic insecticides and a few by organic farming using different herbal and microbial insecticides. In conventional plantations, continuous application of insecticides may lead to the selection of more tolerant H. theivora populations making the pest difficult to control. So, there is a pressing need to know the biochemical variability in relation to the metabolic resistance in the pest populations and develop a population specific control strategy. Activity of three principal insecticide detoxifying enzymes in H. theivora populations were studied from three tea growing regions of North Bengal. Higher levels of activity of all the studied enzymes were found from conventional tea plantations. In male H. theivora, the activity of general esterases (GEs) was 6.6–11.2 and 10.5–11.4 fold higher, cytochrome P450 (CYPs) was 2.0–3.2 and 3.0–3.2 fold higher and glutathione S-transferase (GSTs) was 5.2–8.3 and 6.4–8.7 fold higher in Terai and the Dooars populations, respectively than organic populations from Darjeeling hill slopes. Similarly, in female H. theivora, activity of GEs was 6.2–10.3 and 8.3–9.6 folds higher, CYPs was 1.9–3.2 and 3.0–3.3 fold, and GSTs was 3.5–5.4 and 4.4–6.0 fold higher in Terai and the Dooars, respectively than the organic populations from Darjeeling hill slopes. The activities of all three enzymes were found to be significantly low in organic plantations from Darjeeling hill slopes. Esterase I–VI isozymes with higher level of expression were found in specimens from conventional tea plantations than organic populations of H. theivora from Darjeeling hill slopes in isozymes study.     

Association of Genetic Variants with Primary Angle Closure Glaucoma in Two Different Populations

Purpose

A recent large genome-wide association study (GWAS) identified multiple variants associated with primary angle-closure glaucoma (PACG). The present study investigated the role of these variants in two cohorts with PACG recruited from Australia and Nepal.

Method

Patients with PACG and appropriate controls were recruited from eye clinics in Australia (n = 232 cases and n = 288 controls) and Nepal (n = 106 cases and 204 controls). Single nucleotide polymorphisms (SNPs) rs3753841 (COL11A1), rs1015213 (located between PCMTD1 and ST18), rs11024102 (PLEKHA7), and rs3788317 (TXNRD2) were selected and genotyped on the Sequenom. Analyses were conducted using PLINK and METAL.

Results

After adjustment for age and sex, SNP rs3753841 was found to be significantly associated with PACG in the Australian cohort (p = 0.017; OR = 1.34). SNPs rs1015213 (p = 0.014; OR 2.35) and rs11024102 (p = 0.039; OR 1.43) were significantly associated with the disease development in the Nepalese cohort. None of these SNPs survived Bonferroni correction (p = 0.05/4 = 0.013). However, in the combined analysis, of both cohorts, rs3753841 and rs1015213 showed significant association with p-values of 0.009 and 0.004, respectively both surviving Bonferroni correction. SNP rs11024102 showed suggestive association with PACG (p-value 0.035) and no association was found with rs3788317.

Conclusion

The present results support the initial GWAS findings, and confirm the SNP’s contribution to PACG. This is the first study to investigate these loci in both Australian Caucasian and Nepalese populations.     

The genome of the gymnosperm Picea glauca encodes a single Nucleobase Cation Symporter 1 (PgNCS1) that displays a broad yet unique solute specificity profile

Plant Cell, Tissue and Organ Culture (PCTOC) - The Picea glauca genome contains a locus that encodes for a nucleobase cation symporter 1 (PgNCS1). As a gymnosperm, P. glauca belongs to a key...