Increased frequency of HLA-DRwl4b(w6)-associated RFLP in Greenlanders of Eskimo origin

HLA-DR typing by the restriction fragment length polymorphism (RFLP) technique of 42 Greenlanders living in northern Jutland, Denmark, revealed a phenotype frequency of 33.3% for HLA-DRw14b(w6), which is significantly different from the frequency of 0.0% observed among 98 Danes of Caucasian origin. When comparing the two populations, the frequency of other HLA-DRB allogenotypes show insignificant variations. Since HLA-DRw14b(w6) is carried by approximately one-third of the Greenlanders tested, this allogenotype may serve as a useful marker in further anthropological and immunogenetic studies.     

Toxicity of the blue-green alga (cyanobacterium) Microcystis aeruginosa in drinking water to growing pigs,as an animal model for human injury and risk assessment

Hepatotoxins from blue-green algae are increasingly recognized as a potential hazard in drinking water supplies. The clinical consequences of ingestion include acute or chronic liver injury, with the possibility of enhanced susceptibility to, and growth of, liver tumors. To establish guidelines for water safety requires the demonstration of dose-dependent effects of toxicity and experimental determination of maximum “no-adverse-effect levels.” This paper describes the use of growing pigs as a model for human injury resulting from Microcystis toxins in drinking water. Risk assessment calculations using a series of safety factors are carried out, resulting in a guideline level after incorporating an additional safety factor for tumor promotion of approximately 1.0 μg toxins/L. With the Microcystis used for this trial, that concentration corresponds to 5000 cells/mL. © 1994 by John Wiley & Sons, Inc..     

Fibrinolytic capacity in hemodialysis patients treated with recombinant human erythropoietin.

A major adverse effect of recombinant human erythropoietin (r-HuEPO) in hemodialyzed patients are thrombotic events. Several reports on platelet function during r-HuEPO treatment have been published but less is known about fibrinolysis. In the present study, the fibrinolytic capacity was studied in 20 patients on maintenance hemodialysis and treated with r-HuEPO. The patients were randomized into two groups and investigated in a crossover design. r-HuEPO was administered intravenously and subcutaneously in each group and was given for 3 months, respectively. Plasma tissue plasminogen activator (t-PA) and released t-PA remained unaffected by r-HuEPO in both groups throughout the study. Tissue plasminogen activator inhibitor (PAI) increased in a cyclic way reaching peak values 4-6 weeks after the start of investigation and again 4-6 weeks after changing therapy. The increase in PAI was significant in the two groups (0.025 > p > 0.01). Tissue plasminogen antigen was low in the uremic patients. The influence of r-HuEPO on this parameter was not investigated. Compensatory changes in plasma levels of factor XII procoagulant activity, activated protein C and of alpha 2-antiplasmin were not observed. Thrombotic events occurred in 4 patients at peak values of PAI. Six patients required an increase in heparin dose simultaneously with the increase in PAI. Thus, r-HuEPO seemed to affect the fibrinolytic capacity of uremic patients.     

Rapid genotyping of MBL2 gene mutations using real-time PCR with fluorescent hybridisation probes

In this study, we describe a real-time polymerase chain reaction (PCR) for genotyping all known polymorphisms of the human mannose-binding lectin 2 (MBL2) gene. These comprised two variations in the 5' regulatory region at positions -550 (H/L) and -221 (X/Y), one in the 5' untranslated sequence at position +4 (P/Q) and three structural mutations within exon 1 at codons 52, 54, and 57, also known as the D, B and C variants, respectively. Three reactions with two different conditions were sufficient to genotype one individual unambiguously. The three mutations in exon 1 were detected in one capillary using a sensor probe covering the three mutations, whereas amplification of the variants located upstream of the coding sequence was performed in only two reactions. Single colour detection was used for detection of the (H/L) polymorphism and multiplexing by dual colour probes was used for simultaneous genotyping of (X/Y) and (P/Q). The reliability of the system was evaluated by comparison with a conventional PCR method with sequence-specific primers (PCR-SSP). For this study, 100 individuals of Danish and 30 of African descent were analysed, and the genotypes obtained were concordant in all cases. This new method is rapid and provides reliable results without ambiguities.     

