Cross-talk between cognate and noncognate RpoE sigma factors and Zn(2+)-binding anti-sigma factors regulates photooxidative stress response in Azospirillum brasilense.

PubMed

Gupta, Namrata; Gupta, Ankush; Kumar, Santosh; Mishra, Rajeev; Singh, Chhaya; Tripathi, Anil Kumar

2014-01-01

Azospirillum brasilense harbors two redox-sensitive Zinc-binding anti-sigma (ZAS) factors (ChrR1 and ChrR2), which negatively regulate the activity of their cognate extra-cytoplasmic function (ECF) σ factors (RpoE1 and RpoE2) by occluding their binding to the core enzyme. Both pairs of RpoE-ChrR control responses to photooxidative stress. The aim of this study was to investigate whether the two RpoE-ChrR pairs cross-talk while responding to the stress. In silico analysis showed a high sequence similarity between ChrR1 and ChrR2 proteins, but differences in redox sensitivity. Using in silico and in vitro methods of protein-protein interaction, we have shown that both ChrR1 and ChrR2 proteins physically bind to their noncognate RpoE proteins. Restoration of the phenotypes of chrR1::Tn5 and chrR2::Km mutants related to carotenoid biosynthesis and photooxidative stress tolerance by expressing chrR1 or chrR2 provided in vivo evidence for the cross-talk. In addition, up- or down-regulation of several identical proteins by expressing chrR1 or chrR2 in the chrR1::Tn5 mutant provided another in vivo evidence for the cross-talk. Although multiple redox-sensitive ZAS anti-σ factors occur in some Gram-positive bacteria, no cross-talk is reported among them. We report here, for the first time, that the two ZAS anti-σ factors of A. brasilense also interact with their noncognate σ factors and affect gene expression. The two redox-sensitive ZAS anti-σ factors in A. brasilense may interact with their cognate as well as noncognate ECF σ factors to play an important role in redox homeostasis by facilitating recovery from the oxidative stress.

Extracellular Fluid/Intracellular Fluid Volume Ratio as a Novel Risk Indicator for All-Cause Mortality and Cardiovascular Disease in Hemodialysis Patients

PubMed Central

Kim, Eun-Jung; Choi, Myung-Jin; Lee, Jeoung-Hwan; Oh, Ji-Eun; Seo, Jang-Won; Lee, Young-Ki; Yoon, Jong-Woo; Kim, Hyung-Jik; Noh, Jung-Woo

2017-01-01

Background In hemodialysis patients, fluid overload and malnutrition are accompanied by extracellular fluid (ECF) expansion and intracellular fluid (ICF) depletion, respectively. We investigated the relationship between ECF/ICF ratio (as an integrated marker reflecting both fluid overload and malnutrition) and survival and cardiovascular disease (CVD) in the context of malnutrition-inflammation-arteriosclerosis (MIA) complex. Methods Seventy-seven patients from a single hemodialysis unit were prospectively enrolled. The ECF/ICF volume was measured by segmental multi-frequency bioimpedance analysis. MIA and volume status were measured by serum albumin, C-reactive protein (CRP), pulse wave velocity (PWV) and plasma B-type natriuretic peptide (BNP), respectively. Results The mean ECF/ICF ratio was 0.56±0.06 and the cut-off value for maximum discrimination of survival was 0.57. Compared with the low ECF/ICF group, the high ECF/ICF group (ratio≥0.57, 42%) had higher all-cause mortality, CVD, CRP, PWV, and BNP, but lower serum albumin. During the 5-year follow-up, 24 all-cause mortality and 38 CVD occurred (18 and 24, respectively, in the high ECF/ICF group versus 6 and 14 respectively in the low ECF/ICF group, P<0.001). In the adjusted Cox analysis, the ECF/ICF ratio nullifies the effects of the MIA and volume status on survival and CVD and was an independent predictor of all-cause mortality and CVD: hazard ratio (95% confidence interval); 1.12 (1.01–1.25) and 1.09 (1.01–1.18) for a 0.01 increase in the ECF/ICF ratio. The degree of malnutrition (albumin), inflammation (CRP), arteriosclerosis (PWV), and fluid overload (BNP) were correlated well with the ECF/ICF ratio. Conclusions Hemodialysis patients with high ECF/ICF ratio are not only fluid overloaded, but malnourished and have stiff artery with more inflammation. The ECF/ICF ratio is highly related to the MIA complex, and is a major risk indicator for all-cause mortality and CVD. PMID:28099511

Enterocutaneous Fistulae: Etiology, Treatment, and Outcome – A Study from South India

PubMed Central

Kumar, Prakash; Maroju, Nanda K.; Kate, Vikram

2011-01-01

Background/Aim: Enterocutaneous fistula (ECF) is a difficult condition managed in the surgical wards and is associated with significant morbidity and mortality. Sepsis, malnutrition, and electrolyte abnormality is the classical triad of complications of ECF. Sepsis with malnutrition is the leading cause of death in cases of ECF. Although it is a common condition, no recent report in literature on the profile of patients with ECF has been documented from the southern part of India. Materials and Methods: All consecutive patients who developed or presented with ECF during the study period were included in the study. The etiology, anatomic distribution, fistula output, clinical course, complications, predictive factors for spontaneous closure, and outcomes for patients with ECF were studied. Results: A total of 41 patients were included in this prospective observational study, of which 34 were males and 7 were females. About 95% of ECF were postoperative. Ileum was found to be the most common site of ECF. Also, 49% of fistulas were high output and 51% were low output. Serum albumin levels correlated significantly with fistula healing and mortality. Surgical intervention was required in 41% of patients. Conclusion: Most of the ECF are encountered in the postoperative period. Serum albumin levels can predict fistula healing and mortality. Conservative management should be the first line of treatment. Mortality in patients with ECF continues to be significant and is commonly related to malnutrition and sepsis. PMID:22064337

Immune parameters to p67C antigen adjuvanted with ISA206VG correlate with protection against East Coast fever.

PubMed

Lacasta, Anna; Mwalimu, Stephen; Kibwana, Elisabeth; Saya, Rosemary; Awino, Elias; Njoroge, Thomas; Poole, Jane; Ndiwa, Nicholas; Pelle, Roger; Nene, Vishvanath; Steinaa, Lucilla

2018-03-07

East Coast fever (ECF) is a lymphoproliferative disease caused by the tick-transmitted protozoan parasite Theileria parva. ECF is one of the most serious cattle tick-borne diseases in Sub-Saharan Africa. We have previously demonstrated that three doses of the C-terminal part of the sporozoite protein p67 (p67C) adjuvanted with ISA206VG confers partial protection against ECF at a herd level. We have tested the efficacy of two doses of this experimental vaccine, as reducing the vaccination regimen would facilitate its deployment in the field. We reconfirm that three antigen doses gave a significant level of protection to severe disease (46%, ECF score < 6) when compared with the control group, while two doses did not (23%). Animals receiving three doses of p67C developed higher antibody titers and CD4 + T-cell proliferation indices, than those which received two doses. A new panel of immune parameters were tested in order to identify factors correlating with protection: CD4 + proliferation index, total IgG, IgG1, IgG2 and IgM half maximal titers and neutralization capacity of the sera with and without complement. We show that some of the cellular and humoral immune responses provide preliminary correlates of protection. Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.

[Nutritional support and risk factors of appearance of enterocutaneous fistulas].

PubMed

Llop, J M; Cobo, S; Padullés, A; Farran, L; Jódar, R; Badia, M B

2012-01-01

Among the different factors described, nutritional support has been associated to prevention and management of enterocutaneous fistulae (ECF). To assess the influence that the parameters related to nutritional, clinical status, and surgical variables have on the occurrence of ECF. An observational case/control retrospective study was performed on patients admitted to the General and Digestive Surgery Department. The parameters analyzed were: diagnosis, body mass index (BMI), pathologic personal history, number of surgical interventions (SI) and complications (previous infection, bleeding, and ischemia). In patients with SI, we analyzed: number and type of SI, time until onset of nutritional support, and type of nutritional support. We performed a multiple logistic uni- and multivariate regression analysis by using the SPSSv.19.0 software. The primary diagnoses related to the occurrence of ECF were pancreatic pathology (OR = 5.346) and inflammatory bowel disease (IBD) (OR = 9.329). The surgical variables associated to higher prevalence of ECF emergency SI (OR = 5.79) and multiple SI (OR = 4.52). Regarding the nutritional variables, the late onset of nutrition (more than three days after SI) was associated to the occurrence of ECF (OR = 3.82). In surgical patients, early nutritional support , independently of the route of administration, decreases the occurrence of fistulae. Pancreatic pathology, IBD, emergency SI, and multiple SI were associated to higher prevalence of ECF. The variable hyponutrition appears as a risk factor that should be confirmed in further studies.

