ATPase and Multidrug Transport Activities of the Overexpressed Yeast ABC Protein Yor1p

The Saccharomyces cerevisiae genome encodes 15 full-size ATP binding cassette transporters (ABC), of which PDR5, SNQ2, and YOR1 are known to be regulated by the transcription factors Pdr1p and Pdr3p (pleiotropic drug resistance). We have identified two new ABC transporter-encoding genes, PDR10 and PDR15, which were up-regulated by the PDR1-3 mutation. These genes, as well as four other ABC transporter-encoding genes, were deleted in order to study the properties of Yor1p. The PDR1-3 gain-of-function mutant was then used to overproduce Yor1p up to 10% of the total plasma membrane proteins. Overexpressed Yor1p was photolabeled by [gamma-32P]2', 3'-O-(2,4,6-trinitrophenyl)-8-azido-ATP (K0.5 = 45 microM) and inhibited by ATP (KD = 0.3 mM) in plasma membranes. Solubilization and partial purification on sucrose gradient allowed to detect significant Yor1p ATP hydrolysis activity (approximately 100 nmol of Pi.min-1.mg-1). This activity was phospholipid-dependent and sensitive to low concentrations of vanadate (I50 = 0.3 microM) and oligomycin (I50 = 8.5 microg/ml). In vivo, we observed a correlation between the amount of Yor1p in the plasma membrane and the level of resistance to oligomycin. We also demonstrated that Yor1p drives an energy-dependent, proton uncoupler-insensitive, cellular extrusion of rhodamine B. Furthermore, cells lacking both Yor1p and Pdr5p (but not Snq2p) showed increased accumulation of the fluorescent derivative of 1-myristoyl-2-[6-(NBD)aminocaproyl]phosphatidylethanolamine. Despite their different topologies, both Yor1p and Pdr5p mediated the ATP-dependent translocation of similar drugs and phospholipids across the yeast cell membrane. Both ABC transporters exhibit ATP hydrolysis in vitro, but Pdr5p ATPase activity is about 15 times higher than that of Yor1p, which may indicate mechanistic or regulatory differences between the two enzymes

Differences in Thrombotic Risk Factors in Black and White Women with Adverse Pregnancy Outcome

Black women have an increased risk of adverse pregnancy outcomes and the characteristics of thrombotic risk factors in this population are unknown. The objective of this study was to examine the racial differences in thrombotic risk factors among women with adverse pregnancy outcomes

Characteristics of abdominal vein thrombosis in children and adults

The demographic and clinical characteristics of adults and children with lower extremity deep-vein thrombosis and/or pulmonary embolism (LE DVT/PE) may differ from those with abdominal vein thrombosis (abdominal VT). Abdominal VT can be a presenting sign of an underlying prothrombotic state, and its presence in the setting of known disease might have prognostic implications different from LE DVT/PE. This study describes clinical presentations of abdominal VT compared to LE DVT/PE in adults and children. We analysed prospec-tively-collected data from consecutive consenting patients enrolled in one of seven Centers for Disease Control and Prevention (CDC) funded Thrombosis and Hemostasis Network Centers from August 2003 to April 2011 to compare the demographic and clinical characteristics of adults and children with abdominal VT. Both adults and children with abdominal VT tended to be younger and have a lower body mass index (BMI) than those with LE DVT/PE. Of patients with abdominal V T, children were more likely to have inferior vena cava (IVC) thrombosis than adults. For adults with venous thromboembolism (VTE), relatively more women had abdominal VT than LE DVT/PE, while the proportions with LE DVT/PE and abdominal VT by sex were similar in children. Children with abdominal VT were more likely to have diagnosed inherited thrombophilia, while trauma was more common in children with LE DVT/PE. In conclusion, both children and adults with abdominal VT were younger with a lower BMI than those with LE DVT/ PE. Significant differences exist between children and adults in respect to abdominal VT compared to LE DVT/PE

Whole blood gene expression profiles distinguish clinical phenotypes of venous thromboembolism

Recurrent venous thromboembolism (VTE) occurs infrequently following a provoked event but occurs in up to 30% of individuals following an initial unprovoked event. There is limited understanding of the biological mechanisms that predispose patients to recurrent VTE

Overview of Sickle Cell Disease Activities at the Centers for Disease Control and Prevention

Presented on June 23, 2011 from 2:00 pm to 5:30 pm, Klaus 1116E on the Georgia Tech Campus.Runtime: 18:04 minutesThe Centers for Disease Control and Prevention’s research and programmatic activities for sickle cell disease are focused on four main areas: Public Health Surveillance, Epidemiological and Laboratory Research, Prevention, Education, and Awareness, and Global Technical Assistance. With respect to public health surveillance, CDC in collaboration with the National Institutes of Health and seven state health departments (GA, NC, CA, MI, PA, NY and FL) is in the second year of a pilot project to develop the first ever public health surveillance system to assess the burden and monitor the health of persons with hemoglobinopathies including sickle cell disease and thalassemia. Also in the area of public health surveillance, in 2011, CDC in also embarking on a pilot expansion of a blood safety monitoring program to assess the adverse health outcomes of person with blood disorders including sickle cell disease who are therapeutically transfused. With respect to epidemiological and laboratory research CDC researchers lead and/or collaborate with other investigators examining the a host of outcomes among the sickle cell disease population including maternal and child health outcomes, asthma, mortality, acute chest syndrome and others. Additionally CDC is stimulating awareness, collaboration and education through sponsorship of events, scientific meetings, webinars, and workgroups. Finally, CDC is providing technical assistance to several international partners to assist those partners with developing laboratory capacity for the establishment of newborn screening and developing data collection and surveillance systems to monitor and evaluate the impact of public health and health programs for sickle cell disease in those countries

