Pre-ovulatory follicular characteristics and ovulation rates in different breed crosses, carriers or non-carriers of the Booroola or Cambridge fecundity gene

Terminal follicular dynamics and ovulation rates (OR) were compared in different local breeds after introducing fecundity genes of different origin. Crossbred ewes which were carriers (F+) or non-carriers (++) of Booroola (BFec) or Cambridge genes (CFec) were included: CambridgexCambridge (CC), CambridgexSuffolk (CS), CambridgexTexel (CT), BooroolaxTexel (BT) and BooroolaxGerman Mutton Merino (BGM). The numbers of small (diameter 2-3.5 mm), medium (diameter >3.5-5.0 mm) and large (diameter >5.0 mm) growing follicles, the maximum diameter before ovulation and the regression and artesia rates of ovarian follicles >/=2 mm in diameter were studied laparoscopically and repeatedly during the last 5 days of an induced oestrous cycle. The ORs were determined one cycle before and two cycles after the repeated laparoscopy. BFec and CFec significantly enhanced the OR of all crossbreeds. Carriers of BFec or CFec did not have significantly different ORs due to any crossbreeding effect. The same observation was made for non-carriers of both Fec gene types. Whatever the crossbreed, the number of small, medium and large growing follicles were similar between carriers and non-carriers in spite of a higher number of ovulating follicles in carriers of both Fec gene types. The diameter of ovulatory follicles did not differ among crossbreds, or between carriers and non-carriers except in the BT (5.2+/-0.2 vs. 6.5+/-0.8 mm, respectively) and CC (6.6+/-0.2 vs. 5.6+/-0.3 mm) ewes.The higher OR in the presence of the Booroola gene was associated with a low atresia rate of large follicles in all crossbreeds (BT: 52+/-8% (F+) vs. 61+/-7% (++); BGM: 51+/-6% vs. 75+/-5%). The high OR of the carriers of the CFec gene seemed to be associated with a lower number of large growing follicles with a lower (P<0.05) atresia rate as compared with Booroola crossbreeds.In conclusion, follicular features were similar between purebred Cambridge and its crossbred CS and CT. In ewes carrying the BFec or CFec gene, the reduction in follicular atresia seemed to be one of the main follicular features implicated in the higher OR.     

Validation and implementation of an internal standard in comet assay analysis

The comet assay is widely used to detect DNA damage in single cells. However, only moderate attention has been paid to the experimental variability of this assay, especially during electrophoresis. To take into account this variation and to be able to compare measurements from different electrophoretic runs, as would be necessary when large numbers of samples need to be analysed, it is important to integrate an internal standard into the assay. This study presents a first step in the validation and implementation of an internal standard in the alkaline comet assay. Untreated and ethyl methanesulfonate treated cells (K562 human erythroleukemia cell line) were used as negative and positive internal standards, respectively, in each electrophoresis run. Three steps were followed: (1) assessment of the different levels of variability which may influence the damage levels of the internal standards, (2) evaluation of the variability across separate electrophoresis runs on the quantification of DNA damage in the internal standards by three experimenters involved in different studies and (3) proposal of an adequate calculation system to integrate the internal standards into test sample data. The application of the two proposed models to samples from a human biomonitoring study is presented. The model which calibrates the measurements against the negative internal standard is the most useful since this negative standard was the most stable across experiments and among the three experimenters. The percentage of DNA in the tail is the most appropriate parameter to analyse induced DNA damage, because its interelectrophoresis and interexperimenter variation is less pronounced than that of tail length.     