Efficacy and safety of mitomycin C as an agent to treat corneal scarring in horses using an in vitro model

PubMed Central

Buss, Dylan G.; Sharma, Ajay; Giuliano, Elizabeth A.; Mohan, Rajiv R.

2010-01-01

Objective Mitomycin C (MMC) is used clinically to treat corneal scarring in human patients. We investigated the safety and efficacy of MMC to treat corneal scarring in horses by examining its effects at the early and late stages of disease using an in-vitro model. Procedure An in-vitro model of equine corneal fibroblast (ECF) developed was used. The equine corneal fibroblast or myofibroblast cultures were produced by growing primary ECF in the presence or absence of transforming growth factor beta-1 (TGFβ1) under serum-free conditions. The MMC dose for the equine cornea was defined with dose-dependent trypan blue exclusion and MTT [(3-4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assays after applying MMC to the cultures once for 2 minutes. The efficacy of MMC to control corneal scarring in horses was determined by measuring mRNA and protein expression of corneal scarring markers (α-smooth muscle actin and F-actin) with western blotting, immunocytochemistry and/or quantitative real-time polymerase chain reactions. Results A single 2 minutes treatment of 0.02% or less MMC did not alter ECF phenotype, viability, or cellular proliferation whereas 0.05% or higher MMC doses showed mild-to-moderate cellular toxicity. The TGFβ1 at 1ng/ml showed significant myofibroblast formation in ECF under serum-free conditions. A single 2 minute, 0.02% MMC treatment 24 hours (early) after TGFβ1 stimulation significantly reduced conversion of ECF to myofibroblasts, however, a single 0.02% MMC treatment 11 days after TGFβ1 stimulation showed moderate myofibroblast inhibition. Conclusions That MMC safely and effectively reduced scarring in ECF by reducing the degree of transdifferentiation of corneal fibroblasts to myofibroblasts in vitro. Further clinical in-vivo investigations are warranted using MMC in horses. PMID:20618797

Nutritional Management in Enterocutaneous Fistula. What is the evidence?

PubMed Central

BADRASAWI, Manal; SHAHAR, Suzana; SAGAP, Ismail

2015-01-01

The management of Enterocutaneous fistula (ECF) is challenging. It remains associated with morbidity and mortality, despite advancements in medical and surgical therapies. Early nutritional support using parenteral, enteral or fystuloclysis routs is essential to reverse catabolism and replace nutrients, fluid and electrolyte losses. This study aims to review the current literature on the management of ECF. Fistulae classifications have an impact on the calories and protein requirements. Early nutritional support with parenteral, enteral nutrition or fistuloclysis played a significant role in the management outcome. Published literature on the nutritional management of ECF is mostly retrospective and lacks experimental design. Prospective studies do not investigate nutritional assessment or management experimentally. Individualising the nutritional management protocol was recommended due to the absence of management guidelines for ECF patients. PMID:26715903

In vitro Antiproliferative Effect of Earthworm Coelomic Fluid of Eudrilus Eugeniae, Eisenia Foetida, and Perionyx Excavatus on Squamous Cell Carcinoma-9 Cell Line: A Pilot Study.

PubMed

Augustine, Dominic; Rao, Roopa S; Anbu, Jayaraman; Chidambara Murthy, K N

2017-12-01

The earthworm coelomic fluid (ECF) has shown proven antiproliferative effect against breast, liver, gastrointestinal, and brain cancer, but it is least explored in oral cancer. The present in vitro study is an attempt to investigate the antiproliferative activity of ECF on oral cancer cell line squamous cell carcinoma (SCC)-9. ECF was collected from the species Eudrilus eugeniae (EE), Eisenia foetida (EF), and Perionyx excavatus (PE) stored at -80°C. Percentage inhibition of ECF on squamous cell carcinoma-9 cells in vitro was recorded at 24 h. Protein estimation was done using Bradford protein assay validated by the biuret method. Cytotoxicity was tested at 2.5, 5, 10, 20, 40, and 80 μg/ml concentrations by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay in SCC-9 cells in vitro . GraphPad Prism 7.0 software was used to calculate the inhibitory concentration (IC 50 ). Chi-square test was used to analyze the difference between samples. The test samples EE, EF, and PE inhibited the growth of SCC-9 cells significantly in a dose-dependent manner, and the IC 50 values were found to be 4.6, 44.69, and 5.27 μg/ml, respectively. The antiproliferative effect was found to be variable among the three earthworm species with EE showing the most promising effect followed by PE and EF. Establishing the antiproliferative effect of ECF on oral cancer cells could be an initial step toward drug development and future anticancer research. The preliminary investigation has shown that ECF has a promising antiproliferative effect on oral cancer cells in vitro . The present pilot study evaluated the in vitro antiproliferative effect of earthworm coelomic fluid (ECF) of Eudrilus eugeniae (EE), Eisenia foetida (EF), and Perionyx excavatus (PE) on squamous cell carcinoma-9 cell line. The ECF inhibitory activity was promising at inhibitory concentration values of 4.6, 44.69, and 5.27 μg/ml, respectively. Further studies pertaining to antiproliferative mechanism of EE, EF, and PE have been planned. Abbreviations Used: ECF: Earthworm coelomic fluid, EE: Eudrilus eugeniae , EF: Eisenia foetida , PE: Perionyx excavatus , MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, SCC: Squamous cell carcinoma, BSA: Bovine serum albumin, PBS: Phosphored buffered saline, ATCC: American Type Culture Collection.

Definitive surgical closure of enterocutaneous fistula: outcome and factors predictive of increased postoperative morbidity.

PubMed

Ravindran, P; Ansari, N; Young, C J; Solomon, M J

2014-03-01

Enterocutaneous fistula (ECF) presents a complex management problem with significant mortality and morbidity. The aim of this study was to assess the outcome of patients undergoing surgical cure for ECF and to predict factors that might relate to increased postoperative morbidity. Medical records of all patients who underwent definitive surgery for cure of an ECF within our colorectal surgery unit between 2000 and 2010 were reviewed. Forty-one patients (18 male) were identified, in whom 44 definitive procedures were performed. The median age was 54 (17-81) years. The median postoperative length of stay in hospital was 14 (2-213) days. Half (50%) of the ECFs occurred as a postoperative complication followed by spontaneous fistulation in Crohn's disease (36%). The interval to definitive surgery was influenced by the aetiology of the fistula. The median time to surgery after formation of postoperative fistula was 240 days (7.9 months). There was no 30-day postoperative mortality. There were two (4.5%) recurrences at 3 months. Thirty-eight (86%) patients suffered postoperative morbidity as defined by the Clavien-Dindo classification. High-grade morbidity occurred in 32% of patients. On univariate analysis, factors identified as being significantly associated with high-grade morbidity included a fistula output of > 500 ml/day (P = 0.004) in patients with postoperative ECF, malnutrition at presentation (P = 0.04) and a serum albumin value of < 30 g/l (P = 0.02) in patients with spontaneous ECF due to Crohn's disease. The majority of persistent complex ECFs can be cured surgically with low mortality and recurrence in a multidisciplinary setting. Postoperative morbidity, however, remains a significant burden. Colorectal Disease © 2013 The Association of Coloproctology of Great Britain and Ireland.

SigCH, an extracytoplasmic function sigma factor of Porphyromonas gingivalis regulates the expression of cdhR and hmuYR.

