The status of Lake Lanao endemic cyprinids (Puntius species) and their conservation

The endemic cyprinid species flock of Lake Lanao, Philippines is a classic example of explosive evolution. Based on historical surveys, most of the 18 endemic cyprinid species in Lake Lanao are now believed to be extinct. We compared results of landing and market surveys conducted in 2008 to information from historical surveys. Field sampling and fishermen interviews provided additional information on the status of the endemic and other fish species in Lake Lanao. Puntius lindog and Puntius tumba were the only two endemic species recorded in 2008, accounting for 0.01 % and 0.04 % respectively of the total weight harvested. Our discovery that at least two endemic cyprinids remain in Lake Lanao is surprising and significant as the lake is increasingly imperiled by human disturbances, exploitation and introduced species. Protecting and conserving the remaining endemic species are important management priorities.     

Impact of protein diversification on morphometric behavior of Andrographis paniculata Nees

Proteins are important biochemical parameters in genetic diversity and controlling morphological characteristics in plants. In this study, the proteomic and morphometric data of an important medicinal herb “Aci Pa?a” (Andrographis paniculata) were combined together to illustrate their impacts on genetic variation of the plant’s population and to realize the connection between protein patterns and phenotypic behavior of the species. We used three protein extraction buffers including Tris, potassium phosphate, and sodium citrate. The Tris buffer was significantly different (p ≤ 0.01) than other two in terms of the quality and quantity of protein bands by producing 15 types of proteins ranged from 13 to 105 kDa of which two of them were polymorphic. Consequently, a total of 12 accessions of A. paniculata were subjected to morpho-proteomic analyses. The unweighted pair group method with arithmetic average cluster analysis of the accessions based on the protein data and morphological characteristics generated three and four clusters, respectively, at a Euclidean distance of 2.53 for the morphological traits. Moreover, seed proteins analysis revealed that the two polymorphic protein bands sized 20.5 (protein “b”) and 30 kDa (protein “a”) effectively diversified the morphological characteristics and phylogenetic relationships among the 12 accessions of A. paniculata. Interestingly, the protein “b” acted as an activator agent for the number of branches, leaves and total dry weight, while the protein “a” performed a suppressive role for the same traits. Additionally, the two high-weighted faint bands “c” (75 kDa) and “d” (100 kDa) with a very low expression in accession 11228 proved their suppressive role along with the “a” band, while these bands were strongly expressed in the rest of the accessions. These findings suggest that these four proteins should be sequenced and perfectly established for further proteomic analyses. Ultimately, the mentioned proteins can be developed for any prospective breeding program or gene identification.     

Comparative proteomic analysis of different developmental stages of the edible mushroom Termitomyces heimii

Background

Termitomyces heimii is a basidiomycete fungus that has a symbiotic relationship with termites, and it is an edible mushroom with a unique flavour and texture. T. heimii is also one of the most difficult mushrooms to cultivate throughout the world. Little is known about the growth and development of these mushrooms, and the available information is insufficient or poor. The purpose of this study was to provide a base of knowledge regarding the biological processes involved in the development of T. heimii. The proteomic method of 2 dimensional difference gel electrophoresis 2D-DIGE was used to determine and examine the protein profiles of each developmental stage (mycelium, primordium and fruiting body). Total proteins were extracted by TCA-acetone precipitation.

Results

A total of 271 protein spots were detected by electrophoresis covering pH 3–10 and 10–250 kDa. Selected protein spots were subjected to mass spectrometric analyses with matrix-assisted laser desorption/ionisation (MALDI TOF/TOF). Nineteen protein spots were identified based on peptide mass fingerprinting by matching peptide fragments to the NCBI non-redundant database using MASCOT software. The 19 protein spots were categorised into four major groups through KEGG pathway analysis, as follows: carbohydrate metabolism, energy metabolism, amino acid metabolism and response to environmental stress.

Conclusions

The results from our study show that there is a clear correlation between the changes in protein expression that occur during different developmental stages. Enzymes related to cell wall synthesis were most highly expressed during fruiting body formation compared to the mycelium and primordial stages. Moreover, enzymes involved in cell wall component degradation were up-regulated in the earlier stages of mushroom development.     

