Isolation and characterization of microsatellite loci in the Pacific pleasure oyster,Crassostrea corteziensis,and their cross‐species amplification in four other oyster species

Microsatellite loci were isolated in Crassostrea corteziensis using (GT)_n, (CT)_n and (CTGT)_n‐enriched genomic libraries. Within each of 45 sequenced clones, an average of three microsatellite regions (156 total) were observed. Thirty‐three primers were designed, from which 11 microsatellite loci amplified. Ten of those were polymorphic, with a range of two to 30 alleles. Three loci were not in Hardy–Weinberg equilibrium, and linkage disequilibrium was found for six pairs of loci. These microsatellite loci will be further tested for segregation distortions and null alleles to establish a set for population genetic studies of the species in the Northwest coasts of Mexico, and for optimization of aquaculture development. Seven of the microsatellite loci cross‐amplified in Crassostrea palmula, a sympatric species, and will be useful in further genetic studies.     

Seed micromorphology in the genus Neotinea Rchb.f. (Orchidaceae, Orchidinae)

The ultrastructure of testa seed in the genus Neotinea (Orchidaceae, Orchidinae) was examined for the first time. The morphology of the seed and of the anticlinal and periclinal walls was analysed by scanning electron microscopy. Quantitative data concerning the length and width of the seed and embryo, seed and embryo volume, free air space, and number of cells along the longitudinal axis are presented. In all species, the seeds are fusiform in shape with transverse ridges on the inner periclinal walls. This ornamentation pattern is characteristic for the genus Neotinea . It is a good diagnostic value supporting the monophyly of this genus, which has recently been proposed by several authors.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 153 , 133–140.     

Synthesis and biological evaluation of novel cYY analogues targeting Mycobacterium tuberculosis CYP121A1

The rise in multidrug resistant (MDR) cases of tuberculosis (TB) has led to the need for the development of TB drugs with different mechanisms of action. The genome sequence of Mycobacterium tuberculosis (Mtb) revealed twenty different genes coding for cytochrome P450s. CYP121A1 catalyzes a CC crosslinking reaction of dicyclotyrosine (cYY) producing mycocyclosin and current research suggests that either mycocyclosin is essential or the overproduction of cYY is toxic to Mtb. A series of 1,4-dibenzyl-2-imidazol-1-yl-methylpiperazine derivatives were designed and synthesised as cYY mimics. The derivatives substituted in the 4-position of the phenyl rings with halides or alkyl group showed promising antimycobacterial activity (MIC 6.25?μg/mL), with the more lipophilic branched alkyl derivatives displaying optimal binding affinity with CYP121A1 (ⁱPr K_D?=?1.6?μM; ^tBu K_D?=?1.2?μM). Computational studies revealed two possible binding modes within the CYP121A1 active site both of which would effectively block cYY from binding.     

Homology model of 1alpha,25-dihydroxyvitamin D3 24-hydroxylase cytochrome P450 24A1 (CYP24A1): active site architecture and ligand binding

Homology models of cytochrome P450 24A1 (CYP24A1) were constructed using three human P450 structures, CYP2C8, CYP2C9 and CYP3A4 as templates for the model building. Using molecular operating environment (MOE) software the lowest energy CYP24A1 model was then assessed for stereochemical quality and side chain environment. Further active site optimisation of the CYP24A1 model built using the CYP3A4 template was performed by molecular dynamics to generate a final CYP24A1 model. The natural substrate, 1,25-dihydroxyvitamin D(3) (calcitriol) and the CYP24 inhibitor (R)-N-(2-(1H-imidazol-1-yl)-2-phenylethyl)-4'-chlorobiphenyl-4-carboxamide ((R)-VID-400) were docked into the model allowing further validation of the active site architecture. Using the docking studies structurally and functionally important residues were identified with subsequent characterisation of secondary structure.     

Agave for tequila and biofuels: an economic assessment and potential opportunities

This paper explores the economic viability of producing biofuels from Agave in Mexico and the potential for it to complement the production of tequila or mescal. We focus on Agave varieties currently being used by the tequila industry to produce two beverages, tequila and mescal, and explore the potential for biofuel production from these plants. Without competing directly with beverage production, we discuss the economic costs and benefits of converting Agave by‐products to liquid fuel as an additional value‐added product and expanding cultivation of Agave on available land. We find that the feedstock cost for biofuel from the Agave piña alone could be more than US$3 L^?1 on average. This is considerably higher than the feedstock costs of corn ethanol and sugarcane ethanol. However, there may be potential to reduce these costs with higher conversion efficiencies or by using sugar present in other parts of the plant. The costs of cellulosic biofuels using the biomass from the entire plant could be lower depending on the conversion efficiency of biomass to fuel and the additional costs of harvesting, collecting and transporting that biomass.     