PubMed

Ota, Koki; Kikuchi, Yuichiro; Imamura, Kentaro; Kita, Daichi; Yoshikawa, Kouki; Saito, Atsushi; Ishihara, Kazuyuki

2017-02-01

Extracytoplasmic function (ECF) sigma factors play an important role in the bacterial response to various environmental stresses. Porphyromonas gingivalis, a prominent etiological agent in human periodontitis, possesses six putative ECF sigma factors. So far, information is limited on the ECF sigma factor, PGN_0319. The aim of this study was to investigate the role of PGN_0319 (SigCH) of P. gingivalis, focusing on the regulation of hmuY and hmuR, which encode outer-membrane proteins involved in hemin utilization, and cdhR, a transcriptional regulator of hmuYR. First, we evaluated the gene expression profile of the sigCH mutant by DNA microarray. Among the genes with altered expression levels, those involved in hemin utilization were downregulated in the sigCH mutant. To verify the microarray data, quantitative reverse transcription PCR analysis was performed. The RNA samples used were obtained from bacterial cells grown to early-log phase, in which sigCH expression in the wild type was significantly higher than that in mid-log and late-log phases. The expression levels of hmuY, hmuR, and cdhR were significantly decreased in the sigCH mutant compared to wild type. Transcription of these genes was restored in a sigCH complemented strain. Compared to the wild type, the sigCH mutant showed reduced growth in log phase under hemin-limiting conditions. Electrophoretic mobility shift assays showed that recombinant SigCH protein bound to the promoter region of hmuY and cdhR. These results suggest that SigCH plays an important role in the early growth of P. gingivalis, and directly regulates cdhR and hmuYR, thereby playing a potential role in the mechanisms of hemin utilization by P. gingivalis. Copyright © 2016 Elsevier Ltd. All rights reserved.

Sample dilution and bacterial community composition influence empirical leucine-to-carbon conversion factors in surface waters of the world's oceans.

PubMed

Teira, Eva; Hernando-Morales, Víctor; Cornejo-Castillo, Francisco M; Alonso-Sáez, Laura; Sarmento, Hugo; Valencia-Vila, Joaquín; Serrano Catalá, Teresa; Hernández-Ruiz, Marta; Varela, Marta M; Ferrera, Isabel; Gutiérrez Morán, Xosé Anxelu; Gasol, Josep M

2015-12-01

The transformation of leucine incorporation rates to prokaryotic carbon production rates requires the use of either theoretical or empirically determined conversion factors. Empirical leucine-to-carbon conversion factors (eCFs) vary widely across environments, and little is known about their potential controlling factors. We conducted 10 surface seawater manipulation experiments across the world's oceans, where the growth of the natural prokaryotic assemblages was promoted by filtration (i.e., removal of grazers [F treatment]) or filtration combined with dilution (i.e., also relieving resource competition [FD treatment]). The impact of sunlight exposure was also evaluated in the FD treatments, and we did not find a significant effect on the eCFs. The eCFs varied from 0.09 to 1.47 kg C mol Leu(-1) and were significantly lower in the FD than in the F samples. Also, changes in bacterial community composition during the incubations, as assessed by automated ribosomal intergenic spacer analysis (ARISA), were more pronounced in the FD than in the F treatments, compared to unmanipulated controls. Thus, we discourage the common procedure of diluting samples (in addition to filtration) for eCF determination. The eCFs in the filtered treatment were negatively correlated with the initial chlorophyll a concentration, picocyanobacterial abundance (mostly Prochlorococcus), and the percentage of heterotrophic prokaryotes with high nucleic acid content (%HNA). The latter two variables explained 80% of the eCF variability in the F treatment, supporting the view that both Prochlorococcus and HNA prokaryotes incorporate leucine in substantial amounts, although this results in relatively low carbon production rates in the oligotrophic ocean. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Sample Dilution and Bacterial Community Composition Influence Empirical Leucine-to-Carbon Conversion Factors in Surface Waters of the World's Oceans

PubMed Central

Hernando-Morales, Víctor; Cornejo-Castillo, Francisco M.; Alonso-Sáez, Laura; Sarmento, Hugo; Valencia-Vila, Joaquín; Serrano Catalá, Teresa; Hernández-Ruiz, Marta; Varela, Marta M.; Ferrera, Isabel; Gutiérrez Morán, Xosé Anxelu; Gasol, Josep M.

2015-01-01

The transformation of leucine incorporation rates to prokaryotic carbon production rates requires the use of either theoretical or empirically determined conversion factors. Empirical leucine-to-carbon conversion factors (eCFs) vary widely across environments, and little is known about their potential controlling factors. We conducted 10 surface seawater manipulation experiments across the world's oceans, where the growth of the natural prokaryotic assemblages was promoted by filtration (i.e., removal of grazers [F treatment]) or filtration combined with dilution (i.e., also relieving resource competition [FD treatment]). The impact of sunlight exposure was also evaluated in the FD treatments, and we did not find a significant effect on the eCFs. The eCFs varied from 0.09 to 1.47 kg C mol Leu−1 and were significantly lower in the FD than in the F samples. Also, changes in bacterial community composition during the incubations, as assessed by automated ribosomal intergenic spacer analysis (ARISA), were more pronounced in the FD than in the F treatments, compared to unmanipulated controls. Thus, we discourage the common procedure of diluting samples (in addition to filtration) for eCF determination. The eCFs in the filtered treatment were negatively correlated with the initial chlorophyll a concentration, picocyanobacterial abundance (mostly Prochlorococcus), and the percentage of heterotrophic prokaryotes with high nucleic acid content (%HNA). The latter two variables explained 80% of the eCF variability in the F treatment, supporting the view that both Prochlorococcus and HNA prokaryotes incorporate leucine in substantial amounts, although this results in relatively low carbon production rates in the oligotrophic ocean. PMID:26407885

The Bartonella quintana Extracytoplasmic Function Sigma Factor RpoE Has a Role in Bacterial Adaptation to the Arthropod Vector Environment

PubMed Central

Abromaitis, Stephanie

2013-01-01

Bartonella quintana is a vector-borne bacterial pathogen that causes fatal disease in humans. During the infectious cycle, B. quintana transitions from the hemin-restricted human bloodstream to the hemin-rich body louse vector. Because extracytoplasmic function (ECF) sigma factors often regulate adaptation to environmental changes, we hypothesized that a previously unstudied B. quintana ECF sigma factor, RpoE, is involved in the transition from the human host to the body louse vector. The genomic context of B. quintana rpoE identified it as a member of the ECF15 family of sigma factors found only in alphaproteobacteria. ECF15 sigma factors are believed to be the master regulators of the general stress response in alphaproteobacteria. In this study, we examined the B. quintana RpoE response to two stressors that are encountered in the body louse vector environment, a decreased temperature and an increased hemin concentration. We determined that the expression of rpoE is significantly upregulated at the body louse (28°C) versus the human host (37°C) temperature. rpoE expression also was upregulated when B. quintana was exposed to high hemin concentrations. In vitro and in vivo analyses demonstrated that RpoE function is regulated by a mechanism involving the anti-sigma factor NepR and the response regulator PhyR. The ΔrpoE ΔnepR mutant strain of B. quintana established that RpoE-mediated transcription is important in mediating the tolerance of B. quintana to high hemin concentrations. We present the first analysis of an ECF15 sigma factor in a vector-borne human pathogen and conclude that RpoE has a role in the adaptation of B. quintana to the hemin-rich arthropod vector environment. PMID:23564167

Enterocutaneous fistula in patients with peritoneal malignancy following cytoreductive surgery and hyperthermic intraperitoneal chemotherapy: Incidence, management and outcomes.

