Factors effecting the solubilisation of stearoyl-coA desaturase of hen liver microsomes.

1. The lipid requirement for maximum desaturase activity was investigated using acetone/water mixtures. It was shown that for maximum stearoyl-CoA desaturase activity of hen liver microsomes neither the total neutral lipid fraction nor 44% of the phospholipid fraction were required. 2. The effect of sodium deoxycholate, Triton X-100, Nonidet P-40 and Bio-solv on the enzyme activity indicated that the neutral detergents had a milder effect than the ionic detergent but both classes could cause considerable irreversible loss of activity. 3. The treatment of the microsomes with 2.5% (v/v) water in acetone greatly improved the effective solubilising power of Triton X-100. The yield of desaturase in the 100 000 X g supernatant obtained by treating the microsomal fraction in this way was strongly dependent upon protein concentration. Maximum solubilisation was achieved with25 mg protein per ml 1% (w/v) Triton X-100 in 0.1 M potassium phosphate buffer pH 7.4. 4. A comparison of the properties of the solubilised and membrane-bound enzyme was made by an investigation of: (i) the temperature and pH optimum, (ii) activation energy and (iii) the effect of inhibitors on the enzyme activity.     

A novel harmony search-K means hybrid algorithm for clustering gene expression data

Recent progress in bioinformatics research has led to the accumulation of huge quantities of biological data at various data sources. The DNA microarray technology makes it possible to simultaneously analyze large number of genes across different samples. Clustering of microarray data can reveal the hidden gene expression patterns from large quantities of expression data that in turn offers tremendous possibilities in functional genomics, comparative genomics, disease diagnosis and drug development. The k- ¬means clustering algorithm is widely used for many practical applications. But the original k-¬means algorithm has several drawbacks. It is computationally expensive and generates locally optimal solutions based on the random choice of the initial centroids. Several methods have been proposed in the literature for improving the performance of the k-¬means algorithm. A meta-heuristic optimization algorithm named harmony search helps find out near-global optimal solutions by searching the entire solution space. Low clustering accuracy of the existing algorithms limits their use in many crucial applications of life sciences. In this paper we propose a novel Harmony Search-K means Hybrid (HSKH) algorithm for clustering the gene expression data. Experimental results show that the proposed algorithm produces clusters with better accuracy in comparison with the existing algorithms.     

Enhancement of seed phytosterol levels by expression of an N-terminal truncated Hevea brasiliensis (rubber tree) 3-hydroxy-3-methylglutaryl-CoA reductase

Dietary intake of phytosterols (plant sterols) has been shown to be effective in reducing blood cholesterol levels, thereby reducing the risk of cardiovascular disease. Phytosterols are most commonly sourced from vegetable oils, where they are present as minor components. We report here the generation of transgenic tobacco seeds substantially enhanced in phytosterol content by the expression of a modified form of one of the key sterol biosynthetic enzymes, 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR). The constitutive expression of an N-terminal truncated Hevea brasiliensis HMGR (t-HMGR), lacking the membrane binding domain, enhanced seed HMGR activities by 11-fold, leading to increases in total seed sterol of 2.4-fold. Seed-specific expression of t-HMGR enhanced total seed sterol levels by 3.2-fold, to 1.36% dry weight or 3.25% of oil. 4-desmethylsterols were increased by 2.2-fold, whilst certain sterol biosynthetic intermediates, in particular cycloartenol and 24-ethylidene lophenol, also accumulated. The additional sterol in seed tissue was present in the form of fatty acid esters. Constitutive expression of t-HMGR increased leaf phytosterol sterol levels by 10-fold, representing 1.8% dry weight, and the sterol was sequestered, in acyl ester form, as cytoplasmic 'oil droplets'. These studies establish HMGR as a key enzyme controlling overall flux into the sterol biosynthesis pathway in seed tissue, but the accumulation of certain intermediates suggests additional slow steps in the pathway. The expression of an N-truncated HMGR activity has generated novel phytosterol-enriched raw materials that may provide the basis of new sourcing opportunities for this important class of cholesterol-lowering actives.     

Application of magnetic techniques in the field of drug discovery and biomedicine

Magnetic separation technology, using magnetic particles, is quick and easy method for sensitive and reliable capture of specific proteins, genetic material and other biomolecules. The technique offers an advantage in terms of subjecting the analyte to very little mechanical stress compared to other methods. Secondly, these methods are non-laborious, cheap and often highly scalable. Moreover, techniques employing magnetism are more amenable to automation and miniaturization. Now that the human genome is sequenced and about 30,000 genes are annotated, the next step is to identify the function of these individual genes, carrying out genotyping studies for allelic variation and SNP analysis, ultimately leading to identification of novel drug targets. In this post-genomic era, technologies based on magnetic separation are becoming an integral part of todays biology laboratory. This article briefly reviews the selected applications of magnetic separation techniques in the field of biotechnology, biomedicine and drug discovery.     