A Modified Lumry–Eyring Analysis for the Determination of the Predominant Mechanism Underlying the Diminution of Protein Aggregation by Glycerol

Aggregation of aspartate-β-semialdehyde dehydrogenase (ASD) was analyzed by applying modified Lumry–Eyring with nucleated polymerization (LENP) model. Intrinsic nucleation time scales were determined. In absence of glycerol, ASD undergoes concentration and time-dependent polymerization into low-molecular weight soluble aggregates and thereafter condensation into insoluble aggregates. In the presence of increasing solvent glycerol concentration, the aggregation becomes more and more nucleation dominated, with slower polymerization to low-molecular weights soluble aggregates, without any condensation into insoluble aggregates. Effective nucleus size as well as the number of monomers in each irreversible growth event were sensitive to the changes in solvent glycerol concentration. Glycerol-directed diminution of aggregation appears to be largely due to the inhibition of rearrangement (decreased nucleation rearrangement rate coefficient, K _r,x ) because of compaction induced due to preferential hydration, thus, preventing the soluble aggregates from locking into irreversible soluble nuclei. Appreciably decreased K _r,x (as compared to nucleation dissociation constant, K _d,x ), appears to be responsible for increased nucleus size at higher solvent glycerol concentration. This study explains how modified LENP model can be applied to determine the predominant mechanism responsible for the diminution of aggregation by polyhydric alcohols (glycerol).     

Identification and validation of T-cell epitopes in outer membrane protein (OMP) of Salmonella typhi

This study aims to design epitope-based peptides for the utility of vaccine development by targeting outer membrane protein F (Omp F), because two available licensed vaccines, live oral Ty21a and injectable polysaccharide, are 50% to 80% protective with a higher rate of side effects. Conventional vaccines take longer time for development and have less differentiation power between vaccinated and infected cells. On the other hand, Peptide-based vaccines present few advantages over other vaccines, such as stability of peptide, ease to manufacture, better storage, avoidance of infectious agents during manufacture, and different molecules can be linked with peptides to enhance their immunogenicity. Omp F is highly conserved and facilitates attachment and fusion of Salmonella typhi with host cells. Using various databases and tools, immune parameters of conserved sequences from Omp F of different isolates of Salmonella typhi were tested to predict probable epitopes. Binding analysis of the peptides with MHC molecules, epitopes conservancy, population coverage, and linear B cell epitope prediction were analyzed. Among all those predicted peptides, ESYTDMAPY epitope interacted with six MHC alleles and it shows highest amount of interaction compared to others. The cumulative population coverage for these epitopes as vaccine candidates was approximately 70%. Structural analysis suggested that epitope ESYTDMAPY fitted well into the epitope-binding groove of HLA-C*12:03, as this HLA molecule was common which interact with each and every predicted epitopes. So, this potential epitope may be linked with other molecules to enhance its immunogenicity and used for vaccine development.     

Structure based molecular inhibition of Caspase-8 for treatment of multi-neurodegenerative diseases using known natural compounds

Neurodegenerative disorders are often associated with excessive neuronal apoptosis. It is well known that apoptosis is regulated by some intracellular proteases, such as, Caspases (cysteine-dependent, aspartate-specific proteases). In fact, Caspase-8 which is an initiator caspase, has been identified as a key mediator of neuronal apoptosis. In addition, Caspase-8 is found to be coupled with the regulation of various neurodegenerative disorders including Alzheimer׳s disease (AD), Parkinson׳s disease (PD), Huntington׳s Diseases (HD) and Dentatorubral Pallidoluysian Atrophy (DRPLA). Caspase-8 inhibition may provide an effective means of treatment for multiple neurodegenerative disorders. Therefore, the present study describes the molecular interaction of some selected natural compounds with known anti neurodegenerative properties with Caspase-8. Docking between Caspase-8 and each of these compounds (separately) was performed using ‘Autodock4.2’. Out of all the selected compounds, rosmarinic acid and curcumin proved to be the most potent inhibitors of Caspase-8 with binding energy (ΔG) of -7.10 Kcal/mol and -7.08 Kcal/mol, respectively. However, further in vitro and in vivo studies are needed to validate the anti-neurodegenerative potential of these compounds.     