PubMed

Valle, Sarah J; Alzahrani, Nayef; Alzahrani, Saleh; Traiki, Thamer Bin; Liauw, Winston; Morris, David L

2016-09-01

Cytoreductive surgery and hyperthermic intraperitoneal chemotherapy (CRS/HIPEC) is an effective treatment for peritoneal carcinomatosis (PC) from multiple origins, however is associated with increased complications compared to conventional gastrointestinal surgery. The aetiology of enterocutaneous fistulas (ECF) in most cases is a result of various contributing factors and therefore remains a major clinical problem, occurring in 4-34% of patients post-CRS. The aim of this study was to analyze the incidence and outcome of ECF following CRS/HIPEC. From April 1999 to September 2015, 53 patients of 918 CRS/HIPEC procedures developed an ECF. Patient, operative and postoperative data were retrospectively analyzed to determine aetiology, classification outcome and possible contributing factors were reviewed on univariate and multivariate analysis. We report a 5.8% ECF rate, diagnosed at a median of 13 days. The mortality rate was 5.7% and other morbidity was significantly increased (p = 0.0001). Twenty-five (47.2%), 8 (15.1%) and 20 patients (37.7%) had low, moderate and high output ECF respectively. Patients that had a CC2 cytoreduction, abdominal VAC or smoked had a higher risk of fistula (p = 0.004, p < 0.0001, p = 0.008). Spontaneous closure was achieved in 49.2% with conservative treatment (median 29 days) and 33.9% underwent surgical intervention. Preoperative serum albumin <35 g/L (p = 0.04), PCI>17 (p = 0.025) and operation >8.6 h s (p = 0.001) were independent risk factors on multivariate analysis. Overall and 5-year survival was significantly reduced (p < 0.0001,p = 0.016). CRS/HIPEC remains an effective treatment modality for PC in selected patients with a comparable ECF incidence to reported elective gastrointestinal surgery rates. This study identifies multiple risk factors that should be considered in patients undergoing CRS/HIPEC. Copyright © 2016 Elsevier Ltd. All rights reserved.

Predictors of response to enteral nutrition in abdominal enterocutaneous fistula patients with Crohn's disease.

PubMed

Yan, D; Ren, J; Wang, G; Liu, S; Li, J

2014-08-01

Enterocutaneous fistula (ECF) is a serious complication of Crohn's disease (CD). Enteral nutrition (EN) is believed as one of therapeutic strategies of CD. This study is dedicated to identify predictors of response to EN in CD, which may lead to a better selection of fistula patients for this therapy. Forty-eight CD patients with ECF treated with short-peptide-based EN for 3 months were included in this study. All patients were followed up for at least 6 months. Logistic regression was performed to investigate the potential predictors of response to EN in these patients. In total, 30 out of 48 patients were confirmed with a successful closure of fistula after 3 months' EN therapy. The average closure time was 32.4±8.85 days. Inflammatory parameters (erythrocyte sedimentation rate, C-reactive protein (CRP) and platelet count) improved significantly after EN therapy in all enrolled patients. Specifically, the improvement of CRP after therapy in closed group was more important compared with that in unclosed group (P=0.035). Nutrition status (body weight, body mass index (BMI), hemoglobin, serum albumin (ALB), serum prealbumin and total protein (TP)) improved as well (P<0.05). Similarly, after treatment, the improvement of serum albumin (P=0.046) and prealbumin (P=0.006) in closed group was much more important than those in unclosed group. Logistic regression analysis discovered that a decreased CRP level and an elevated BMI level would be beneficial to the response to EN in CD patients with ECF. In CD patients with ECF, lower CRP and higher BMI are associated with higher possibility of closure after EN treatment. EN therapy can lead to a closure of ECF in a certain proportion of patients. EN therapy could also ameliorate inflammatory condition and improve nutrition status.

Role of extracytoplasmic function sigma factor PG1660 (RpoE) in the oxidative stress resistance regulatory network of Porphyromonas gingivalis

PubMed Central

Dou, Y.; Rutanhira, H.; Chen, X.; Mishra, A.; Wang, C.; Fletcher, H.M.

2018-01-01

Summary In Porphyromonas gingivalis, the protein PG1660, composed of 174 amino acids, is annotated as an extracytoplasmic function (ECF) sigma factor (RpoE homologue-σ24). Because PG1660 can modulate several virulence factors and responds to environmental signals in P. gingivalis, its genetic properties were evaluated. PG1660 is co-transcribed with its downstream gene PG1659, and the transcription start site was identified as adenine residue 54-nucleotides upstream of the ATG translation start codon. In addition to binding its own promoter, using the purified rPG1660 and RNAP core enzyme from Escherichia coli with the PG1660 promoter DNA as template, the function of PG1660 as a sigma factor was demonstrated in an in vitro transcription assay. Transcriptome analyses of a P. gingivalis PG1660-defective isogenic mutant revealed that under oxidative stress conditions 176 genes including genes involved in secondary metabolism were downregulated more than two-fold compared with the parental strain. The rPG1660 protein also showed the ability to bind to the promoters of the highly downregulated genes in the PG1660-deficient mutant. As the ECF sigma factor PG0162 has a 29% identity with PG1660 and can modulate its expression, the cross-talk between their regulatory networks was explored. The expression profile of the PG0162PG1660-deficient mutant (P. gingivalis FLL356) revealed that the type IX secretion system genes and several virulence genes were downregulated under hydrogen peroxide stress conditions. Taken together, we have confirmed that PG1660 can function as a sigma factor, and plays an important regulatory role in the oxidative stress and virulence regulatory network of P. gingivalis. PMID:29059500

Executive functions and basic symptoms in adolescent antisocial behavior: a cross-sectional study on an Italian sample of late-onset offenders.

PubMed

Muscatello, Maria Rosaria A; Scimeca, Giuseppe; Pandolfo, Gianluca; Micò, Umberto; Romeo, Vincenzo M; Mallamace, Domenico; Mento, Carmela; Zoccali, Rocco; Bruno, Antonio

2014-04-01

Executive cognitive functions (ECFs) and other cognitive impairments, such as lower IQ and verbal deficits, have been associated with the pattern of antisocial and delinquent behavior starting in childhood (early-onset), but not with late-onset antisocial behavior. Beyond objective measures of ECF, basic symptoms are prodromal, subjectively experienced cognitive, perceptual, affective, and social disturbances, associated with a range of psychiatric disorders, mainly with psychosis. The goal of the present study was to examine ECF and basic symptoms in a sample of late-onset juvenile delinquents. Two-hundred nine male adolescents (aged 15-20 years) characterized by a pattern of late-onset delinquent behavior with no antecedents of Conduct Disorder, were consecutively recruited from the Social Services of the Department of Juvenile Justice of the city of Messina (Italy), and compared with nonantisocial controls matched for age, educational level, and socio-demographic features on measures for ECF dysfunction and basic symptoms. Significant differences between late-onset offenders (completers=147) and control group (n=150) were found on ECF and basic symptoms measures. Chi-square analysis showed that a significantly greater number of late-onset offending participants scored in the clinical range on several ECF measures. Executive cognitive impairment, even subtle and subclinical, along with subjective symptoms of cognitive dysfunction (basic symptom), may be contributing factor in the development and persistence of antisocial behaviors displayed by late-onset adolescent delinquents. The findings also suggest the need for additional research aimed to assess a broader range of cognitive abilities and specific vulnerability and risk factors for late-onset adolescent offenders. Copyright © 2014 Elsevier Inc. All rights reserved.

Changes in subcellular distribution of ependymins in goldfish brain induced by learning.

PubMed

Schmidt, R

1987-06-01

Goldfish were trained for 4 h to swim with an attached polystyrene foam float and tested for retention 3 days later. Intracerebroventricular injection of anti-ependymin antisera was shown to prevent long-term memory formation of this vestibulomotor learning task, as reported previously. In further experiments, fish were killed 4-14 h after the start of training. The brains were dissected, incubated in an isoosmolar solution for collection of proteins of the brain extracellular fluid (ECF), homogenized, and fractionated by differential centrifugation. The ECF, a supernatant fraction enriched in cytoplasmic constituents (S3), and various particulate subcellular fractions were analyzed for their ependymin contents by radioimmunoassay. No statistically significant changes that might be induced by the learning were revealed in any of the particulate fractions. Steady-state concentrations of ependymins in the cytoplasm, however, increased temporarily by 39% in fish that had mastered the training task as compared with nonlearning animals (passive and active controls). In the ECF, the specific concentration of ependymins first decreased to 88% of control levels (4-5 h after the start of training), but later on, it increased to 138% (8-14 h). Apparently, ependymins present in the ECF are used during biochemical reactions of memory consolidation. The resulting decrease in extracellular ependymin concentrations might trigger their resynthesis in the cytoplasm and lead to an increased release of these glycoproteins into the ECF.

