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.     

CXCL12 and TP53 genetic polymorphisms as markers of susceptibility in a Brazilian children population with acute lymphoblastic leukemia (ALL)

Acute lymphoblastic leukemia (ALL) is the most common pediatric malignancy. Genetic polymorphisms in the 3′UTR region of the CXCL12 (rs1801157) and TP53 codon 72 (rs1042522) genes may contribute to susceptibility to childhood ALL because they affect some important processes, such as metastasis regulation and tumor suppression. Thus the objective of the present study was to detect the frequency of two genetic polymorphisms in ALL patients and controls and to add information their impact on genetic susceptibility and prognosis. The CXCL12 and TP53 polymorphisms were tested in 54 ALL child patients and in 58 controls by restriction fragment length polymerase chain reaction and allelic specific chain reaction techniques, respectively. The frequencies of both allelic variants were higher in ALL patients than in the controls and indicated a positive association: OR = 2.44; 95 % CI 1.05–5.64 for CXCL12 and OR = 2.20; 95 % CI 1.03–4.70 for TP53. Furthermore, when the two genetic variants were analyzed together, they increased significantly more than fivefold the risk of this neoplasia development (OR = 5.24; 95 % CI 1.39–19.75), indicating their potential as susceptibility markers for ALL disease and the relevance of the allelic variant combination to increased risk of developing malignant tumors. Future studies may indicate a larger panel of genes involved in susceptibility of childhood ALL and other hematological neoplasias.     

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.     

Internal efficiency, nutrient uptake, and the relation to field water resources in rainfed lowland rice of northeast Thailand

Rice-based (Oryza sativa L.) rainfed lowlands are the major cropping system in northeast Thailand. Average yields are low, which is generally explained by frequent drought events, low soil fertility, and poor fertilizer response. However, neither the relative importance of these factors nor their interaction is well understood. Therefore, we analyzed an existing database on fertilizer trials conducted between 1995 and 1997 at eight different sites in northeast Thailand with the objective to determine indigenous nutrient supplies, internal efficiencies, and recovery efficiencies of applied nutrients in rainfed lowland rice. Of particular interest was the effect of variety type (traditional) and water supply on these components. Comparison of N, P, and K concentrations in grain and straw (average N–P–K grain concentration of 11.0–2.7–3.4 g kg⁻¹; average N–P–K straw concentration of 5.2–0.9–16.4 g kg⁻¹) in the traditional-type varieties used at all trial sites with literature values showed no differences for these parameters between traditional and modern-type varieties or between irrigated and rainfed environments. In contrast, internal efficiencies of N, P, and K (average IEN: 46 kg grain per kg N uptake; IEP: 218 kg grain per kg P uptake; IEK: 25 kg grain per kg K uptake) were much lower than reported for irrigated systems, and the difference was greatest for K, which is mainly accumulated in the straw. Indigenous nutrient supply (average INS: 38 kg ha⁻¹; IPS: 10 kg ha⁻¹; IKS: 89 kg ha⁻¹) and recovery efficiency (average REN: 0.28 kg kg⁻¹; REP: 0.13 kg kg⁻¹; REK: 0.49 kg kg⁻¹) were low but comparable to the lower values reported from irrigated systems. Average seasonal field water resources seemed to reduce the indigenous nutrient supply but had no or little effect on internal efficiency and recovery efficiency. We concluded that the main reason for the low system productivity without and with fertilizer in northeast Thailand is the dominant use of traditional-type varieties with low harvest indices, which was the dominant cause for the observed low internal nutrient efficiency. Therefore, intensification of rainfed systems through substantially increased nutrient inputs can be recommended only where varieties with an average harvest index of close to 0.4 or higher are available.