Effect of Oil Overlay on Inhibition Potential of Roscovitine in Sheep Cumulus‐Oocyte Complexes

Inhibitors of cyclin‐dependent kinases, as roscovitine, have been used to prevent the spontaneous resumption of meiosis in vitro and to improve the oocyte developmental competence. In this study, the interference of oil overlay on the reversible arrest capacity of roscovitine in sheep oocytes as well as its effects on cumulus expansion was evaluated. For this, cumulus‐oocyte complexes (COCs) were cultured for 20 h in TCM 199 with 10% foetal bovine serum (Control) containing 75 μm roscovitine (Rosco). Subsequently, they were in vitro matured (IVM) for further 18 h in inhibitor‐free medium with LH and FSH. The culture was performed in Petri dishes under mineral oil (+) or in 96 well plates without oil overlay (?) at 38.5°C and 5% CO₂. At 20 and 38 h, the cumulus expansion and nuclear maturation were evaluated under stereomicroscope and by Hoechst 33342 staining, respectively. No group presented cumulus expansion at 20 h. After additional culture with gonadotrophins, a significant rate of COCs from both Control groups (+/?) exhibited total expansion while in both Rosco groups (+/?) the partial expansion prevailed. Among the oocytes treated with roscovitine, 65.2% were kept at GV in the absence of oil overlay while 40.6% of them reached MII under oil cover (p < 0.05). This meiotic arrest was reversible, and proper meiosis progression also occurred in the Control groups (+/?). So, the culture system without oil overlay improved the meiotic inhibition promoted by roscovitine without affecting the cumulus expansion rate or the subsequent meiosis progression.     

Follow-up of mercury levels in fish,human hair and urine in the Madeira and Tapajós basins,Amazon, Brazil

Informal economy of gold mining has contaminated some important river basins in Amazon. Follow-up studies on critical compartments showed some areas with high Hg levels in fish as well as in human hair samples. Average Hg in piscivorous fish in the Madeira river itself was 846 ppb (N=284) with a maximum of 3921 ppb. Mercury in fish from non polluted areas in this basin shows high variability, even for single species. A seasonal variation in Hg content was observed, with higher values at the end of the dry season. In the upper Tapajós basin comparable values were found for fish but with a definite decreasing trend downstream. Average value for piscivorous fish in the whole Tapajós basin is 482 ppb (N=122) with a maximum value of 3770 ppb. Hair Hg was higher in fishing villages in the Tapajós (average: 17 ppm; with N=432 and maximum value of 176 ppm) than in the Madeira (average: 9 ppm; N=169; maximum 71 ppm), and data from some areas of the Tapajós suggest a decrease with time. Mercury was much higher in urine of goldshop workers in Santarém (low Tapajós) than in Alta Floresta (high Tapajbs) and show a decreasing trend in both cases, probably related to the significant decline in gold mining activities during the study period (1986–1994).     

Major compositional units of the moon: lunar prospector thermal and fast neutrons

Global maps of thermal and fast neutron fluxes from the moon suggest three end-member compositional units. A high thermal and low fast neutron flux unit correlates with the lunar highlands and is consistent with feldspathic rocks. The South Pole-Aitken basin and a strip that surrounds the nearside maria have intermediate thermal and fast neutron flux levels, consistent with more mafic rocks. There appears to be a smooth transition between the most mafic and feldspathic compositions, which correspond to low and high surface altitudes, respectively. The maria show low thermal and high fast neutron fluxes, consistent with basaltic rocks.     

Lunar Fe and Ti abundances: comparison of lunar prospector and clementine data

The Lunar Prospector neutron spectrometer data correlate well with iron and titanium abundances obtained through analysis of Clementine spectral reflectance data. With the iron and titanium dependence removed, the neutron spectrometer data also reveal regions with enhanced amounts of gadolinium and samarium, incompatible rare earth elements that are enriched in the final phases of magma crystallization. These regions are found mainly around the ramparts of the Imbrium impact basin but not around the other basins, including the much larger and deeper South Pole-Aitken basin. This result confirms the compositional uniqueness of the surface and interior of the Imbrium region.     

