Bar-Code System for a Microbiological Laboratory

A bar-code system has been assembled for a microbiological laboratory that must examine a large number of samples. The system includes a commercial bar-code reader, computer hardware and software components, plus custom-designed database software. The software generates a user-friendly, menu-driven interface

Bioremediation Potential of the Microflora in a Chlorinated Alkene Contaminated Industrial Leachate

Three major microbial subpopulations from an industrial leachate system were characterized with respect to their bioremediation potential, and particular aspects of a cometabolically active subpopulation were determined

Thermal Spore Exposure Vessels

Thermal spore exposure vessels (TSEVs) are laboratory containers designed for use in measuring rates of death or survival of microbial spores at elevated temperatures. A major consideration in the design of a TSEV is minimizing thermal mass in order to minimize heating and cooling times. This is necessary in order to minimize the number of microbes killed before and after exposure at the test temperature, so that the results of the test accurately reflect the effect of the test temperature. A typical prototype TSEV (see figure) includes a flat-bottomed stainless-steel cylinder 4 in. (10.16 cm) long, 0.5 in. (1.27 cm) in diameter, having a wall thickness of 0.010 plus or minus 0.002 in. (0.254 plus or minus 0.051 mm). Microbial spores are deposited in the bottom of the cylinder, then the top of the cylinder is closed with a sterile rubber stopper. Hypodermic needles are used to puncture the rubber stopper to evacuate the inside of the cylinder or to purge the inside of the cylinder with a gas. In a typical application, the inside of the cylinder is purged with dry nitrogen prior to a test. During a test, the lower portion of the cylinder is immersed in a silicone-oil bath that has been preheated to and maintained at the test temperature. Test temperatures up to 220 C have been used. Because the spores are in direct contact with the thin cylinder wall, they quickly become heated to the test temperature

Size uniformity of animal cells is actively maintained by a p38 MAPK-dependent regulation of G1-length

Animal cells within a tissue typically display a striking regularity in their size. To date, the molecular mechanisms that control this uniformity are still unknown. We have previously shown that size uniformity in animal cells is promoted, in part, by size-dependent regulation of G1 length. To identify the molecular mechanisms underlying this process, we performed a large-scale small molecule screen and found that the p38 MAPK pathway is involved in coordinating cell size and cell cycle progression. Small cells display higher p38 activity and spend more time in G1 than larger cells. Inhibition of p38 MAPK leads to loss of the compensatory G1 length extension in small cells, resulting in faster proliferation, smaller cell size and increased size heterogeneity. We propose a model wherein the p38 pathway responds to changes in cell size and regulates G1 exit accordingly, to increase cell size uniformity

Root angle is controlled by EGT1in cereal crops employing anantigravitropic mechanism

Root angle in crops represents a key trait for efficient capture of soil resources. Root angle is determined by competing gravitropic versus anti-gravitropic offset (AGO) mechanisms. Here we report a new root angle regulatory gene termed ENHANCED GRAVITROPISM1 (EGT1) that encodes a putative AGO component, whose loss of function enhances root gravitropism. Mutations in barley and wheat EGT1 genes confer a striking root phenotype, where every root class adopts a steeper growth angle. EGT1 encodes a F-box and Tubby domain containing protein which is highly conserved across plant species. Haplotype analysis found that natural allelic variation at the barley EGT1 locus impacts root angle. Gravitropic assays indicated that Hvegt1 roots bend more rapidly than wildtype. Transcript profiling revealed Hvegt1 roots deregulate ROS homeostasis and cell wall-loosening enzymes and cofactors. ROS imaging shown that Hvegt1 root basal meristem and elongation zone tissues have reduced levels. Atomic Force Microscopy measurements detected elongating Hvegt1 root cortical cell walls are significantly less stiff than wildtype. In situ analysis identified HvEGT1 is expressed in elongating cortical and stele tissues, which are distinct from known root gravitropic perception and response tissues in the columella and epidermis, respectively. We propose that EGT1 controls root angle by regulating cell wall stiffness in elongating root cortical tissue, counteracting the gravitropic machinery’s known ability to bend the root via its outermost tissues. We conclude that root angle is controlled by EGT1 in cereal crops employing a novel anti-gravitropic mechanism