Phenotypic and Genetic Diversity of Aeromonas Species Isolated from Fresh Water Lakes in Malaysia

Gram-negative bacilli of the genus Aeromonas are primarily inhabitants of the aquatic environment. Humans acquire this organism from a wide range of food and water sources as well as during aquatic recreational activities. In the present study, the diversity and distribution of Aeromonas species from freshwater lakes in Malaysia was investigated using glycerophospholipid-cholesterol acyltransferase (GCAT) and RNA polymerase sigma-factor (rpoD) genes for speciation. A total of 122 possible Aeromonas strains were isolated and confirmed to genus level using the API20E system. The clonality of the isolates was investigated using ERIC-PCR and 20 duplicate isolates were excluded from the study. The specific GCAT-PCR identified all isolates as belonging to the genus Aeromonas, in agreement with the biochemical identification. A phylogenetic tree was constructed using the rpoD gene sequence and all 102 isolates were identified as: A. veronii 43%, A. jandaei 37%, A. hydrophila 6%, A. caviae 4%, A. salmonicida 2%, A. media 2%, A. allosaccharophila 1%, A. dhakensis 1% and Aeromonas spp. 4%. Twelve virulence genes were present in the following proportions—exu 96%, ser 93%, aer 87%, fla 83%, enolase 70%, ela 62%, act 54%, aexT 33%, lip 16%, dam 16%, alt 8% and ast 4%, and at least 2 of these genes were present in all 102 strains. The ascV, aexU and hlyA genes were not detected among the isolates. A. hydrophila was the main species containing virulence genes alt and ast either present alone or in combination. It is possible that different mechanisms may be used by each genospecies to demonstrate virulence. In summary, with the use of GCAT and rpoD genes, unambiguous identification of Aeromonas species is possible and provides valuable data on the phylogenetic diversity of the organism.     

Historical trends in frog populations in New Zealand based on public perceptions

Surveys were distributed to New Zealand land users in 1998 and 2008 to acquire information about New Zealand frogs with the aim of compiling and mapping their distribution and inferred population trends without costly and time-consuming field surveys. The overall frog population trend was reported as declining, with possible causes reported as an increase in agriculture, an increase in the distribution of predatory fish and disease. The resultant maps could be used for four main purposes: 1) to identify regions where Litoria populations are known to occur, which can be eliminated when considering suitable regions for translocation of Leiopelma; 2) to identify growing or stable populations of Litoria species, which may assist future disease surveys, population monitoring and to identify sources of genetic material that may serve as an Ark for declining Australian populations; 3) to highlight populations that are in decline to enable effective targeting of detailed disease studies; and 4) to approximate the stability of amphibian populations in the absence of more accurate, but costly, scientific monitoring.     

BAER Soil Burn Severity Maps Do Not Measure Fire Effects to Vegetation: A Comment on Odion and Hanson (2006)

Abstract We comment on a recent Ecosystem paper by Odion and Hanson (Ecosystems 9:1177–1189, 2006), in which the authors claim that high severity fire is rare in the Sierra Nevada under current conditions. Odion and Hanson’s results are predicated on BAER soil burn severity maps, which are based primarily on fire effects to soil, not vegetation. Odion and Hanson, and we fear others as well, are misinformed as to the nature of the BAER severity mapping process, and proper applications of BAER soil burn severity maps. By comparing the BAER soil burn severity maps to a true vegetation burn severity measure (RdNBR) calibrated by field data, we show that the area in the high soil burn severity class for the three fires analyzed by Odion and Hanson is substantially less than the area of stand-replacing fire, and that BAER maps—especially hand-derived maps such as those from two of the three fires—also greatly underestimate the heterogeneity in vegetation burn severity on burned landscapes. We also show that, contrary to Odion and Hanson’s claims, Fire Return Interval Departure (FRID) is strongly correlated with fire severity in conifer stands within the perimeter of the McNally Fire.     

NOTES ON THE BIOLOGY OF THE MAURITIUS BLACK BULBUL HYPSIPETES OLIVACEUS

Safford, R.J. 1996. Notes on the biology of the Mauritius Black Bulbul Hypsipetes olivaceus. Ostrich 67: 151–154.

The Mauritius Black Bulbul Hypsipetes olivaceus is a threatened passerine endemic to Mauritius. Direct observations of food items indicate that it is an arboreal omnivore. Animal prey consists mostly of insects and native day-geckoes (Phelsuma spp.). A wide variety of native and exotic fruits is also taken. The species is apparently monogamous, with an egg-laying season extending from at least November to February, usual clutch and brood sizes of two to three, ability rapidly to lay repeat clutches in a new nest, and post-breeding complete moult. In these respects, its annual cycle resembles that of the other native Mauritian passerines. It occurs at very low population density compared to its Indian Ocean congeners; possible reasons for this are given.     

The lemurs of marojejy strict nature reserve,Madagascar: A status overview with notes on ecology and threats

From mid-August to late October 1988, we surveyed Marojejy Strict Nature Reserve, in the northern part of Madagascar’s rain forest. Although widely believed to be a keystone site for lemur conservation, only incidental information concerning Marojejy’s primate communities has hitherto been published. The reserve extends from 75 to 2133 m in altitude, and its 60,150 ha comprise an almost-intact series of altitudinal forest zones, now fundamentally isolated from formerly contiguous surrounding forest. We visited all forest zones and all altitudes of the reserve. We observed a total of nine species including the little-known diademed sifaka (Propithecus diadema candidus),for which Marojejy is the most important site. Aye-aye (Daubentonia madagascarensis)is also present. Brown lemur (Eulemur fulvus albifrons) isthe most frequently observed species. Little is known about this subspecies in the wild. In the interior, many lemurs were extremely inquisitive of human presence, showing how little hunted they are. This contrasted with their lower numbers and evasive behavior in the more accessible regions of the reserve. The major threats to the lemurs of Marojejy are the ceaseless piecemeal clearance of the reserve’s forest, which is proceeding inward from the boundary, and direct trapping.