Mathematical simulation of the diel O, S, and C biogeochemistry of a hypersaline microbial mat

The creation of a mathematical simulation model of photosynthetic microbial mats is important to our understanding of key biogeochemical cycles that may have altered the atmospheres and lithospheres of early Earth. A model is presented here as a tool to integrate empirical results from research on hypersaline mats from Baja California Sur (BCS), Mexico into a computational system that can be used to simulate biospheric inputs of trace gases to the atmosphere. The first version of our model, presented here, calculates fluxes and cycling of O(2), sulfide, and dissolved inorganic carbon (DIC) via abiotic components and via four major microbial guilds: cyanobacteria (CYA), sulfate reducing bacteria (SRB), purple sulfur bacteria (PSB) and colorless sulfur bacteria (CSB). We used generalized Monod-type equations that incorporate substrate and energy limits upon maximum rates of metabolic processes such as photosynthesis and sulfate reduction. We ran a simulation using temperature and irradiance inputs from data collected from a microbial mat in Guerrero Negro in BCS (Mexico). Model O(2), sulfide, and DIC concentration profiles and fluxes compared well with data collected in the field mats. There were some model-predicted features of biogeochemical cycling not observed in our actual measurements. For instance, large influxes and effluxes of DIC across the MBGC mat boundary may reveal previously unrecognized, but real, in situ limits on rates of biogeochemical processes. Some of the short-term variation in field-collected mat O(2) was not predicted by MBGC. This suggests a need both for more model sensitivity to small environmental fluctuations for the incorporation of a photorespiration function into the model.     

Microbial lithification in marine stromatolites and hypersaline mats

Lithification in microbial ecosystems occurs when precipitation of minerals outweighs dissolution. Although the formation of various minerals can result from microbial metabolism, carbonate precipitation is possibly the most important process that impacts global carbon cycling. Recent investigations have produced models for stromatolite formation in open marine environments and lithification in shallow hypersaline lakes, which could be highly relevant for interpreting the rock record and searching for extraterrestrial life. Two factors that are controlled by microbial processes and physicochemical characteristics determine precipitation: exopolymeric substances and the saturation index, the latter being determined by the pH, {Ca(2+)} and {CO(3)(2-)}. Here, we evaluate community metabolism in microbial mats and hypothesize why these organosedimentary biofilms sometimes lithify and sometimes do not.     

Template-directed synthesis of acyclic oligonucleotide analogues

Bis-phosphoimidazolides of an analogue of adenosine (in which ribose is replaced by an acyclic chain) and of two related analogues of guanosine undergo oligomerization in the presence of complementary polynucleotide templates. Data on the template- and nontemplate-directed reactions are presented, and the possible relevance to origins of life is discussed.     

Development and Validation of the Scan of Postgraduate Educational Environment Domains (SPEED): A Brief Instrument to Assess the Educational Environment in Postgraduate Medical Education

Introduction

Current instruments to evaluate the postgraduate medical educational environment lack theoretical frameworks and are relatively long, which may reduce response rates. We aimed to develop and validate a brief instrument that, based on a solid theoretical framework for educational environments, solicits resident feedback to screen the postgraduate medical educational environment quality.

Methods

Stepwise, we developed a screening instrument, using existing instruments to assess educational environment quality and adopting a theoretical framework that defines three educational environment domains: content, atmosphere and organization. First, items from relevant existing instruments were collected and, after deleting duplicates and items not specifically addressing educational environment, grouped into the three domains. In a Delphi procedure, the item list was reduced to a set of items considered most important and comprehensively covering the three domains. These items were triangulated against the results of semi-structured interviews with 26 residents from three teaching hospitals to achieve face validity. This draft version of the Scan of Postgraduate Educational Environment Domains (SPEED) was administered to residents in a general and university hospital and further reduced and validated based on the data collected.

Results

Two hundred twenty-three residents completed the 43-item draft SPEED. We used half of the dataset for item reduction, and the other half for validating the resulting SPEED (15 items, 5 per domain). Internal consistencies were high. Correlations between domain scores in the draft and brief versions of SPEED were high (>0.85) and highly significant (p<0.001). Domain score variance of the draft instrument was explained for ≥80% by the items representing the domains in the final SPEED.

Conclusions

The SPEED comprehensively covers the three educational environment domains defined in the theoretical framework. Because of its validity and brevity, the SPEED is promising as useful and easily applicable tool to regularly screen educational environment quality in postgraduate medical education.