Different heating techniques for oat lipase stabilization: Grain quality and storability

Incorporation of oats in the human diet has received great attention for its role in the lowering of blood plasma cholesterol levels; however, problems associated with postprocessing storage continue to limit its use. The action of lipases is believed to be the main problem. When lipase comes in contact with a lipid substrate after milling, it results in deteriorative effects such as oxidation that cause the development of off flavors and odors. The current industrial heat treatment uses convective heat and live steam to inactivate oat lipases. Alternative heat applications, including microwave (MW) energy, infrared (IR) processing, conventional oven heating (COH) and a simulated industrial heat treatment (IHT) were administered to samples of commercial whole oat kernels. Results were analyzed using the repeated measures procedure of the Statistical Analysis System (SAS) program (v6.12). Analysis of variance (ANOVA) was performed by following the General Linear Model (GLM) procedure for SAS. Principle component factor analysis was also performed. Treatment differences were determined using the LSD test and were reported to be significant when P \u3c 0.05. Significantly greater P \u3c 0.05. lipase activity was shown during oat storage in the control (CTRL) oats (with no heat treatment) after 3 and 6 months than for heat treated oats. IR treatment contained the least amount of activity after 3 months, but was not significantly different than IHT treated oats. POV was significantly higher P \u3c 0.05. for IF, MW and IHT heat treatments initially than for the other treatments, but decreased more for the IR and MW treatments over time. The COH, CTRL and IHT treatments exhibited a sharper reduction in POV after 3 months storage than for the IR and MW samples. FA levels were significantly different P \u3c 0.05. for IR treatments than for other heat applications, where an increasing trend was observed for C14:0 and C16:0/16:1 saturated and monounsaturated fatty acids; and a decreasing trend for C18:2 and C18:3 unsaturated fatty acids. SPME headspace analysis found that hexanal, the major volatile formed, was observed initially for IR treated oats only. When detected, it was present only at trace levels in (W-O) samples of the CTRL, COH, MW and IHT treatments. Although not significant, (G-O) samples produced higher hexanal volatiles than (W-O) samples of the same heat treatment, and were highest for the CTRL and IR treatments. IR treated (W-O) samples were the exception, however, where hexanal in (W-O) was higher than for (G-O) during the first 2 months, before decreasing to much lower levels for the remaining storage period. (Abstract shortened by UMI.

Predictive accuracy of 29-comorbidity index for in-hospital deaths in US adult hospitalizations with a diagnosis of venous thromboembolism.

BACKGROUND: Venous thromboembolism (VTE), comprising deep vein thrombosis (DVT) and pulmonary embolism (PE), is a significant source of mortality and morbidity worldwide. By analyzing data of the 2010 Nationwide Inpatient Sample from the Agency for Healthcare Research and Quality (AHRQ), we evaluated the predictive accuracy of the AHRQ's 29-comorbidity index with in-hospital death among US adult hospitalizations with a diagnosis of VTE. METHODS: We assessed the case-fatality and prevalence of comorbidities among a sample of 153,518 adult hospitalizations with a diagnosis of VTE that comprised 87,605 DVTs and 65,913 PEs (with and without DVT). We estimated adjusted odds ratios and 95% confidence intervals with multivariable logistic regression models by using comorbidities as predictors and status of in-hospital death as an outcome variable. We assessed the c-statistics for the predictive accuracy of the logistic regression models. RESULTS: In 2010, approximately 41,944 in-hospital deaths (20,212 with DVT and 21,732 with PE) occurred among 770,137 hospitalizations with a diagnosis of VTE. When compared separately to hospitalizations with VTE, DVT, or PE that had no corresponding comorbidities, congestive heart failure, chronic pulmonary disease, coagulopathy, liver disease, lymphoma, fluid and electrolyte disorders, metastatic cancer, other neurological disorders, peripheral vascular disorders, pulmonary circulation disorders, renal failure, solid tumor without metastasis, and weight loss were positively and independently associated with 10%-125% increased likelihoods of in-hospital death. The c-statistic values ranged from 0.776 to 0.802. CONCLUSION: The results of this study indicated that comorbidity was associated independently with risk of death among hospitalizations with VTE and among hospitalizations with DVT or PE. The AHRQ 29-comorbidity index provides acceptable to excellent predictive accuracy for in-hospital deaths among adult hospitalizations with VTE and among those with DVT or PE