Trimeresurus stejnegeri snake venom plasminogen activator - Site-directed mutagenesis and molecular modeling

The specific plasminogen activator from Trimeresurus stejnegeri venom (TSV-PA) is a serine proteinase presenting 23% sequence identity with the proteinase domain of tissue type plasminogen activator, and 63% with batroxobin, a fibrinogen clotting enzyme from Bothrops atrox venom that does not activate plasminogen. TSV-PA contains six disulfide bonds and has been successfully overexpressed in Escherichia coli (Zhang, Y., Wisner, A., Xiong, Y. L,, and Bon, C, (1995) J. Biol. Chem. 270, 10246-10255), To identify the functional domains of TSV-PA, we focused on three short peptide fragments of TSV-PA showing important sequence differences with batroxobin and other venom serine proteinases. Molecular modeling shows that these sequences are located in surface loop regions, one of which is next to the catalytic site, When these sequences were replaced in TSV-PA by the equivalent batroxobin residues none generated either fibrinogen-clotting or direct fibrinogenolytic activity, Two of the replacements had little effect in general and are not critical to the specificity of TSV-PA for plasminogen. Nevertheless, the third replacement, produced by the conversion of the sequence DDE 96a-98 to NVI, significantly increased the K-m for some tripeptide chromogenic substrates and resulted in undetectable plasminogen activation, indicating the key role that the sequence plays in substrate recognition by the enzyme.

Molecular dynamics simulation on the complex of the tobramycin and 16S rRNA a site

A 3. 6 ns molecular dynamics simulation was carried out on the complex system of tobramycin and 16S rRNA in order to understand the speciality recognition mechanism between tobramycin and 16S rRNA at the molecular level. The results demonstrate that two l

Secondary structural analysis of the mRNA regions encoding the hemagglutinin cleavage site basic amino acids of the avian influenza virus H5N1 subtype samples

Here we report the codon bias and the mRNA secondary structural features of the hemagglutinin (HA) cleavage site basic amino acid regions of avian influenza virus H5N1 subtypes. We have developed a dynamic extended folding strategy to predict RNA secondar

Site-directed PEGylation of trichosanthin retained its anti-HIV activity with reduced potency in vitro

Trichosanthin (TCS) was the first ribosome inactivating protein found to possess anti-HIV-1 activity. Phase I/II clinical trial of this compound had been done. Antigenicity and short plasma half-life were the major side effects preventing further clinical trial. Modification of TCS is therefore necessary to revive the interest to develop this compound as an anti-HIV agent. Three potential antigenic sites (Ser-7, Lys-173, and Gln-219) were identified by computer modeling. Through site-directed mutagenesis, these three antigenic amino acids were mutated to a cysteine residue resulting in 3 TCS mutants, namely S7C, K173C, and Q219C. These mutants were further coupled to polyethylene glycol with a molecular size of 20 kDa (PEG) via the cysteine residue. This produced another three TCS derivatives, namely PEG(20)k-S7C, PEG(20)k-K173C, and PEG(20)k-Q219C. PEGylation had been widely used recently to decrease immunogenicity by masking the antigenic sites and prolong plasma half-life by expanding the molecular size. The in vitro anti-HIV-1 activity of these mutants and derivatives was tested. Results showed that the anti-HIV-1 activity of S7C, K173C, and Q219C was decreased by about 1.5- to 5.5-fold with slightly lower cytotoxicity. On the other hand, PEGylation produced larger decrease (20- to 30-fold) in anti-HIV activity. Cytotoxicity was, however, weakened only slightly by about 3-fold. The in vitro study showed that the anti-HIV activity of PEGylated TCS was retained with reduced potency. The in vivo activity is expected to have only slightly changed due to other beneficial effects like prolonged half-life. (C) 2004 Elsevier Inc. All rights reserved.

Symmetrical 1-pyrrolidineacetamide showing anti-HIV activity through a new binding site on HIV-1 integrase

Aim: To characterize the functional and pharmacological features of a symmetrical 1-pyrrolidineacetamide, N,N'-(methylene-di-4,1-phenylene) bis-1-pyrrolidineacetamide, as a new anti-HIV compound which could competitively inhibit HIV-1 integrase (IN) bindi

Co-management approach on fisher group: a case study on Ramsar site, Tanguar haor in Bangladesh

In Bangladesh, wetlands are managed through leasing system traditionally from time immemorial. Recently the Government accepted co-management approach for wetland fisheries management and this approach is being practiced in few wetlands for maximize revenue income. A study was carried out to evaluate trend and impact of co-management in Tanguar haor (a Ramsar site wetland) on fisheries resources and livelihood of resident people in the immediate vicinity of the wetland. In Tanguar haor, conflict between leaseholders and the local community was a common phenomenon in the past. Since 2003 the district administration of Sunamganj has been managing the vast wetland resources, however, local people participation was ignored in haor management system. Average monthly fish catch of fishermen increased by 17% after introduction of co-management system and 7 fish species reappeared after introduction of co-management. Average monthly volume of fish catch has increased from 70 kg to 87 kg. A well-defined management structure has been developed for integration of all people of Tanguar haor which would enable them to raise voice jointly and influence policy in their favour.