Optimisation of radiographic acquisition parameters for direct digital radiography: A systematic review

IntroductionThe objective of this systematic review was to uncover and synthesise all available literature regarding appropriate acquisition parameters for direct digital radiography. It sought to either confirm current practices as optimal, or to uncover practices that may produce more optimised results.MethodsA comprehensive search of published and unpublished literature was undertaken to find studies that evaluated how adjustment of different acquisition parameters affected subjective image quality and patient radiation dose. Eight hundred and fifty-eight studies were retrieved for title and abstract screening. Eighty-nine studies were retrieved for full-text screening, and 23 were included for review and methodological quality screening.ResultsNarrative synthesis of the 23 included studies revealed limited evidence to guide any potential change or acceptance of currently accepted best practice. Meta-analysis was unable to be performed for any of the included studies due to high levels of methodological heterogeneity. A key finding of this review was that the goals of optimisation research varied greatly across the included studies.ConclusionSignificant methodological heterogeneity in the included studies limited the number of clinically relevant findings that would give evidence to an acceptance of, or suggest changes to, currently accepted best practice. Improving consistency in approach across future works of technique optimisation will ensure future systematic reviews will be able to provide strong evidence and meta-analysis will be able to be performed.Implications for clinical practiceThis review highlights that in the literature, studies of optimisation of radiographic acquisition parameters have varying goals. This methodological heterogeneity limits the applicability of systematic reviews and precludes the use of meta-analysis. The authors recommend that a framework for optimisation research be produced as a priority to help improve homogeneity in future research.     

Functional characterization of BRCA1 gene variants by mini-gene splicing assay

Mutational screening of the breast cancer susceptibility gene BRCA1 leads to the identification of numerous pathogenic variants such as frameshift and nonsense variants, as well as large genomic rearrangements. The screening moreover identifies a large number of variants, for example, missense, silent, and intron variants, which are classified as variants of unknown clinical significance owing to the lack of causal evidence. Variants of unknown clinical significance can potentially have an impact on splicing and therefore functional examinations are warranted to classify whether these variants are pathogenic or benign. Here we validate a mini-gene splicing assay by comparing the results of 24 variants with previously published data from RT-PCR analysis on RNA from blood samples/lymphoblastoid cell lines. The analysis showed an overall concordance of 100%. In addition, we investigated 13 BRCA1 variants of unknown clinical significance or putative variants affecting splicing by in silico analysis and mini-gene splicing assay. Both the in silico analysis and mini-gene splicing assay classified six BRCA1 variants as pathogenic (c.80+1G>A, c.132C>T (p.=), c.213−1G>A, c.670+1delG, c.4185+1G>A, and c.5075−1G>C), whereas six BRCA1 variants were classified as neutral (c.-19-22_-19-21dupAT, c.302−15C>G, c.547+14delG, c.4676−20A>G, c.4987−21G>T, and c.5278−14C>G) and one BRCA1 variant remained unclassified (c.670+16G>A). In conclusion, our study emphasizes that in silico analysis and mini-gene splicing assays are important for the classification of variants, especially if no RNA is available from the patient. This knowledge is crucial for proper genetic counseling of patients and their family members.     

Emerging role of thyroid hormone metabolites

Thyroid hormones (THs) are essential for the regulation of development and metabolism in key organs. THs produce biological effects both by directly affecting gene expression through the interaction with nuclear receptors (genomic effects) and by activating protein kinases and/or ion channels (short‐term effects). Such activations can be either direct, in the case of ion channels, or mediated by membrane or cytoplasmic receptors. Short‐term‐activated signalling pathways often play a role in the regulation of genomic effects. Several TH intermediate metabolites, which were previously considered without biological activity, have now been associated with a broad range of actions, mostly attributable to short‐term effects. Here, we give an overview of the physiological roles and mechanisms of action of THs, focusing on the emerging position that TH metabolites are acquiring as important regulators of physiology and metabolism.