Studies on inorganic carbon uptake and photosynthesis in blue green alga Nostoc calcicola.

Inorganic carbon uptake and utilization efficiency of a diazotrophic cyanobacterium, Nostoc calcicola, an 'usar' land isolate and its bicarbonate resistant mutant was investigated. The wild type strain showed significantly higher rate uptake of inorganic carbon as compared to mutant strain. The rate of photosynthesis and carbohydrate content of wild type strain was higher than mutant strain at ambient atmosphere, while at higher concentrations of inorganic carbon (100-250 mM NaHCO3) mutant strain showed better response. However the photosynthetic rate and carbohydrate content of the mutant strain was higher than wild type at their respective optimal NaHCO3 concentrations (75 and 250mM, respectively). This is indicative of the mutant strain required higher level of bicarbonate in the medium for optimal activity/growth. It may be concluded that the mutant strain is defective in carbon concentrating mechanism (CCM), and may provide a useful tool in understanding of CCM in these organisms, which in turn has a huge potential to act as global CO, sink.

The etiology of fever in patients presented at KIST Medical College, Teaching Hospital, Lalitpur, Nepal.

In Nepal, febrile illness is one of the most common reasons for seeking medical attention, but there is limited information on the frequency of specific etiology of fever. This is a retrospective study. Patients presented with fever admitted in medicine ward and Intensive care unit of KIST Medical College, Imadol, Lalitpur from January 2010 to January 2012 are included in the study. Of the 898 patients enrolled, 23.5% had infective exacerbation of COPD. Enteric fever, urinary tract infection, acute gastroenteritis, tuberculosis and community acquired pneumonia were the cause of fever in 20%, 13%, 10%, 7.3% and 6% cases of respectively. Findings confirm the heavy burden of infection was a cause of fever requiring hospitalization. This highlights the importance of simple diagnostic tests and cost effective treatment required to manage these patients.

Contrasting pattern of methanotrophs in dry tropical forest soils: effect of soil nitrogen, carbon and moisture.

Population dynamics of methane-oxidizing bacteria (MOB) was measured for 2 consecutive years for four forest and one savanna sites in seasonally dry tropical regions of India. The soils were nutrient-poor and well drained. These sites differed in vegetational cover and physico-chemical features of soils. There were significant differences in MOB population size during the 2 years (mean 0.40 and 0.48 x 10(5) cells g(-1) dry soil), and at different sites (mean 0.38-0.59 x 10(5) cells g(-1) dry soil). The mean population size of MOB was higher (P<0.05) in dry seasons than in the rainy season at all the sites. There was a significant season and site interaction, indicating that the effect of different seasons differed across the sites. There was a positive relation between soil moisture and MOB population size during summer (the driest period) and a negative relation during the rest of the year. The number of MOB was consistently higher for the Kotwa hill base site than rest of the sites having higher soil organic C and total N. The results suggested that in seasonally dry tropical forests the moisture, C and N status of the soil regulates the population size of MOB (methanotrophs) in the long term.

Dynamics of viable nitrifier community, N-mineralization and nitrification in seasonally dry tropical forests and savanna.

The study was conducted in Vindhyan region, to assess the N-mineralization, nitrification and size of viable community of ammonium- and nitrite-oxidizing bacteria as affected by different sites and seasons. Six different ecosystems (four forests and two savannas), which differ in terms of topography, vegetation and moisture status, were selected for the present study. The soils of the study sites differ significantly in its physico-chemical properties. The savanna site had significantly higher pH (7.2), bulk density (1.37 g cm(-3)) and silt content (67.80%) but lower water holding capacity (1.37%), total-C (16,356 microg g(-1) dry soil), N (1090 microg g(-1) dry soil) and P (213 microg g(-1) dry soil) than forest sites. The soil moisture content, N-mineralization, nitrification rates and numbers of ammonium- and nitrite-oxidizing bacteria were highest in the wet season and lowest in dry season, while the size of mineral-N (NH4(+)-N and NO3(-)-N) showed a reverse trend at the sites. The N-mineralization, nitrification and nitrifier population size differ significantly across the site and season. The numbers of free-living cells of ammonium- and nitrite-oxidizing bacteria were significantly related to each other and to N-mineralization, nitrification, soil moisture and mineral-N components. The N-mineralization, nitrification and the viable number of nitrifying cells were consistently higher for forest soils compared to savanna sites. It was concluded that soil microbial process (N-mineralization and nitrification) and nitrifier population size were dependent on site topography, vegetation cover and soil moisture status.

Cyanobacteria in Antarctica: ecology, physiology and cold adaptation.

Cyanobacterial species composition of fresh water and terrestrial ecosystems and chemical environment of water in Schirmacher Oasis in Continental Antarctica was investigated. Over 35 species of cyanobacteria were recorded. Diazotrophic species both heterocystous and unicellular contributed more than half to the count except in lake ecosystem. The species composition varied among the fresh water as well as terrestrial ecosystems. The physico-chemical analyses of water revealed its poor nurient content which might have supported the growth of diazotrophic cyanobacteria in an Antarctic environment. Among the cyanobacteria Oscillatoria, Phormidium and Nostoc commune were the dominant flora in most of the habitats. The physiological characteristics of isolated cyanobacteria strains indicated that N2-fixation, nitrate uptake, nitrate-reduction, ammonium-uptake, GS-transferase activity and photosynthesis was unaffected at low temperature (5 degrees C) which indicated low temperature adaptation for Antarctic cyanobacteria. This phenomenon was not evident in different strains of tropical origin. The temperature optima for N2-fixation for the different Antarctic cyanobacterial strains was in the range of 15-25 degrees C, nearly 10 degrees C lower than their respective reference strains of tropical origin. Similar results were obtained for cyanobacteria-moss association. The low endergonic activation energy exhibited by the above metabolic activities supported the view that cyanobacteria were adapted to Antarctic ecosystem.