A carp hatchery: its essential components, site, proper hatchery operation procedures

The paper presents the essential components, ideal site, and proper hatchery operation procedures of a carp hatchery.

Nest site selection by Black-necked Crane Grus nigricollis in the Ruoergai Wetland, China

From 5 May 2003 to early June 2005, nest site selection of Black-necked Cranes Grits nigricollis was studied at the Ruoergai Wetland Nature Reserve (RWNR), an important breeding area for the species in China. Results showed that the crane nests only in we

Economic impacts of globalisation at the district, landing site and sub-sector enterprise levels on Lakes Kyoga, Albert and Victoria

The Globalisation and fish utilisation and marketing study is a collaboration between the Fisheries Resources Research Institute (FIRRI) and the Mike Dillon Associates Limited , with funding from the Department for International Development (DFID) of the Government of the United Kingdom. The study is designed to examine the impact of the development of the export fishery on the fish producers, processors, traders and consumers in the artisanal fishery in Uganda. FIRRI 's role is to collect field data relating to the livelihoods of artisanal fish producers, processors, traders and consumers. in particular data relating to income and revenue flow. The initial focus is on the eccnomic structure of fish landing sites. The purpose of this paper is to review the progress in implementation of the project and present the interim findings for discussion. During the first quarter, namely April to June, 2002, work was carried out on Lakes Kyoga and Albert and a report produced. During the second quarter, July to September, 2002, Lake Victoria was covered. In both phases, the focus has been on the economic structure of fish landings.

Report of the baseline survey undertaken on the Chinese cage site in Napoleon Gulf, Northern Lake Victoria, 12 March 2012

The government of the People's Republic of China through a 2007 agreement with the Government of the Republic of Uganda, has establishment of an Agricultural Technology Demonstration Center (ATDC). The first phase covering the building of aquaculture infrastructure at Kajjansi ARDC is complete and the second operation phase has started in which facilities for cage culture have been set up in the Napoleon gulf, northern Lake Victoria near Jinja. The cage facility is aimed at boosting fish farming within the lake as a diversification to the traditional pond fish culture technology. NaFIRRI scientists as well as Chinese experts undertook a baseline survey in the chosen cage site on 12 March 2012. The survey covered determination of water depth, water transparency, measurement of selected physical-chemical parameters (temperature,dissolved oxygen, conductivity and pH; determination of the nutrient status and study of algae, invertebrate and fish communities at the site. Materials and methodologies used in the survey were based on the Standard Operating Procedures (SOPs) of NaFIRRI. The study area was divided into three study sites. Site 1 (upstream) was at 8.9 metre depth while site 2 (proposed cage site) and site 3 (downstream) were 6 and 4.3 metres deep respectively. Water transparency was lowest at site 1 (1.58 m) and highest at site 3 (1.64 m). Dissolved oxygen at the three sites ranged from 6.0 to 8 mg/I. Water temperature profiles fluctuated within narrow limits between 26.5 and 27.5 DC. Measurements of pH were between 7 (neutral) and 8 (alkaline) while electrical conductivity was between 98 and 101 uS/em. These observed physical-chemical parameters at the study site were considered suitable for cage fish rearing purposes. Nitrite-nitrogen levels varied within narrow limits from 0.043 to 0.0453 mgtl. Similarly, Ammonia-nitrogen varied between 0.015 and 0.0185 mg/1. Soluble reactive phosphorus (SRP) level was highest at site 3 (O.012mgll) compared to that at sites 1 and 2 (0.009mgll). Total suspended solids (TSS) were higher at site 1 (83.3mgll), thereafter decreasing to lower levels at sites 2 (24.8mgtl) and 3 (19.8mgl) respectively. The nutrient level results observed here all fall below the maximum permissible limits by NEMA and therefore the site is recommended for cage culture The algal community was constituted by four major groups: Blue greens,Greens, Cryptophytes, and Diatoms with blue greens as the common and dominant group. High algal biomass (19944961 ugtL) of the dominant blue green algae was observed at site 1 compared site 2 and 3 (58655.2 & 27487. 7 ugtL) respectively. Occurrence of toxicin producing algae: microsytis and cylindrospermopsis in the proposed cage area was considered to be of not much significance as their concentrations were below harmful levels. However, monitoring their presence, biomass and seasonality will be critical in order to follow when and where they occur and at what time of the year for ease of management of the cages

Monthly patterns in physico-chemical conditions and fish catch in the immediate vicinity of the Bujagali hydropower construction site 29th September - 28th October 2007

The specific objectives were to: WATER QUALITY 1. To measure the water physical variables as indicators of environmental conditions in the upstream and downstream transects of Kalange (1) and Buyala (2), respectively, 2. To determine the concentrations of total suspended solids as a major constituent likely to be released into the waters at any time during the construction activities, by comparing the concentrations at the two transects. FISH CATCH 1. To follow up trends in fish catch as construction activity progresses, and to precision of the estimate; 2. To estimate the prevailing fish catch rates, total fish catches and the total value of the fish catch to the local fishers at the two transects.