A benign approach to the preparation of freshwater bryozoan statoblasts for scanning electron microscopy (SEM) imaging

Abstract

Several different species of freshwater Bryozoa, belonging to the genera Plumatella, Rumarcanella and Fredericella, were detected within the Northern Mallee Pipeline (NMP) system in Victoria, Australia, that required definitive identification. These organisms produce asexual buds called statoblasts, with valves composed of sclerotised chitin that bear minute micro-ornamentations of considerable taxonomical significance. Imaging and analysis of these distinctive micro-ornamentations using scanning electron microscopy (SEM) is often employed for species identification. Meticulous preparation of statoblast samples is therefore required that necessitates the removal of adhering debris, dehydration and drying—whilst mitigating specimen damage and distortion. This technical note describes an approach whereby each of these three steps have been individually designed to be as benign as possible, using mild detergent/sonication to remove debris, a gradual and gentle dehydration procedure using ethanol, and critical point drying. For the overall process, these methods are chosen to optimise control and to minimise the use of harsh and hazardous chemicals.     

The use of adjunctive GPIIb/IIIa inhibitors in patients with unstable angina/non-Q-wave MI undergoing percutaneous coronary intervention

Glycoprotein IIb/IIIa receptor inhibitors represent a relatively new therapeutic approach in the field of antiplatelet therapy. Following the development of abciximab a number of small molecule GPIIb/IIIa inhibitors have been introduced such as tirofiban and eptifibatide. In this fast-moving field the interventional cardiologist needs a framework to guide decision-making for the individual patient. This review covers the efficacy and safety data from the clinical trials of GPIIb/IIIa inhibitors in the context of patients undergoing percutaneous coronary intervention for unstable angina/non-Q-wave myocardial infarction. There is an increasing body of evidence to support the efficacy of GPIIb/IIIa inhibitors in reducing the risk of adverse ischemic events in high and low risk patients undergoing percutaneous coronary intervention. A number of unresolved efficacy and safety issues remain, including the duration of treatment before and after intervention; whether a reduction in the heparin dose would further decrease the risk of hemorrhage without affecting the periprocedural thrombotic rate in patients undergoing PTCA with adjunctive GPIIb/IIIa inhibitors; and the cost-effectiveness of this therapy. When a thorough analysis of cost-effectiveness has been made, it will be easier to advocate the widespread use of these agents in all patients undergoing coronary intervention.     

A high-throughput cloning system for reverse genetics in <Emphasis Type="Italic">Trypanosoma cruzi</Emphasis>

Background  

The three trypanosomatids pathogenic to men, Trypanosoma cruzi, Trypanosoma brucei and Leishmania major, are etiological agents of Chagas disease, African sleeping sickness and cutaneous leishmaniasis, respectively. The complete sequencing of these trypanosomatid genomes represented a breakthrough in the understanding of these organisms. Genome sequencing is a step towards solving the parasite biology puzzle, as there are a high percentage of genes encoding proteins without functional annotation. Also, technical limitations in protein expression in heterologous systems reinforce the evident need for the development of a high-throughput reverse genetics platform. Ideally, such platform would lead to efficient cloning and compatibility with various approaches. Thus, we aimed to construct a highly efficient cloning platform compatible with plasmid vectors that are suitable for various approaches.     

Long-term population trends of endangered Hawaiian waterbirds

We analyzed long-term winter survey data (1956–2007) for three endangered waterbirds endemic to the Hawaiian Islands, the Hawaiian moorhen (Gallinula chloropus sandvicensis), Hawaiian coot (Fulica alai), and Hawaiian stilt (Himantopus mexicanus knudseni). Time series were analyzed by species–island combinations using generalized additive models, with alternative models compared using Akaike information criterion (AIC). The best model included three smoothers, one for each species. Our analyses show that all three of the endangered Hawaiian waterbirds have increased in population size over the past three decades. The Hawaiian moorhen increase has been slower in more recent years than earlier in the survey period, but Hawaiian coot and stilt numbers still exhibit steep increases. The patterns of population size increase also varied by island, although this effect was less influential than that between species. In contrast to earlier studies, we found no evidence that rainfall affects counts of the target species. Significant population increases were found on islands where most wetland protection has occurred (Oahu, Kauai), while weak or no increases were found on islands with few wetlands or less protection (Hawaii, Maui). Increased protection and management, especially on Maui where potential is greatest, would likely result in continued population gains, increasing the potential for meeting population recovery goals.     

Immunoprophylaxis of respiratory syncytial virus: global experience

In sarcoidosis, host genetic factors are discussed as contributing to disease susceptibility and course. Since tumor necrosis factor (TNF)-α is a central mediator of granuloma formation and since elevated TNF-α levels are found during active phases of sarcoidosis, genetic polymorphisms correlating with influences on TNF-α levels are of special interest. The complete sequencing of the MHC region and the increase in the number of identified gene polymorphisms in this locus associated with TNF-α production offer the opportunity of detecting new genes associated with sarcoidosis and perhaps of defining disease-associated haplotypes that bear the potential of serving as predictive markers for this disease.     

Patterns of investment in reproductive and somatic tissues in the loliginid squid <Emphasis Type="Italic">Loligo forbesii</Emphasis> and <Emphasis Type="Italic">Loligo vulgaris</Emphasis> in Iberian and Azorean waters

The veined squid, Loligo forbesii, is found throughout the northeast Atlantic, including the waters off the Iberian Peninsula, and is a socio-economically important cephalopod species, sustaining several small-scale commercial and local artisanal fisheries. This study uses Iberian and Azorean trawl survey and fishery landings data from 1990 to 1992 and employs additive modelling to examine the relationships amongst somatic growth, season and gonad growth, in an attempt to determine the relative importance of intrinsic (e.g. nutritional state and body size) and extrinsic (temperature and daylight) factors which contribute to maturation in L. forbesii. We compare the results with those from a comparative analysis of contemporaneous data on Loligo vulgaris from the Iberian coast, and with a re-analysis of previously published results for L. forbesii in Scottish waters. Reproductive organ weight in both sexes of L. forbesii from all ports shows seasonal patterns with a fall/winter peak in maturation, as is expected with the animals’ year-long life cycle. Two maturity peaks (spring and winter) were found for both Azores L. forbesii males and females, possibly evidence of two breeding cohorts, with males maturing approximately 1 month earlier than females in both seasons. For L. vulgaris, three annual peaks in reproductive weight were identified. With the exception of L. forbesii males from Scottish waters, in all study animals, the partial effect of mantle length and mantle weight on reproductive weight is positive up to large body sizes. The relationship between nutritional status and reproductive weight is also generally positive. Geographic variation in the relationships, as seen between Scottish males and samples from the other study areas, is hypothesised to be largely attributable to local differences in environmental conditions (productivity, food availability, temperature, upwelling), although differences between L. forbesii from the Azores, Scotland and mainland Europe may also relate to previously documented genetic differences.     

Flexibility in crosstalk between H2B ubiquitination and H3 methylation in vivo

Histone H2B ubiquitination is a dynamic modification that promotes methylation of histone H3K79 and H3K4. This crosstalk is important for the DNA damage response and has been implicated in cancer. Here, we show that in engineered yeast strains, ubiquitins tethered to every nucleosome promote H3K79 and H3K4 methylation from a proximal as well as a more distal site, but only if in a correct orientation. This plasticity indicates that the exact location of the attachment site, the native ubiquitin-lysine linkage and ubiquitination cycles are not critical for trans-histone crosstalk in vivo. The flexibility in crosstalk also indicates that other ubiquitination events may promote H3 methylation.