Independent predictors of enteric fistula and abdominal sepsis after damage control laparotomy: results from the prospective AAST Open Abdomen registry.

PubMed

Bradley, Matthew J; Dubose, Joseph J; Scalea, Thomas M; Holcomb, John B; Shrestha, Binod; Okoye, Obi; Inaba, Kenji; Bee, Tiffany K; Fabian, Timothy C; Whelan, James F; Ivatury, Rao R

2013-10-01

Enterocutaneous fistula (ECF), enteroatmospheric fistula (EAF), and intra-abdominal sepsis/abscess (IAS) are major challenges for surgeons caring for patients undergoing damage control laparotomy after trauma. To determine independent predictors of ECF, EAF, or IAS in patients undergoing damage control laparotomy after trauma, using the AAST Open Abdomen Registry. The AAST Open Abdomen registry of patients with an open abdomen following damage control laparotomy was used to identify patients who developed ECF, EAF, or IAS and to compare these patients with those without these complications. Univariate analyses were performed to compare these groups of patients. Variables from univariate analyses differing at P < .20 were entered into a stepwise logistic regression model to identify independent risk factors for ECF, EAF, or IAS. Fourteen level I trauma centers. A total of 517 patients with an open abdomen following damage control laparotomy. Complication of ECF, EAF, or IAS. More patients in the ECF/EAF/IAS group than in the group without these complications underwent bowel resection (63 of 111 patients [57%] vs 133 of 406 patients [33%]; P < .001). Within the first 48 hours after surgery, the ECF/EAF/IAS group received more colloids (P < .03) and total fluids (P < .03) than did the group without these complications. The ECF/EAF/IAS group underwent almost twice as many abdominal reexplorations as did the group without these complications (mean [SD] number, 4.1 [4.1] vs 2.2 [3.4]; P < .001). After multivariate analysis, the independent predictors of ECF/EAF/IAS were a large bowel resection (adjusted odds ratio [AOR], 3.56 [95% CI, 1.88-6.76]; P < .001), a total fluid intake at 48 hours of between 5 and 10 L (AOR, 2.11 [95% CI, 1.15-3.88]; P = .02) or more than 10 L (AOR, 1.93 [95% CI, 1.04-3.57]; P = .04), and an increasing number of reexplorations (AOR, 1.14 [95% CI, 1.06-1.21]; P < .001). Large bowel resection, large-volume fluid resuscitation, and an increasing number of abdominal reexplorations were statistically significant predictors of ECF, EAF, or IAS in patients with an open abdomen after damage control laparotomy.

Application of system dynamics and participatory spatial group model building in animal health: A case study of East Coast Fever interventions in Lundazi and Monze districts of Zambia

PubMed Central

Skjerve, Eystein; Rich, Magda; Rich, Karl M.

2017-01-01

East Coast Fever (ECF) is the most economically important production disease among traditional beef cattle farmers in Zambia. Despite the disease control efforts by the government, donors, and farmers, ECF cases are increasing. Why does ECF oscillate over time? Can alternative approaches such as systems thinking contribute solutions to the complex ECF problem, avoid unintended consequences, and achieve sustainable results? To answer these research questions and inform the design and implementation of ECF interventions, we qualitatively investigated the influence of dynamic socio-economic, cultural, and ecological factors. We used system dynamics modelling to specify these dynamics qualitatively, and an innovative participatory framework called spatial group model building (SGMB). SGMB uses participatory geographical information system (GIS) concepts and techniques to capture the role of spatial phenomenon in the context of complex systems, allowing stakeholders to identify spatial phenomenon directly on physical maps and integrate such information in model development. Our SGMB process convened focus groups of beef value chain stakeholders in two distinct production systems. The focus groups helped to jointly construct a series of interrelated system dynamics models that described ECF in a broader systems context. Thus, a complementary objective of this study was to demonstrate the applicability of system dynamics modelling and SGMB in animal health. The SGMB process revealed policy leverage points in the beef cattle value chain that could be targeted to improve ECF control. For example, policies that develop sustainable and stable cattle markets and improve household income availability may have positive feedback effects on investment in animal health. The results obtained from a SGMB process also demonstrated that a “one-size-fits-all” approach may not be equally effective in policing ECF in different agro-ecological zones due to the complex interactions of socio-ecological context with important, and often ignored, spatial patterns. PMID:29244862

Application of system dynamics and participatory spatial group model building in animal health: A case study of East Coast Fever interventions in Lundazi and Monze districts of Zambia.

PubMed

Mumba, Chisoni; Skjerve, Eystein; Rich, Magda; Rich, Karl M

2017-01-01

East Coast Fever (ECF) is the most economically important production disease among traditional beef cattle farmers in Zambia. Despite the disease control efforts by the government, donors, and farmers, ECF cases are increasing. Why does ECF oscillate over time? Can alternative approaches such as systems thinking contribute solutions to the complex ECF problem, avoid unintended consequences, and achieve sustainable results? To answer these research questions and inform the design and implementation of ECF interventions, we qualitatively investigated the influence of dynamic socio-economic, cultural, and ecological factors. We used system dynamics modelling to specify these dynamics qualitatively, and an innovative participatory framework called spatial group model building (SGMB). SGMB uses participatory geographical information system (GIS) concepts and techniques to capture the role of spatial phenomenon in the context of complex systems, allowing stakeholders to identify spatial phenomenon directly on physical maps and integrate such information in model development. Our SGMB process convened focus groups of beef value chain stakeholders in two distinct production systems. The focus groups helped to jointly construct a series of interrelated system dynamics models that described ECF in a broader systems context. Thus, a complementary objective of this study was to demonstrate the applicability of system dynamics modelling and SGMB in animal health. The SGMB process revealed policy leverage points in the beef cattle value chain that could be targeted to improve ECF control. For example, policies that develop sustainable and stable cattle markets and improve household income availability may have positive feedback effects on investment in animal health. The results obtained from a SGMB process also demonstrated that a "one-size-fits-all" approach may not be equally effective in policing ECF in different agro-ecological zones due to the complex interactions of socio-ecological context with important, and often ignored, spatial patterns.

Characterization and in vivo regulon determination of an ECF sigma factor and its cognate anti-sigma factor in Nostoc punctiforme.

PubMed

Bell, Nicole; Lee, Jamie J; Summers, Michael L

2017-04-01

Based on primary sequence comparisons and genomic context, Npun_F4153 (SigG)/Npun_F4154 (SapG) of the cyanobacterium Nostoc punctiforme were hypothesized to encode an ECF sigma factor/anti-sigma factor pair. Transcription of sigG increased in heterocysts and akinetes, and after EDTA treatment. Interaction between SigG and the predicted cytoplasmic domain of SapG was observed in vitro. A SigG-GFP translational fusion protein localized to the periphery of vegetative cells in vivo, but lost this association following heat stress. A sigG mutant was unable to survive envelope damage caused by heat or EDTA, but was able to form functional heterocysts. Akinetes in the mutant strain appeared normal, but these cultures were less resistant to lysozyme and cold treatments than those of the wild-type strain. The SigG in vivo regulon was determined before and during akinete differentiation using DNA microarray analysis, and found to include multiple genes with putative association to the cell envelope. Mapped promoters common to both arrays enabled identification of a SigG promoter-binding motif that was supported in vivo by reporter studies, and in vitro by run-off transcription experiments. These findings support SigG/SapG as a sigma/anti-sigma pair involved in repair of envelope damage resulting from exogenous sources or cellular differentiation. © 2017 John Wiley & Sons Ltd.

Characterization and in vivo regulon determination of an ECF sigma factor and its cognate anti-sigma factor in Nostoc punctiforme

PubMed Central

Bell, Nicole; Lee, Jamie J.; Summers, Michael L.

