Sequential analysis of global gene expression profiles in immature and in vitro matured bovine oocytes: potential molecular markers of oocyte maturation

Background: Without intensive selection, the majority of bovine oocytes submitted to in vitro embryo production (IVP) fail to develop to the blastocyst stage. This is attributed partly to their maturation status and competences. Using the Affymetrix GeneChip Bovine Genome Array, global mRNA expression analysis of immature (GV) and in vitro matured (IVM) bovine oocytes was carried out to characterize the transcriptome of bovine oocytes and then use a variety of approaches to determine whether the observed transcriptional changes during IVM was real or an artifact of the techniques used during analysis. Results: 8489 transcripts were detected across the two oocyte groups, of which similar to 25.0% (2117 transcripts) were differentially expressed (p < 0.001); corresponding to 589 over-expressed and 1528 under-expressed transcripts in the IVM oocytes compared to their immature counterparts. Over expression of transcripts by IVM oocytes is particularly interesting, therefore, a variety of approaches were employed to determine whether the observed transcriptional changes during IVM were real or an artifact of the techniques used during analysis, including the analysis of transcript abundance in oocytes in vitro matured in the presence of a-amanitin. Subsets of the differentially expressed genes were also validated by quantitative real-time PCR (qPCR) and the gene expression data was classified according to gene ontology and pathway enrichment. Numerous cell cycle linked (CDC2, CDK5, CDK8, HSPA2, MAPK14, TXNL4B), molecular transport (STX5, STX17, SEC22A, SEC22B), and differentiation (NACA) related genes were found to be among the several over-expressed transcripts in GV oocytes compared to the matured counterparts, while ANXA1, PLAU, STC1and LUM were among the over-expressed genes after oocyte maturation. Conclusion: Using sequential experiments, we have shown and confirmed transcriptional changes during oocyte maturation. This dataset provides a unique reference resource for studies concerned with the molecular mechanisms controlling oocyte meiotic maturation in cattle, addresses the existing conflicting issue of transcription during meiotic maturation and contributes to the global goal of improving assisted reproductive technology.

Massive infestation by Perulernaea gamitanae (Crustacea: Cyclopoida: Lernaidae) in juvenile gamitana, cultured in the Peruvian Amazon

The gamitana is a species of socio-economic importance in the Peruvian Amazon, often intensively produced locally for human consumption. Because of this, more studies concerning parasite populations affecting this species culture are necessary. In this study, a heavy copepod infestation of Perulernaea gamitanae is reported in a managed culture of gamitana. The prevalence of infection was 100% and mortality of the fish population was complete. The average intensity and abundance of the parasite was 268.8 parasites per individual.

On molecular chirality within naturally occurring secondary organic aerosol particles from the central Amazon Basin

In this perspectives article, we reflect upon the existence of chirality in atmospheric aerosol particles. We then show that organic particles collected at a field site in the central Amazon Basin under pristine background conditions during the wet and dry seasons consist of chiral secondary organic material. We show how the chiral response from the aerosol particles can be imaged directly without the need for sample dissolution, solvent extraction, or sample preconcentration. By comparing the chiral-response images with optical images, we show that chiral responses always originate from particles on the filter, but not all aerosol particles produce chiral signals. The intensity of the chiral signal produced by the size resolved particles strongly indicates the presence of chiral secondary organic material in the particle. Finally, we discuss the implications of our findings on chiral atmospheric aerosol particles in terms of climate-related properties and source apportionment.

Nonsingular bounce in modified gravity

We investigate bouncing solutions in the framework of the nonsingular gravity model of Brandenberger, Mukhanov and Sornborger. We show that a spatially flat universe filled with ordinary matter undergoing a phase of contraction reaches a stage of minimal expansion factor before bouncing in a regular way to reach the expanding phase. The expansion can be connected to the usual radiation-and matter-dominated epochs before reaching a final expanding de Sitter phase. In general relativity (GR), a bounce can only take place provided that the spatial sections are positively curved, a fact that has been shown to translate into a constraint on the characteristic duration of the bounce. In our model, on the other hand, a bounce can occur also in the absence of spatial curvature, which means that the time scale for the bounce can be made arbitrarily short or long. The implication is that constraints on the bounce characteristic time obtained in GR rely heavily on the assumed theory of gravity. Although the model we investigate is fourth order in the derivatives of the metric (and therefore unstable vis-a-vis the perturbations), this generic bounce dynamics should extend to string-motivated nonsingular models which can accommodate a spatially flat bounce.

Structural, dielectric, and optical properties of yttrium calcium borate glasses

Structural and optical properties of stable glasses in the Y(2)O(3)-CaO-B(2)O(3) system, containing the same Y/Ca ratio as the YCa(4)O(BO(3))(3) (YCOB) crystal, were determined from Raman and reflectance infrared spectroscopy. Changes in optical functions with composition are associated with an increase in the number of non-bridging oxygen and to calcium/yttrium oxides content. Refractive indexes values (from 1.597 to 1.627 at lambda=2 mu m) are in good agreement with those of the YCOB crystal, an indication that these glasses are potential candidates for optical applications due to their ease of shaping as large bulk samples or fibers.