Exopolysaccharide-mediated silver nanoparticles produced by <Emphasis Type="Italic">Lactobacillus brevis</Emphasis> NM101-1 as antibiotic adjuvant

A green, simple and effective approach was performed to synthesize potent silver nanoparticles using bacterial exopolysaccharide as both a reducing and stabilizing agent. The formation of nanoparticles was first screened by measuring the surface plasmon resonance peak around 400 nm using UV-vis spectroscopy. The morphology of the synthesized AgNPs was determined using TEM, which indicated that the AgNPs were spherical in shape and with an average size of 11–25 nm. The presence of elemental silver of the AgNPs was confirmed by EDX analysis. The possible functional groups of EPS responsible for the reduction and stabilization of AgNPs were evaluated using FTIR. The EPS reduced AgNPs showed excellent antibacterial, and antibiofilm activities against various human pathogenic bacteria. In addition, the efficiency of AgNPs with various broad-spectrum antibiotics against the tested strains was evaluated. It is evident that, the antibacterial and antibiofilm activities of the selected antibiotics were increased in the presence of AgNPs. The increase in activity was more pronounced for gram-negative bacteria Pseudomonas aeruginosa and E. coli. Interestingly, the combination of antibiotics with AgNPs has significantly increased the membrane protein leakage and ROS generation than antibiotics or AgNPs alone. This work supports that AgNPs can be used to enhance the activity of existing antibiotics against gram-negative and gram-positive bacteria for the treatment of infectious diseases.     

The role of ethanol in preventing biofilm formation of Penicillium purpurogenum

The use of fungi in biotechnology requires that no cell loss takes place; a maximal level of cell–nutrient interaction is required to achieve efficient performance. The occurrence of high cell densities or loss of biomass through cell–surface interaction prevents the desired result. The main purpose of adding ethanol was to manipulate the cell–cell and cell–surface adhesion through manipulating cell surface properties. Scanning electron microscopy indicated that the type of surface and its treatment with ethanol controls the adhesion and biofilm formation of Penicillium purpurogenum. Gamma irradiation slightly affected the wettability of polystyrene strips at 0.5 and 1 kGy, thus slightly decreasing the adhesion, but was not as effective as using ethanol to control the adhesion. The presence of ethanol in the media caused a decrease in surface-bound proteins from 0.348 to 0.133 mg/ml, while surface exopolysaccharides showed a minimal decrease. Ethanol induced oxidative stress which reached its peak when 2.5 % v/v ethanol was added to the media; this was represented by both intracellular and extracellular catalase and lipid peroxidation. On the other hand, fungal biomass and pigment showed a decrease as the ethanol concentrations increased. Therefore, ethanol could be employed to control the surface properties of a fungus, and to inhibit biofilm formation to obtain a high surface area for the fungus to be employed in any biotechnological process.     

Conversion of oil waste to valuable fatty acids using Oleaginous yeast

Oil waste poses a highly dangerous threat to the environment, mainly because it is considered a high energy demanding degradation process. Oleaginous yeast utilizing oil waste to produce microbial fatty acids is considered an innovative method for oil waste elimination. In the present study, fifteen yeast isolates were screened for their lipid content, three of which were chosen for their high lipid content as compared to the standard strain Phaffia rhodozyma NRRL-Y-10921. The three selected isolates were further screened for their fatty acid profile. Yeast isolate (NC-I), identified as Yarrowia lipolytica, was chosen because it exceeded the lipid production of the standard strain by 21%, it also produced the highest C 14:0 (myristic acid), C 18:1 (oleic acid) and C18:2 (linoleic acid), compared to the other two isolates. Growth on different oil wastes resulted in an increase in total lipid content which reached its maximum when oil waste of frying vegetables was added to the media (57.89%). A variation in the fatty acid profile was detected when different types of oil waste were used before and after fermentation. The addition of different glucose concentrations to the vegetable oil waste media resulted in the appearance of C 22:0 (behenic acid) which was not present when the basal medium was used. Scanning Electron Microscopy indicated morphologic changes when the yeast was grown in high glucose concentration as compared to those grown in oil waste media. The activity of malate dehydrogenase (MDH) enzyme exhibited a correlative relationship with the lipid content under various glucose concentrations. The obtained results indicate that vegetable oil waste is suitable for microbial fatty acid production and that the fatty acid profile could be maneuvered through the manipulation of the fermentation media.