Circulating microparticles: square the circle

Background: The present review summarizes current knowledge about microparticles (MPs) and provides a systematic overview of last 20 years of research on circulating MPs, with particular focus on their clinical relevance. Results: MPs are a heterogeneous population of cell-derived vesicles, with sizes ranging between 50 and 1000 nm. MPs are capable of transferring peptides, proteins, lipid components, microRNA, mRNA, and DNA from one cell to another without direct cell-to-cell contact. Growing evidence suggests that MPs present in peripheral blood and body fluids contribute to the development and progression of cancer, and are of pathophysiological relevance for autoimmune, inflammatory, infectious, cardiovascular, hematological, and other diseases. MPs have large diagnostic potential as biomarkers; however, due to current technological limitations in purification of MPs and an absence of standardized methods of MP detection, challenges remain in validating the potential of MPs as a non-invasive and early diagnostic platform. Conclusions: Improvements in the effective deciphering of MP molecular signatures will be critical not only for diagnostics, but also for the evaluation of treatment regimens and predicting disease outcomes

Design and baseline characteristics of the finerenone in reducing cardiovascular mortality and morbidity in diabetic kidney disease trial

Background: Among people with diabetes, those with kidney disease have exceptionally high rates of cardiovascular (CV) morbidity and mortality and progression of their underlying kidney disease. Finerenone is a novel, nonsteroidal, selective mineralocorticoid receptor antagonist that has shown to reduce albuminuria in type 2 diabetes (T2D) patients with chronic kidney disease (CKD) while revealing only a low risk of hyperkalemia. However, the effect of finerenone on CV and renal outcomes has not yet been investigated in long-term trials. Patients and Methods: The Finerenone in Reducing CV Mortality and Morbidity in Diabetic Kidney Disease (FIGARO-DKD) trial aims to assess the efficacy and safety of finerenone compared to placebo at reducing clinically important CV and renal outcomes in T2D patients with CKD. FIGARO-DKD is a randomized, double-blind, placebo-controlled, parallel-group, event-driven trial running in 47 countries with an expected duration of approximately 6 years. FIGARO-DKD randomized 7,437 patients with an estimated glomerular filtration rate >= 25 mL/min/1.73 m(2) and albuminuria (urinary albumin-to-creatinine ratio >= 30 to <= 5,000 mg/g). The study has at least 90% power to detect a 20% reduction in the risk of the primary outcome (overall two-sided significance level alpha = 0.05), the composite of time to first occurrence of CV death, nonfatal myocardial infarction, nonfatal stroke, or hospitalization for heart failure. Conclusions: FIGARO-DKD will determine whether an optimally treated cohort of T2D patients with CKD at high risk of CV and renal events will experience cardiorenal benefits with the addition of finerenone to their treatment regimen. Trial Registration: EudraCT number: 2015-000950-39; ClinicalTrials.gov identifier: NCT02545049

Examination of B. subtilis var. niger Spore Killing by Dry Heat Methods

Dry heat microbial reduction is the only NASA approved sterilization method to reduce the microbial bioburden on space-flight hardware prior to launch. Reduction of the microbial bioburden on spacecraft is necessary to meet planetary protection requirements specific for the mission. Microbial bioburden reduction also occurs if a spacecraft enters a planetary atmosphere (e.g., Mars) and is heated due to frictional forces. Temperatures reached during atmospheric entry events (>200 C) are sufficient to damage or destroy flight hardware and also kill microbial spores that reside on the in-bound spacecraft. The goal of this research is to determine the survival rates of bacterial spores when they are subjected to conditions similar to those the spacecraft would encounter (i.e., temperature, pressure, etc.). B. subtilis var. niger spore coupons were exposed to a range of temperatures from 125 C to 200 C in a vacuum oven (at <1 Torr). After the exposures, the spores were removed by sonication, dilutions were made, and the spores were plated using the pour plate method with tryptic soy agar. After 3 days incubation at 32 C, the number of colony-forming units was counted. Lethality rate constants and D-values were calculated at each temperature. The calculated D-values were: 27 minutes (at 125 C), 13 minutes (at 135 C), and <0.1 minutes (at 150 C). The 125 C and 135 C survivor curves appeared as concavedownward curves. The 150 C survivor curve appeared as a straight-line. Due to the prolonged ramp-up time to the exposure conditions, spore killing during the ramp-up resulted in insufficient data to draw curves for exposures at 160 C, 175 C, and 200 C. Exploratory experiments using novel techniques, with short ramp times, for performing high temperature exposures were also examined. Several of these techniques, such as vacuum furnaces, thermal spore exposure vessels, and laser heating of the coupons, will be discussed

Extended temperature range studies for dry heat microbial reduction

This paper will present the lethality data that has been collected at this time and the planned future studies. The results show that rapid ramp-up heating times are critical to obtaining valid lethality data at high temperatures because an extensive number of spores are killed before reaching the target temperature. Exploratory experiments have also been performed using a laser to rapidly heat coupons