A Subset of Circulating Blood Mycobacteria-Specific CD4 T Cells Can Predict the Time to Mycobacterium tuberculosis Sputum Culture Conversion

We investigated 18 HIV-negative patients with MDR-TB for M. tuberculosis (Mtb)- and PPD-specific CD4 T cell responses and followed them over 6 months of drug therapy. Twelve of these patients were sputum culture (SC) positive and six patients were SC negative upon enrollment. Our aim was to identify a subset of mycobacteria-specific CD4 T cells that would predict time to culture conversion. The total frequency of mycobacteria-specific CD4 T cells at baseline could not distinguish patients showing positive or negative SC. However, a greater proportion of late-differentiated (LD) Mtb- and PPD-specific memory CD4 T cells was found in SC positive patients than in those who were SC negative (p = 0.004 and p = 0.0012, respectively). Similarly, a higher co-expression of HLA-DR⁺Ki67⁺ on Mtb- and PPD-specific CD4 T cells could also discriminate between sputum SC positive versus SC negative (p = 0.004 and p = 0.001, respectively). Receiver operating characteristic (ROC) analysis revealed that baseline levels of Ki67⁺HLA-DR⁺ Mtb- and PPD-specific CD4 T cells were predictive of the time to sputum culture conversion, with area-under-the-curve of 0.8 (p = 0.027). Upon treatment, there was a significant decline of these Ki67⁺HLA-DR⁺ T cell populations in the first 2 months, with a progressive increase in mycobacteria-specific polyfunctional IFNγ⁺IL2⁺TNFα⁺ CD4 T cells over 6 months. Thus, a subset of activated and proliferating mycobacterial-specific CD4 T cells (Ki67⁺HLA-DR⁺) may provide a valuable marker in peripheral blood that predicts time to sputum culture conversion in TB patients at the start of treatment.     

Effect of Geometric Parameters on the Performance of P-Type Junctionless Lateral Gate Transistors

This paper examines the impact of two important geometrical parameters, namely the thickness and source/drain extensions on the performance of low doped p-type double lateral gate junctionless transistors (DGJLTs). The three dimensional Technology Computer-Aided Design simulation is implemented to calculate the characteristics of the devices with different thickness and source/drain extension and based on that, the parameters such as threshold voltage, transconductance and resistance in saturation region are analyzed. In addition, simulation results provide a physical explanation for the variation of device characteristics given by the variation of geometric parameters, mainly based on investigation of the electric field components and the carries density variation. It is shown that, the variation of the carrier density is the main factor which affects the characteristics of the device when the device''s thickness is varied. However, the electric field is mainly responsible for variation of the characteristics when the source/drain extension is changed.     

Determinants of Public Attitudes to Genetically Modified Salmon

The objective of this paper is to assess the attitude of Malaysian stakeholders to genetically modified (GM) salmon and to identify the factors that influence their acceptance of GM salmon using a structural equation model. A survey was carried out on 434 representatives from various stakeholder groups in the Klang Valley region of Malaysia. Public attitude towards GM salmon was measured using self-developed questionnaires with seven-point Likert scales. The findings of this study have confirmed that public attitudes towards GM salmon is a complex issue and should be seen as a multi-faceted process. The most important direct predictors for the encouragement of GM salmon are the specific application-linked perceptions about religious acceptability of GM salmon followed by perceived risks and benefits, familiarity, and general promise of modern biotechnology. Encouragement of GM salmon also involves the interplay among other factors such as general concerns of biotechnology, threatening the natural order of things, the need for labeling, the need for patenting, confidence in regulation, and societal values. The research findings can serve as a database that will be useful for understanding the social construct of public attitude towards GM foods in a developing country.     

Modeling of crude oil biodegradation using two phase partitioning bioreactor

In this work, crude oil biodegradation has been optimized in a solid‐liquid two phase partitioning bioreactor (TPPB) by applying a response surface methodology based d ‐optimal design. Three key factors including phase ratio, substrate concentration in solid organic phase, and sodium chloride concentration in aqueous phase were taken as independent variables, while the efficiency of the biodegradation of absorbed crude oil on polymer beads was considered to be the dependent variable. Commercial thermoplastic polyurethane (Desmopan®) was used as the solid phase in the TPPB. The designed experiments were carried out batch wise using a mixed acclimatized bacterial consortium. Optimum combinations of key factors with a statistically significant cubic model were used to maximize biodegradation in the TPPB. The validity of the model was successfully verified by the good agreement between the model‐predicted and experimental results. When applying the optimum parameters, gas chromatography‐mass spectrometry showed a significant reduction in n‐alkanes and low molecular weight polycyclic aromatic hydrocarbons. This consequently highlights the practical applicability of TPPB in crude oil biodegradation. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:797–805, 2014