An assessment of biopotential of three cyanobacterial isolates from Antarctic for carotenoid production.

Specific growth rates and carotenoid contents of three Antarctic and tropical strains of cyanobacteria viz. Anabaena sp., Phormidium sp. and Nostoc sp. were compared in batch and mass cultures to assess bio-potential of Antarctic strains for cost-effective carotenoid production. Antarctic strains though exhibited slightly lower specific growth rates, but contained higher carotenoid contents (per unit dry wt.), than tropical strains. Modification of normal composition of BG-11 culture medium, by altering nitrogen and carbon sources resulted in 25-38% increase in carotenoid content in both types of strains. Mass-culture in indoor and semi-outdoor bio-reactors resulted in 39-113% higher carotenoid content in Antarctic strains, compared to their respective tropical strains. The observations suggest that Antarctic cyanobacteria may have potential as superior strains for maximizing the yield of carotenoids.

Increased potency of an erythropoietin peptide mimetic through covalent dimerization.

We have synthesized a chemically defined, dimeric form of an erythropoietin mimetic peptide (EMP) that displays 100-fold increased affinity for the erythropoietin receptor (EPOR) and correspondingly elevated potency in cell-based assays and in mice. The dimeric EMP1 was synthesized using a C-terminal lysine residue as a branch point. A beta-alanine residue was coupled to the main-chain (alpha) amino group of the lysine residue in order to provide a pseudosymmetrical scaffold where both the side-chain and main-chain were of approximately equal length. Using an orthogonal protection system, independently disulphide-cylized EMP1 moieties were synthesized upon this scaffold. The proposed mechanism of increased potency of the dimer over the parental compound EMP1 is consistent with the structure of a cocrystal of EMP1 and the extracellular domain of the EPOR in which a noncovalent peptide dimer is seen spanning the cleft between two molecules of the EPOR extracellular domain.

Small peptides as potent mimetics of the protein hormone erythropoietin.

Random phage display peptide libraries and affinity selective methods were used to isolate small peptides that bind to and activate the receptor for the cytokine erythropoietin (EPO). In a panel of in vitro biological assays, the peptides act as full agonists and they can also stimulate erythropoiesis in mice. These agonists are represented by a 14- amino acid disulfide-bonded, cyclic peptide with the minimum consensus sequence YXCXXGPXTWXCXP, where X represents positions allowing occupation by several amino acids. The amino acid sequences of these peptides are not found in the primary sequence of EPO. The signaling pathways activated by these peptides appear to be identical to those induced by the natural ligand. This discovery may form the basis for the design of small molecule mimetics of EPO.

Energetics of cyanophage N-1 multiplication in the diazotrophic cyanobacterium Nostoc muscorum.

Cyanophage N-1 multiplication was investigated during the latent period of the virus, when 14CO2 fixation was inhibited whereas respiratory O2 uptake increased approximately 67% at 4 h after infection. A simultaneous decrease (70%) in the glycogen content of infected cells indicated its catabolic involvement. A chloramphenicol-sensitive rise in glucose-6-phosphate dehydrogenase activity as a result of N-1 infection partly explained the increase in aerobic respiration. The total ATP pool declined to 53% of the control while Ca(2+)-dependent ATPase activity also declined (25%). In contrast, Mg(2+)-dependent ATPase activity increased (80%) in comparison with uninfected cells. Results suggest that oxidative phosphorylation was more crucial in the control of cyanophage N-1 development than photophosphorylation under photoautotrophic growth conditions.

Tyrosine-induced heterocyst division in Nostoc muscorum.

Heterocyst size variation in Nostoc muscorum has been surveyed in the presence and absence of tyrosine. The heterocyst size exhibited two major peaks under both conditions but one of the peaks shifted towards larger size in tyrosine-containing medium. Heterocysts of larger volume exhibited division in the latter medium which was not observed in medium lacking tyrosine. It is suggested that signals for cell division did not decay following differentiation of heterocyst in the presence of tyrosine.

ATP-dependent uptake of nitrate in Nostoc muscorum and inhibition by ammonium ions.

A high rate of nitrate uptake was observed in Nostoc muscorum when cells were grown on elemental nitrogen as compared to that when they were grown on nitrate or ammonium. The uptake of nitrate was light dependent. However, supplementation with ATP (50 microM) stimulated nitrate uptake both in light and darkness. ADP, under similar conditions had no effect. 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), 2-n-heptyl-4-hydroxyquinoline, (HOQNO) and KCN inhibited nitrate uptake in light which could be partially reversed by addition of ATP. Inhibition by carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP), an uncoupler of photophosphorylation, was complete and could not be restored by the addition of ATP. N,N'-dicyclohexylcarbodiimide (DCCD), a specific inhibitor of ATPase, blocked nitrate uptake in the presence or absence of externally added ATP. Although no nitrate uptake was observed under anaerobic conditions in dark, addition of ATP resulted in uptake of nitrate, which was similar in magnitude to that observed under aerobic condition in the light, and was inhibited by DCCD. Ammonium ions inhibited the uptake of nitrate in the absence of ATP but in its presence there was simultaneous uptake of nitrate and ammonium ions. However, uptake of ammonium ions alone was not affected by presence or absence of ATP in the external medium. It was concluded that nitrate ion uptake was energy dependent and may be linked with a proton gradient which can be formed either by photophosphorylation or ATP hydrolysis.