Study of biochemical changes after plateletpheresis in healthy male donors

BACKGROUND: There is relatively little information about endogenous biochemical changes in a response to plateletpheresis in healthy donors. We aimed to investigate the changes in different biochemical parameters including glycemic status, insulin resistance, iron status, lipid profile and inflammatory markers after plateletpheresis in healthy male donors with normal glycemic status. METHODS: In this study we enrolled 10 male subjects. The glycemic status in all subjects was assessed using an oral glucose tolerance test pre- and post-plateletpheresis at different time intervals (1, 8 and 22 days). Different biochemical parameters including glucose, HbA1c, insulin, lipids, uric acid, transferrin, ferritin, C-reactive protein and insulin resistance were measured. Repeated ANOVA was utilized for the purpose of statistical comparison of means between different days. RESULTS: Fasting glucose, transferrin, cholesterol, triglycerides, HDL-C, and LDL-C were significantly altered (-3.9%, p<0.05; -2.7%, p<0.05; -3.9%, p<0.05; 23.9%, p<0.05; -5.5%, p<0.01; and -9.2%, p<0.05 respectively) at day 1 following plateletpheresis. There was a gradual reduction in HbA1c and ferritin levels during the time-course of the study, and by day 22, both were significantly lower (-2.0%, p<0.01; -18.1%, p<0.05 respectively) when compared to the pre-plateletpheresis levels. CONCLUSIONS: Post-plateletpheresis, several biochemical parameters may change significantly in healthy donors. The changes were particularly evident one and 22 days after donation. The potential effects of plateletpheresis need to be considered when interpreting biochemical tests

A role for the mevalonate pathway in early plant symbiotic signaling

Rhizobia and arbuscular mycorrhizal fungi produce signals that are perceived by host legume receptors at the plasma membrane and trigger sustained oscillations of the nuclear and perinuclear Ca(2+) concentration (Ca(2+) spiking), which in turn leads to gene expression and downstream symbiotic responses. The activation of Ca(2+) spiking requires the plasma membrane-localized receptor-like kinase Does not Make Infections 2 (DMI2) as well as the nuclear cation channel DMI1. A key enzyme regulating the mevalonate (MVA) pathway, 3-Hydroxy-3-Methylglutaryl CoA Reductase 1 (HMGR1), interacts with DMI2 and is required for the legume-rhizobium symbiosis. Here, we show that HMGR1 is required to initiate Ca(2+) spiking and symbiotic gene expression in Medicago truncatula roots in response to rhizobial and arbuscular mycorrhizal fungal signals. Furthermore, MVA, the direct product of HMGR1 activity, is sufficient to induce nuclear-associated Ca(2+) spiking and symbiotic gene expression in both wild-type plants and dmi2 mutants, but interestingly not in dmi1 mutants. Finally, MVA induced Ca(2+) spiking in Human Embryonic Kidney 293 cells expressing DMI1. This demonstrates that the nuclear cation channel DMI1 is sufficient to support MVA-induced Ca(2+) spiking in this heterologous system

Engineering and Two-Stage Evolution of a Lignocellulosic Hydrolysate-Tolerant <i>Saccharomyces cerevisiae</i> Strain for Anaerobic Fermentation of Xylose from AFEX Pretreated Corn Stover

<div><p>The inability of the yeast <i>Saccharomyces cerevisiae</i> to ferment xylose effectively under anaerobic conditions is a major barrier to economical production of lignocellulosic biofuels. Although genetic approaches have enabled engineering of <i>S. cerevisiae</i> to convert xylose efficiently into ethanol in defined lab medium, few strains are able to ferment xylose from lignocellulosic hydrolysates in the absence of oxygen. This limited xylose conversion is believed to result from small molecules generated during biomass pretreatment and hydrolysis, which induce cellular stress and impair metabolism. Here, we describe the development of a xylose-fermenting <i>S. cerevisiae</i> strain with tolerance to a range of pretreated and hydrolyzed lignocellulose, including Ammonia Fiber Expansion (AFEX)-pretreated corn stover hydrolysate (ACSH). We genetically engineered a hydrolysate-resistant yeast strain with bacterial xylose isomerase and then applied two separate stages of aerobic and anaerobic directed evolution. The emergent <i>S. cerevisiae</i> strain rapidly converted xylose from lab medium and ACSH to ethanol under strict anaerobic conditions. Metabolomic, genetic and biochemical analyses suggested that a missense mutation in <i>GRE3</i>, which was acquired during the anaerobic evolution, contributed toward improved xylose conversion by reducing intracellular production of xylitol, an inhibitor of xylose isomerase. These results validate our combinatorial approach, which utilized phenotypic strain selection, rational engineering and directed evolution for the generation of a robust <i>S. cerevisiae</i> strain with the ability to ferment xylose anaerobically from ACSH.</p></div