Changes in wood-water relationship due to heat treatment assessed on micro-samples of three Eucalyptus species

Wood-water relationship of untreated and heat-treated wood was studied. Specimens of Eucalyptus grandis, E saligna, and E citriodora were submitted to five conditions of heat treatment: 180 degrees C and 220 degrees C with air; 220 degrees C, 250 degrees C, and 280 degrees C with N(2). The wood-water relationships were accurately studied in a special device, in which the moisture content (MC) of the sample was measured with a highly sensitive electronic microbalance placed in a climatic chamber. The dimensions of the sample were collected continuously without contact by means of two high-speed laser scan micrometers. Sorption curves and shrinkage-MC relationships were observed. To study the effects of heat treatment, the following parameters were also determined: fiber saturation point (FPS), wood anisotropy (T/R ratio), shrinkage slope, reduction in hygroscopicity, and anti-shrink efficiency (ASE). The physical properties were significantly affected only at 220 degrees C and above. At heat temperature levels higher than 220 degrees C, the reduction in hygroscopicity and ASE are higher than 40% and continue to be reduced with increasing temperature level. This work also demonstrates that heat treatment does not change the slope of the curves shrinkage vs. MC, proving that heat treatment affects the domain of alterations in wood properties, but not the behavior within this domain.

Description of a new Tetranychus (Acarina, Prostigmata : Tetranychidae) pest of Musa from French Guiana

A new species of tetranychid mite, genus Tetranychus Dufour, 1832, is described and illustrated herein from neotropical area. Tetranychus musae sp. nov. (Acari, Prostigmata: Tetranychidae) differs from other species in the genus by the combination between the arrangement of leg setae on females tarsus I and the shape of the aedeagus. Tarsus I bears one tactile setae proximal to proximal duplex setae and three tactiles in line or almost in line with proximal duplex setae. The aedeagus knob consists of an acute posterior projection bent ventrally and a larger anterior rounded projection directed anterodorsally. T. musae specimens were collected in French Guiana where they appeared to be a pest of Musa sp. A key to adults of neotropical species of the genus Tetranychus feeding on banana is provided.

Biochemical and biopharmaceutical properties of PEGylated uricase

PEGylation is a successful strategy for improving the biochemical and biopharmaceutical properties of proteins and peptides through the covalent attachment of polyethylene glycol chains. In this work, purified recombinant uricase from Candida sp. (UC-r) was modified by PEGylation with metoxypolyethilenoglycol-p-nitrophenyl-carbonate (mPEG-pNP) and metoxypolyethyleneglycol-4,6-dichloro-s-triazine (mPEG-CN). The UC-r-mPEG-pNP and UC-r-mPEG-CN conjugates retained 87% and 75% enzyme activity respectively. The K(M) values obtained 2.7 x 10(-5) M (mPEG-pNP) or 3.0 x 10(-5) M (mPEG-CN) lot the conjugates as compared to 5.4 x 10(-5) M for the native UC-r, suggesting enhancement in the substrate affinity of the enzyme attached. The effects of pH and temperature on PEGylated UC-r indicated that the conjugates were more active at close physiological pH and were stable up to 70 degrees C. Spectroscopic study performed by circular dichroism at 20 degrees C and 50 degrees C did not show any relevant difference in protein structure between native and PEGylated UC-r. In rabbit and Balb/c mice, the native UC-r elicited an intense immune response being highly immunogenic. On the other hand, the PEGylated UC-r when injected chronically in mice did not induce any detectable antibody response. This indicates sufficient reduction of the immunogenicity this enzyme by mPEG-pNP or mPEG-CN conjugation, making it suitable for a possible therapeutical use. (C) 2009 Elsevier B.V. All rights reserved.

Capillary electrophoresis method for plasmatic determination of imatinib mesylate in chronic myeloid leukemia patients

Imatinib (IMAT) is a tyrosine kinase inhibitor that has been used for the treatment of chronic myeloid leukemia (CML). Despite the efficacy of IMAT therapy, some cases of treatment resistance have been described in CML. Developing a plasma method is important since there are several studies that provided a higher correlation between IMAT plasma concentration and response to treatment. Therefore, in this investigation we validated a method by CE as an alternative, new, simple and fast electrophoretic method for IMAT determination in human plasma. The analysis was performed using a fused silica capillary (50 mm id x 46.5 cm total length, 38.0 cm effective length); 50 mmol/L sodium phosphate buffer, pH 2.5, as BGE; hydrodynamic injection time of 20 s (50 mbar); voltage of 30 kV; capillary temperature of 35 degrees C and detection at 200 nm. Plasma samples pre-treatment involved liquid-liquid extraction with methyl-tert-butyl ether as the extracting solvent. The method was linear from 0.125 to 5.00 mg/mL. The LOQ was 0.125 mg/mL. Mean absolute recovery of IMAT was 67%. The method showed to be precise and accurate with RSD and relative error values lower than 15%. Furthermore, the application of the method was performed in the analysis of plasma samples from CML patients undergoing treatment with IMAT.

Glazing to entry, Mt. Nebo House

Leadlight windows with stained glass feature panes and timber muntins over french doors with matching leadlight sidelights.

Dining room, Mt. Nebo House

Interior view showing fireplace and mezzanine.

Hallway, Mt. Nebo House

Hallway off main entry to house.

Dormer window, Mt. Nebo House

Arched head leadlight and casement windows with stained glass feature panes and timber muntins. View through window from interior.

Dormer window, Mt. Nebo House

Arched head leadlight window with stained glass feature panes and timber muntins.

Garage-studio interior, Mt. Nebo House

Timber framing and roof skylight detail inside garage-studio.