2017-01-01

Summary Based on primary sequence comparisons and genomic context, Npun_F4153 (SigG)/Npun_F4154 (SapG) of the cyanobacterium Nostoc punctiforme were hypothesized to encode an ECF sigma factor/anti-sigma factor pair. Transcription of sigG increased in heterocysts and akinetes, and after EDTA treatment. Interaction between SigG and the predicted cytoplasmic domain of SapG was observed in vitro. A SigG-GFP translational fusion protein localized to the periphery of vegetative cells in vivo, but lost this association following heat stress. A sigG mutant was unable to survive envelope damage caused by heat or EDTA, but was able to form functional heterocysts. Akinetes in the mutant strain appeared normal, but these cultures were less resistant to lysozyme and cold treatments than those of the wild-type strain. The SigG in vivo regulon was determined before and during akinete differentiation using DNA microarray analysis, and found to include multiple genes with putative association to the cell envelope. Mapped promoters common to both arrays enabled identification of a SigG promoter-binding motif that was supported in vivo by reporter studies, and in vitro by run-off transcription experiments. These findings support SigG/SapG as a sigma/anti-sigma pair involved in repair of envelope damage resulting from exogenous sources or cellular differentiation. PMID:28105698

Protective role of ellagitannins from Eucalyptus citriodora against ethanol-induced gastric ulcer in rats: impact on oxidative stress, inflammation and calcitonin-gene related peptide.

PubMed

Al-Sayed, Eman; El-Naga, Reem N

2015-01-15

The gastroprotective activity of an ellagitannin-rich fraction obtained from Eucalyptus citriodora (ECF) was investigated against ethanol-induced gastric ulceration in rats. The rats were pretreated with ECF (25, 50 and 100mg/kg) 1h before the administration of absolute ethanol to induce acute gastric ulceration. The gastric lesions were significantly reduced by all doses of ECF. Notably, pre-treatment with ECF (100mg/kg) conferred 99.6% gastroprotection, which is significantly higher than that produced by omeprazole. Moreover, ECF administration markedly increased the mucin content in a dose-dependent manner. The potent gastroprotective effect of ECF could be partly mediated by attenuating ethanol-induced oxidative stress. ECF-pre-treatment markedly increased the depleted GSH and SOD levels in a dose-dependent manner. Moreover, ECF significantly decreased the elevated MDA tissue levels induced by ethanol administration. The results demonstrated that ECF administration exerted a powerful anti-inflammatory activity as evidenced by the reduction in the pro-inflammatory markers; IL-1β, TNF-α, 5-LO and COX-2. Additionally, the caspase-3 tissue levels were significantly reduced in the groups pre-treated with ECF. These results suggest that ECF could exert a beneficial gastroprotective effect through their antioxidant, anti-inflammatory and anti-apoptotic properties. Furthermore, ECF pre-treatment significantly attenuated the ethanol-induced decrease in CGRP expression, which has a protective role against gastric ulceration. Histopathological examination revealed intact mucosal layer, absence of hemorrhage and necrosis in groups treated with ECF. Ellagitannins were identified as the major active constituents responsible for the marked antioxidant and gastroprotective properties of ECF. The HPLC-PDA-ESI/MS/MS technique was employed to identify the ellagitannins of E. citriodora. Copyright © 2014 Elsevier GmbH. All rights reserved.

The ECF sigma factor, PSPTO_1043, in Pseudomonas syringae pv. tomato DC3000 is induced by oxidative stress and regulates genes involved in oxidative stress response

PubMed Central

Butcher, Bronwyn G.; Bao, Zhongmeng; Wilson, Janet; Swingle, Bryan; Filiatrault, Melanie; Schneider, David; Cartinhour, Samuel

2017-01-01

The bacterial plant pathogen Pseudomonas syringae adapts to changes in the environment by modifying its gene expression profile. In many cases, the response is mediated by the activation of extracytoplasmic function (ECF) sigma factors that direct RNA polymerase to transcribe specific sets of genes. In this study we focus on PSPTO_1043, one of ten ECF sigma factors in P. syringae pv. tomato DC3000 (DC3000). PSPTO_1043, together with PSPTO_1042, encode an RpoERsp/ChrR-like sigma/anti-sigma factor pair. Although this gene pair is unique to the P. syringae group among the pseudomonads, homologous genes can be found in photosynthetic genera such as Rhodospirillum, Thalassospira, Phaeospirillum and Parvibaculum. Using ChIP-Seq, we detected 137 putative PSPTO_1043 binding sites and identified a likely promoter motif. We characterized 13 promoter candidates, six of which regulate genes that appear to be found only in P. syringae. PSPTO_1043 responds to the presence of singlet oxygen (1O2) and tert-butyl hydroperoxide (tBOOH) and several of the genes regulated by PSPTO_1043 appear to be involved in response to oxidative stress. PMID:28700608

The ECF sigma factor, PSPTO_1043, in Pseudomonas syringae pv. tomato DC3000 is induced by oxidative stress and regulates genes involved in oxidative stress response.

PubMed

Butcher, Bronwyn G; Bao, Zhongmeng; Wilson, Janet; Stodghill, Paul; Swingle, Bryan; Filiatrault, Melanie; Schneider, David; Cartinhour, Samuel

2017-01-01

The bacterial plant pathogen Pseudomonas syringae adapts to changes in the environment by modifying its gene expression profile. In many cases, the response is mediated by the activation of extracytoplasmic function (ECF) sigma factors that direct RNA polymerase to transcribe specific sets of genes. In this study we focus on PSPTO_1043, one of ten ECF sigma factors in P. syringae pv. tomato DC3000 (DC3000). PSPTO_1043, together with PSPTO_1042, encode an RpoERsp/ChrR-like sigma/anti-sigma factor pair. Although this gene pair is unique to the P. syringae group among the pseudomonads, homologous genes can be found in photosynthetic genera such as Rhodospirillum, Thalassospira, Phaeospirillum and Parvibaculum. Using ChIP-Seq, we detected 137 putative PSPTO_1043 binding sites and identified a likely promoter motif. We characterized 13 promoter candidates, six of which regulate genes that appear to be found only in P. syringae. PSPTO_1043 responds to the presence of singlet oxygen (1O2) and tert-butyl hydroperoxide (tBOOH) and several of the genes regulated by PSPTO_1043 appear to be involved in response to oxidative stress.

Improvement of Output Power of ECF Micromotor

NASA Astrophysics Data System (ADS)

Yokota, Shinichi; Kawamura, Kiyomi; Takemura, Kenjiro; Edamura, Kazuya

Electro-conjugate fluid (ECF) is a kind of dielectric fluids, which produces jet-flow (ECF jet) when subjected to a high DC voltage. By using the ECF jet, a new type of micromotor with simple structure and lightweight can be realized. Up to now, we developed a disk-plate type ECF micromotor with inner diameter of 9 mm. In this study, we develope novel ECF micromotors with inner diameter of 5 mm in order to improve the output power density. First, we designed and produced the ECF micromotors with 4-layered and 8-layered disk plate rotors. Then, the performances of the motors are measured. The experimental results confirm the motor developed has a higher performance than the previous ones.

Extracardiac findings at cardiac MR imaging: a single-centre retrospective study over 14 years.

PubMed

Sokolowski, Felix C; Karius, Philipp; Rodríguez, Alejandra; Lembcke, Alexander; Wagner, Moritz; Hamm, Bernd; Dewey, Marc

2018-04-30

To determine the prevalence and significance of extracardiac findings (ECF) in a large set of cardiac magnetic resonance (MR) imaging examinations. The institutional review board (IRB) of the Charité approved this retrospective, single-centre study. A total of 4376 cardiac MR imaging reports of 3553 patients (age 37.4 ± 20 years, 60.8 % male) examined from 2000 to 2014 were included. Findings with a recommendation for follow-up were considered "major ECF". To analyse the association of indication, age and gender with ECF, Poisson regression and computed incidence rate ratios (IRR) were evaluated. The overall prevalence of ECF was 34% (95% confidence interval [CI] 32.5-35.6%). Major ECF were present in 3.4% (95% CI 2.9-4.1%) while findings that changed patient management were found in 0.9% (95% CI 0.7-1.3%). In the cases of congenital heart disease, ECF prevalence was higher compared to myocarditis (IRR, 6.0; 95% CI 5.1-7.1%; p < 0.001), while the prevalence of major ECF was lower (IRR, 0.2; 95% CI 0.02-0.51%; p < 0.05). Older patient age was associated with more nonvascular ECF (p < 0.001). Female patients had the same probability of having an ECF as male patients (IRR, 1.04; 95% CI 0.95-1.1%; p = 0.43). ECF in cardiac MR imaging are present in about every third patient while relevant ECF that change patient management can be found in about one out of 100 patients. Our data suggest that it is important to involve well-trained radiologists in reading cardiac MR images, which often reveal ECF if congenital heart disease is the clinical indication. • Extracardiac findings are present in about every third patient. • Relevant ECF changing patient management are found in one out of 100 findings. • Chance of ECF is high in patients with CHD and vascular indications.