Organogenesis and foetal haemodynamics during the normal gestation of healthy black‐rumped agoutis (Dasyprocta prymnolopha,Wagler, 1831) bred in captivity

The objective of this study was to define the patterns of organogenesis and foetal haemodynamics during the normal gestation of healthy agoutis (Dasyprocta prymnolopha) kept in captivity. Thirty pregnant agoutis that ranged in size from small to medium and weighed between 2.5 and 3 kg underwent B‐mode and Doppler ultrasonography for the biometric evaluation of the foetal organs. The foetal aortic blood flow proved to be predominantly systolic, and the measured flow velocity was 78.89 ± 2.95 cm/s, with a maximum pressure gradient of 2.12 ± 0.27 mmHg. The liver was characterized by its large volume, occupying the entire cranial aspect of the abdominal cavity, and it was associated cranially with the diaphragm and caudally with the stomach. The flow velocity in the portal vein was estimated to equal 12.17 ± 2.37 cm/s, with a resistivity index of 0.82 ± 0.05. The gallbladder was centrally located and protruded cranially towards the diaphragm. The spleen was visualized as an elongated structure with tapered cranial and caudal extremities, and the foetal kidneys were visualized bilaterally in the retroperitoneal region, with the right kidney positioned slightly more cranially than the left. The morphological characterization and hemodynamic analysis of the foetal organs of black‐rumped agoutis via B‐mode and Doppler ultrasonography allow determination of the vascular network and of reference values for the blood flow required for perfusing the anatomical elements essential for maintaining the viability of foetuses at different gestational ages.     

Focused evolution of HIV-1 neutralizing antibodies revealed by structures and deep sequencing

Antibody VRC01 is a human immunoglobulin that neutralizes about 90% of HIV-1 isolates. To understand how such broadly neutralizing antibodies develop, we used x-ray crystallography and 454 pyrosequencing to characterize additional VRC01-like antibodies from HIV-1-infected individuals. Crystal structures revealed a convergent mode of binding for diverse antibodies to the same CD4-binding-site epitope. A functional genomics analysis of expressed heavy and light chains revealed common pathways of antibody-heavy chain maturation, confined to the IGHV1-2*02 lineage, involving dozens of somatic changes, and capable of pairing with different light chains. Broadly neutralizing HIV-1 immunity associated with VRC01-like antibodies thus involves the evolution of antibodies to a highly affinity-matured state required to recognize an invariant viral structure, with lineages defined from thousands of sequences providing a genetic roadmap of their development.     

Seasonal Fluxes of Particulate Matter in a Small Closed Lake in Northern Estonia

The seasonal fluxes of dry matter, spheroidal fly-ash particles and pollen were studied in a small mesotrophic lake in order to reveal the patterns of their spatial-temporal distribution in regard to seasonal variations. Sediment traps deployed in the lake at different depths and locationswere used to collect samples for calculating sediment fluxes in two years. The results show that resuspension of settled particles in the lake with a small dynamic ratio is moderate and depends on seasonal changes in thermal stratification. The rate of particle deposition during the summer stratification period was low. The weather conditions during early spring, such as air temperature and the thickness of snow-cover, influence the intensity of resuspension. Air temperature and cloudiness during the summer stratification period determine the rate of bioproduction and thermocline parameters, which also have a major effect on the seston composition and fluxes.     

Protection of mice against fatal herpes simplex type 2 infection by liposomes containing muramyl tripeptide

Intravenous administration of liposomes containing muramyl tripeptide phosphatidylethanolamine, a lipophilic derivative of muramyl dipeptide that activates macrophages to a cytolytic state in situ, significantly protected mice against lethal challenge with herpes simplex virus type 2. These findings suggest that the systemic activation of macrophages by liposomes containing an immunomodulator can lead to prophylaxis of severe infections caused by herpesviruses.     

Transition-State Spectroscopy of Cyclooctatetraene

The 351-nanometer photoelectron spectrum of the planar cyclooctatetraene radical anion (COT·-) shows transitions to two electronic states of cyclooctatetraene (COT). These states correspond to the D4h 1A1g state, which is the transition state for COT ring inversion, and the D8h 3A2u state. The electron binding energy of the 1A1g transition state is 1.099 ± 0.010 electron volts, which is lower by 12.1 ± 0.3 kilocalories per mole than that of the 3A2u state. The photoelectron spectrum shows that the singlet lies well below the triplet in D8h COT and confirms ab initio predictions that the molecule violates Hund's rule. Vibrational structure is observed for both features and is readily assigned by use of a simple potential energy surface.