Microwave sterilization of plastic tissue culture vessels for reuse

A simple protocol has been developed for recycling plastic tissue culture vessels. The killing properties of microwaves were used to decontaminate plastic tissue culture vessels for reuse. Nine bacterial cultures, four gram-negative and five gram-positive genera, including two Bacillus species, were used to artificially contaminate tissue culture vessels. The microwaves produced by a "home-type" microwave oven (2.45 gHz) were able to decontaminate the vessels with a 3-min exposure. The same exposure time was also used to completely inactivate the following three test viruses: polio type 1, parainfluenza type 1 (Sendai), and bacteriophage T4. The recycling procedure did not reduce the attachment and proliferation of the following cell types: primary chicken and turkey embryo, HEp-2, Vero, BGMK, and MK-2.Peer reviewedMicrobiolog

Increased snow depth affects microbial activity and nitrogen mineralization in two Arctic tundra communities

Microbial activity in Arctic tundra ecosystems continues through the winter and is an important component of the annual C budget. This activity is sensitive to climatic variation, particularly snow depth because that regulates soil temperature. The influence of winter conditions on soil N cycling is poorly understood. In this study, we used intact core incubations sampled periodically through the winter and following growing season to measure net N mineralization and nitrification in dry heath and in moist tussock tundra under ambient and experimentally increased snow depths (by use of a snowfence). In dry heath, we sampled soils under Dryas octopetela or Arctostaphylos alpine, while in tussock tundra, we sampled Eriophorum vaginatum tussocks and Sphagnum dominated areas between tussocks. Our objectives were to: (1) examine how different winter snow regimes influenced year-round N dynamics in the two tundra types, and (2) evaluate how these responses are affected by dominant species present in each system. In tussock tundra, soils with increased winter snow cover had high net N mineralization rates during the fall and winter, followed by immobilization during thaw. In contrast, N mineralization only occurred during the autumn in soils with ambient snow cover. During the growing season when N immobilization dominated in areas with ambient snow cover, soils with increased winter snow cover had positive net mineralization and nitrification rates. In dry heath tundra, soils with increased snow depth had high late winter net N mineralization rates, but these rates were: (a) comparable to early winter rates in soils under Arctostaphylos plants with ambient snow cover; (b) greater in soils under Arctostaphylos plants than in soils under Dryas plants; and (c) less than the rates found in tussock tundra. Our findings suggest under ambient snow conditions, low soil temperatures limit soil N mineralization, but that deeper snow conditions with the associated warmer winter soil temperatures dramatically increase over-winter N mineralization and thereby alter the amount and timing of plant-available N in tundra ecosystems. (C) 2003 Elsevier Ltd. All rights reserved

The chemical synthesis, stability, and activity of MAIT cell prodrug agonists that access MR1 in recycling endosomes

Mucosal-associated invariant T (MAIT) cells are antibacterial effector T cells that react to pyrimidines derived from bacterial riboflavin synthesis presented by the monomorphic molecule MR1. A major challenge in MAIT cell research is that the commonly used MAIT agonist precursor, 5-amino-6-d-ribitylaminouracil (5-A-RU), is labile to autoxidation, resulting in a loss of biological activity. Here, we characterize two independent autoxidation processes by LCMS. To overcome the marked instability, we report the synthesis of a 5-A-RU prodrug generated by modification of the 5-amino group with a cleavable valine-citrulline-p-aminobenzyl carbamate. The compound is stable in prodrug form, with the parent amine (i.e., 5-A-RU) released only after enzymatic cleavage. Analysis of the prodrug in vitro and in vivo showed an enhanced MAIT cell activation profile compared to 5-A-RU, which was associated with preferential loading within recycling endosomes, a route used by some natural agonists. This prodrug design therefore overcomes the difficulties associated with 5-A-RU in biological studies and provides an opportunity to explore different presentation pathways

Comparison of the performance of the IDEXX SediVue Dx® with manual microscopy for the detection of cells and 2 crystal types in canine and feline urine

Background Microscopic evaluation of urine is inconsistently performed in veterinary clinics. The IDEXX SediVue Dx® Urine Sediment Analyzer (SediVue) recently was introduced for automated analysis of canine and feline urine and may facilitate performance of urinalyses in practice. Objective Compare the performance of the SediVue with manual microscopy for detecting clinically relevant numbers of cells and 2 crystal types. Samples Five‐hundred thirty urine samples (82% canine, 18% feline). Methods For SediVue analysis (software versions [SW] 1.0.0.0 and 1.0.1.3), uncentrifuged urine was pipetted into a cartridge. Images were captured and processed using a convolutional neural network algorithm. For manual microscopy, urine was centrifuged to obtain sediment. To determine sensitivity and specificity of the SediVue compared with manual microscopy, thresholds were set at ≥5/high power field (hpf) for red blood cells (RBC) and white blood cells (WBC) and ≥1/hpf for squamous epithelial cells (sqEPI), non‐squamous epithelial cells (nsEPI), struvite crystals (STR), and calcium oxalate dihydrate crystals (CaOx Di). Results The sensitivity of the SediVue (SW1.0.1.3) was 85%‐90% for the detection of RBC, WBC, and STR; 75% for CaOx Di; 71% for nsEPI; and 33% for sqEPI. Specificity was 99% for sqEPI and CaOx Di; 87%‐90% for RBC, WBC, and nsEPI; and 84% for STR. Compared to SW1.0.0.0, SW1.0.1.3 had increased sensitivity but decreased specificity. Performance was similar for canine versus feline and fresh versus stored urine samples. Conclusions and Clinical Importance The SediVue exhibits good agreement with manual microscopy for the detection of most formed elements evaluated, but improvement is needed for epithelial cells