An Aromatic Cap Seals the Substrate Binding Site in an ECF-Type S Subunit for Riboflavin

DOE Office of Scientific and Technical Information (OSTI.GOV)

Karpowich, Nathan K.; Song, Jinmei; Wang, Da-Neng

2016-06-13

ECF transporters are a family of active membrane transporters for essential micronutrients, such as vitamins and trace metals. Found exclusively in archaea and bacteria, these transporters are composed of four subunits: an integral membrane substrate-binding subunit (EcfS), a transmembrane coupling subunit (EcfT), and two ATP-binding cassette ATPases (EcfA and EcfA'). We have characterized the structural basis of substrate binding by the EcfS subunit for riboflavin from Thermotoga maritima, TmRibU. TmRibU binds riboflavin with high affinity, and the protein–substrate complex is exceptionally stable in solution. The crystal structure of riboflavin-bound TmRibU reveals an electronegative binding pocket at the extracellular surface inmore » which the substrate is completely buried. Analysis of the intermolecular contacts indicates that nearly every available substrate hydrogen bond is satisfied. A conserved aromatic residue at the extracellular end of TM5, Tyr130, caps the binding site to generate a substrate-bound, occluded state, and non-conservative mutation of Tyr130 reduces the stability of this conformation. Using a novel fluorescence binding assay, we find that an aromatic residue at this position is essential for high-affinity substrate binding. Comparison with other S subunit structures suggests that TM5 and Loop5-6 contain a dynamic, conserved motif that plays a key role in gating substrate entry and release by S subunits of ECF transporters.« less

East Coast fever immunisation field trial in crossbred dairy cattle in Hanang and Handeni districts in northern Tanzania.

PubMed

Lynen, Godelieve; Yrjö-Koskinen, Alma E; Bakuname, Christine; Di Giulio, Giuseppe; Mlinga, Nevil; Khama, Isaac; Hanks, James; Taylor, Nick M; James, Andrew D; McKeever, Declan; Peters, Andy R; Rushton, Jonathan

2012-03-01

East Coast fever (ECF) causes considerable mortality and production losses in the Tanzania smallholder dairy sector and limits the introduction of improved dairy breeds in areas where the disease is present. The infection and treatment method (ITM) was adopted by smallholder dairy farms for ECF immunisation in Hanang and Handeni districts of Tanzania. This study recorded incidence rates for ECF and other tick-borne diseases (TBDs) for ECF-immunised and non-immunised cattle between 1997 and 2000. Approximately 80% of smallholder households from both sites (n = 167) participated in this longitudinal study, with immunisations carried out at the request of the livestock owners. Efficacy of ITM for preventing ECF cases in these crossbred dairy cattle was estimated at 97.6%, while that for preventing ECF deaths was 97.9%. One percent of the cattle developed clinical ECF as a result of immunisation. Since ECF immunisation permits a reduction in acaricide use, an increase in other TBDs is a potential concern. Sixty-three percent of farmers continued to use the same acaricide after immunisation, with 80% of these reducing the frequency of applications. Overall, 78% of farmers increased the acaricide application interval after immunisation beyond that recommended by the manufacturer, resulting in annual savings in the region of USD 4.77 per animal. No statistical difference was observed between the immunised and non-immunised animals in the incidence of non-ECF TBDs. However, immunised animals that succumbed to these diseases showed fewer case fatalities. ITM would therefore appear to be a suitable method for ECF control in Tanzania's smallholder dairy sector.

The plasticity of extracellular fluid homeostasis in insects.

PubMed

Beyenbach, Klaus W

2016-09-01

In chemistry, the ratio of all dissolved solutes to the solution's volume yields the osmotic concentration. The present Review uses this chemical perspective to examine how insects deal with challenges to extracellular fluid (ECF) volume, solute content and osmotic concentration (pressure). Solute/volume plots of the ECF (hemolymph) reveal that insects tolerate large changes in all three of these ECF variables. Challenges beyond those tolerances may be 'corrected' or 'compensated'. While a correction simply reverses the challenge, compensation accommodates the challenge with changes in the other two variables. Most insects osmoregulate by keeping ECF volume and osmotic concentration within a wide range of tolerance. Other insects osmoconform, allowing the ECF osmotic concentration to match the ambient osmotic concentration. Aphids are unique in handling solute and volume loads largely outside the ECF, in the lumen of the gut. This strategy may be related to the apparent absence of Malpighian tubules in aphids. Other insects can suspend ECF homeostasis altogether in order to survive extreme temperatures. Thus, ECF homeostasis in insects is highly dynamic and plastic, which may partly explain why insects remain the most successful class of animals in terms of both species number and biomass. © 2016. Published by The Company of Biologists Ltd.

75 FR 48615 - Airworthiness Directives; Eurocopter France (ECF) Model AS350B3 and EC130 B4 Helicopters

Federal Register 2010, 2011, 2012, 2013, 2014

2010-08-11

.... Related Service Information ECF has issued Alert Service Bulletin (ASB) No. 05.00.61 for the Model AS350B3... Airworthiness Directives; Eurocopter France (ECF) Model AS350B3 and EC130 B4 Helicopters AGENCY: Federal... adopt a new airworthiness directive (AD) for the specified ECF model helicopters. This proposed AD...

[Extracellular fluid, plasma and interstitial volume in cirrhotic patients without clinical edema or ascites].

PubMed

Noguera Viñas, E C; Hames, W; Mothe, G; Barrionuevo, M P

1989-01-01

Extracellular fluid volume (E.C.F.) and plasma volume (P.V.), were measured with sodium sulfate labeled with 35I and 131I human serum albumin, respectively, by the dilution technique in control subjects and in cirrhotic patients without clinical ascites or edema, renal or hepatic failure, gastrointestinal bleeding or diuretics. Results are expressed as mean +/- DS in both ml/m2 and ml/kg. In normal subjects E.C.F. (n = 8) was 7,533 +/- 817 ml/m2 (201.3 +/- 182 ml/kg), P.V. (n = 11) 1,767 +/- 337 ml/m2 (47.2 +/- 9.3 ml/kg), and interstitial fluid (I.S.F.) (n = 7) 5,758 +/- 851 ml/m2 (Table 2). In cirrhotic patients E.C.F. (n = 11) was 10,318 +/- 2,980 ml/m2 (261.7 +/- 76.8 ml/kg), P.V. (n = 12) 2,649 +/- 558 ml/m2 (67.7 +/- 15.6 ml/kg) and I.S.F. (n = 11) 7,866 +/- 2,987 ml/m2 (Table 3). Cirrhotic patients compared with normal subjects have hypervolemia due to a significant E.C.F. and P.V. expansion (p less than 0.02 and less than 0.001 respectively) (Fig. 1). Reasons for E.C.F. and P.V. abnormalities in cirrhotic patients may reflect urinary sodium retention related to portal hipertension which stimulates aldosterone release or enhanced renal tubular sensitivity to the hormone. However, it is also possible that these patients, in the presence of hypoalbuminemia (Table 1), have no clinical edema or ascites due to increased glomerular filtration, suppressed release of vasopressin, increased natriuretic factor, and urinary prostaglandin excretion, in response to the intravascular expansion, all of which increased solute and water delivery to the distal nephron and improved renal water excretion. We conclude that in our clinical experience cirrhotic patients without ascites or edema have hypervolemia because of a disturbance in E.C.F.

