Polyamine Metabolism in the Roots of Phaseolus vulgaris. Interaction of the Inhibitors of Polyamine Biosynthesis with Putrescine in Growth and Polyamine Biosynthesis

The effects of the inhibitors of polyamine biosynthesis, canavanineand -methyl ornithine on growth, the activities of argininedecarboxylase (EC 4.1.1.19 [EC] ) and ornithine decarboxylase (EC4.1.1.17 [EC] ) and on polyamine content were examined in two differentgrowth regions of Phaseolus vulgaris L. cv. Taylor's Horticulturalroots. Separately, in the same manner, in the same bean rootsystem exogenous putrescine effect and the interaction of canavaninewith putrescine were determined. The arginine and ornithine decarboxylase activities found inroot apex were high where cell division activity was highest.Polyamine (putrescine and spermine) content did not correlatewith these activities, but polyamine level was high in the rootbase where cell elongation is the main process. The arginineanalogue, canavanine, inhibited arginine decayboxylase activityand polymine liters. Putrescine partially reversed the canavanineinhibition of root growth as well as arginine decarboxylaseactivity and polyamine content. Similarly -methyl ornithineslightly inhibited the root length and ornithine decarboxylaseactivity in the root apex. Besides, exogenous putrescine didnot effect significantly the endogenous polyamine titers. Theseresults reinforce the growing connection between polyaminesand the rates of cell devision in the roots of bean plants.Separately, arginine decarboxylase is the main enzyme in thebean roots. (Received November 10, 1986; Accepted March 3, 1987)     

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.     

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.     

Synthesis and carbonic anhydrase inhibitory properties of sulfamides structurally related to dopamine

A series of novel sulfamides incorporating the dopamine scaffold were synthesized. Reaction of amines and tert-butyl-alcohol/benzyl alcohol in the presence of chlorosulfonyl isocyanate (CSI) afforded sulfamoyl carbamates, which were converted to the title compounds by treatment with trifluoroacetic acid or by palladium-catalyzed hydrogenolysis. Inhibition of six α-carbonic anhydrases (CAs, EC 4.2.1.1), that is, CA I, CA II, CA VA, CA IX, CA XII and CA XIV, and two β-CAs from Candida glabrata (CgCA) and Mycobacterium tuberculosis (Rv3588) with these sulfamides was investigated. All CA isozymes were inhibited in the low micromolar to nanomolar range by the dopamine sulfamide analogues. K_is were in the range of 0.061–1.822 μM for CA I, 1.47–2.94 nM for CA II, 2.25–3.34 μM for CA VA, 0.041–0.37 μM for CA IX, 0.021–1.52 μM for CA XII, 0.007–0.219 μM for CA XIV, 0.35–5.31 μM for CgCA and 0.465–4.29 μM for Rv3588. The synthesized sulfamides may lead to inhibitors targeting medicinally relevant CA isoforms with potential applications as antiepileptic, antiobesity antitumor agents or anti-infective.     

Dissecting protein‐protein interactions in proteasome assembly: Implication to its self‐assembly

The 26S proteasome is a multi‐catalytic ATP‐dependent protease complex that recognizes and cleaves damaged or misfolded proteins to maintain cellular homeostasis. The 26S subunit consists of 20S core and 19S regulatory particles. 20S core particle consists of a stack of heptameric alpha and beta subunits. To elucidate the structure‐function relationship, we have dissected protein‐protein interfaces of 20S core particle and analyzed structural and physiochemical properties of intra‐alpha, intra‐beta, inter‐beta, and alpha‐beta interfaces. Furthermore, we have studied the evolutionary conservation of 20S core particle. We find the size of intra‐alpha interfaces is significantly larger and is more hydrophobic compared with other interfaces. Inter‐beta interfaces are well packed, more polar, and have higher salt‐bridge density than other interfaces. In proteasome assembly, residues in beta subunits are better conserved than alpha subunits, while multi‐interface residues are the most conserved. Among all the residues at the interfaces of both alpha and beta subunits, Gly is highly conserved. The largest size of intra‐alpha interfaces complies with the hypothesis that large interfaces form first during the 20S assembly. The tight packing of inter‐beta interfaces makes the core particle impenetrable from outer wall of the cylinder. Comparing the three domains, eukaryotes have large and well‐packed interfaces followed by archaea and bacteria. Our findings provide a structural basis of assembly of 20S core particle in all the three domains of life.     

Isolation and characterization of the tobramycin biosynthetic gene cluster from Streptomyces tenebrarius

The biosynthetic gene cluster for tobramycin, a 2-deoxystreptamine-containing aminoglycoside antibiotic, was isolated from Streptomyces tenebrarius ATCC 17920. A genomic library of S. tenebrarius was constructed, and a cosmid, pST51, was isolated by the probes based on the core regions of 2-deoxy-scyllo-inosose (DOI) synthase, and L-glutamine:DOI aminotransferase and L-glutamine:scyllo-inosose aminotransferase. Sequencing of 33.9 kb revealed 24 open reading frames (ORFs) including putative tobramycin biosynthetic genes. We demonstrated that one of these ORFs, tbmA, encodes DOI synthase by in vitro enzyme assay of the purified protein. The catalytic residues of TbmA and dehydroquinate synthase were studied by homology modeling. The gene cluster found is likely to be involved in the biosynthesis of tobramycin.