A ten-year review of enterocutaneous fistulas after laparotomy for trauma.

PubMed

Fischer, Peter E; Fabian, Timothy C; Magnotti, Louis J; Schroeppel, Thomas J; Bee, Tiffany K; Maish, George O; Savage, Stephanie A; Laing, Ashley E; Barker, Andrew B; Croce, Martin A

2009-11-01

In the era of open abdomen management, the complication of enterocutaneous fistula (ECF) seems to be increasing in frequency. In nontrauma patients, reported mortality rates are 7% to 20%, and spontaneous closure rates are approximately 25%. This study is the largest series of ECFs reported exclusively caused by trauma and examines the characteristics unique to this population. Trauma patients with an ECF at a single regional trauma center over a 10-year period were reviewed. Parameters studied included fistula output, site, nutritional status, operative history, and fistula resolution (spontaneous vs. operative). Approximately 2,224 patients received a trauma laparotomy and survived longer than 4 days. Of these, 43 patients (1.9%) had ECF. The rate of ECF in men was 2.22% and 0.74% in women. Patients with open abdomen had a higher ECF incidence (8% vs. 0.5%) and lower rate of spontaneous closure (37% vs. 45%). Spontaneous closure occurred in 31% with high-output fistulas, 13% with medium output, and 55% with low output. The mortality rate of ECF was 14% after an average stay of 59 days in the intensive care unit. With damage-control laparotomies, the traumatic ECF rate is increasing and is a different entity than nontraumatic ECF. Although the two populations have similar mortality rates, the trauma cohort demonstrates higher spontaneous closure rates and a curiously higher rate of development in men. Fistula output was not predictive of spontaneous closure.

The current epidemiological status of bovine theileriosis in eastern Zambia.

PubMed

Billiouw, M; Mataa, L; Marcotty, T; Chaka, G; Brandt, J; Berkvens, D

1999-09-01

Results of a longitudinal study conducted in the eastern province of Zambia from 1994 to 1997 indicate that it is doubtful whether a state of endemic stability of East Coast fever (ECF) can be reached in the near future. Even in endemic areas, the mortality of Theileria parva infections is still estimated above 50%. The main factors limiting progress towards endemic stability are high innate susceptibility of the Zebu cattle, the virulence of the parasite and the climate. The unimodal rainfall pattern results in restricted activity of Rhipicephalus appendiculatus instars and year-to-year variation in rainfall causes fluctuations in tick phenology and T. parva transmission. Adult tick activity invariably peaks during the rains and is associated with the highest ECF incidence. Nymphal transmission of T. parva to cattle appears to be less important. Second periods of activity of both adult and nymphal instars are pronounced only when the climate is suitable. These second waves of tick activity ensure a more continuous and efficient transmission of T. parva and also play a key role in the dynamics of prolonged outbreaks in epidemic areas. ECF control methods may have an important influence on ECF epidemiology. Immunizations as well as chemotherapy of clinical cases create a reservoir of virulent parasites in susceptible cattle, resulting in artificial endemic stability.

Immunity to East Coast fever in cattle induced by a polypeptide fragment of the major surface coat protein of Theileria parva sporozoites.

PubMed

Bishop, Richard; Nene, Vishvanath; Staeyert, Jan; Rowlands, John; Nyanjui, John; Osaso, Julius; Morzaria, Subhash; Musoke, Antony

2003-03-07

Full-length recombinant versions of p67, the 709 amino acid major surface protein of Theileria parva sporozoites, induce immunity to East Coast fever (ECF) in cattle. We show that a soluble Escherichia coli recombinant version of p67 (p67(635)), in which a prokaryotic signal peptide replaces the eukaryotic one, confers protection comparable to that induced by the full-length molecule, but is unstable. Peptides encoding 80 (p67C) and 205 (p67N) amino acid fragments of p67, containing epitopes recognised by sporozoite neutralising monoclonal antibodies, exhibit improved stability in E. coli. Antibodies raised against the central region of p67 (p67M) neutralise sporozoite infectivity in vitro. The p67C peptide induced immunity against ECF in cattle, at a level equivalent to p67(635), suggesting that a synthetic peptide vaccine might be achievable. Copyright 2002 Elsevier Science Ltd.

Identification of a target population for immunisation against East Coast fever in coastal Kenya.

PubMed

Maloo, S H; Ngumi, P; Mbogo, S; Williamson, S; Thorpe, W; Rowlands, G J; Perry, B D

2001-11-02

Two experiments were carried out to identify the target population of cattle for immunisation against East Coast fever (ECF) using the infection-and-treatment method. Firstly, a sentinel-calf study was used to determine the age window for ECF immunisation by determining ages at clinical detection of infection with Theileria parva. Six groups of five naive cross-bred (Bos taurus/Bos indicus) male calves, introduced at intervals of 2 months at a mean age of 26 days, were exposed to natural tick challenge on a high ECF-risk, small-holder farm in the coastal lowland, coconut-cassava agro-ecological zone of coastal Kenya. Secondly, a challenge study evaluated the relationship between the presence of T. parva antibodies and immunity. Ten indigenous adult Zebu cattle and nine Zebu young stock purchased from farmers in the same zone, and eight cross-bred calves (survivors of the sentinel-calf study) were challenged with 10 times the immunising dose of T. parva Marikebuni stock. Twenty-four of these 27 cattle had high antibody titres before challenge. Two cross-bred calves, obtained from an ECF-free area and seronegative to T. parva schizont antigen, also were challenged and used as susceptible controls. Twenty-five (83%) of the 30 sentinel calves contracted ECF over an age range of 36-116 days (mean 72 days). The remaining five calves died of other causes within 2 months of arrival on the farm. Fourteen of the 25 calves survived the infection and developed antibodies to T. parva. Despite tick control, seven of these 14 calves had a second episode of ECF and two died. In total, 13 of the 25 calves that contracted ECF died. Only one of 19 indigenous Zebu animals developed clinical ECF when challenged with T. parva Marikebuni (mild clinical signs with spontaneous recovery). Of the eight cross-bred survivors from the first experiment, only one succumbed to ECF when challenged and it died. Both susceptible cross-bred calves developed severe clinical signs of ECF and one died. The experimental studies show that in the high ECF-risk areas of the coconut-cassava zone of coastal Kenya, immunisation against ECF in cross-bred (B. taurus/B. indicus) cattle should be targeted at an early age (preferably within 1-2 months of birth).

UV-LIGA technique for ECF micropumps using back UV exposure and self-alignment

NASA Astrophysics Data System (ADS)

Han, D.; Xia, Y.; Yokota, S.; Kim, J. W.

2017-12-01

This paper proposes and develops a novel UV-LIGA technique using back UV exposure and self-alignment to realize high aspect ratio micromachining (HARM) in high power density electro-conjugate fluid (ECF) micropumps. ECF is a functional fluid designed to be able to generate strong and active jet flow (ECF jetting) between anode and cathode in ECF when high DC voltage is applied. We have developed high power density ECF micropumps consisting of triangular prism and slit electrode pairs (TPSEs) fabricated by HARM. The traditional UV-LIGA technique for HARM is mainly divided into two approaches: (a) single thick layer and (b) multiple thin layers. Both methods have limitations—deformed molds in the former and misalignment between layers in the latter. Using the finite element method software COMSOL Multiphysics, we demonstrate that the deformed micro-molds critically impair the performance of ECF micropumps. In addition, we experimentally prove that the misalignment would easily trigger electric discharge in the ECF micropumps. To overcome these limitations, we conceive a new concept utilizing the seed electrode layer for electroforming as the UV shield and pattern photoresist (KMPR) by back UV exposure. The seed electrode layer should be composed of a non-transparent conductor (Au/Ti) for patterning and a transparent conductor (ITO) for wiring. Instead of ITO, we propose the concept of transparency-like electrodes comprised of thin metal line patterns. To verify this concept, KMPR layers with thicknesses of 70, 220, and 500 µm are experimentally investigated. In the case of 500 µm KMPR thickness, the concept of transparency-like electrode was partially proved. As a result, TPSEs with a height of 440 µm were successfully fabricated. Characteristic experiments demonstrated that ECF micropumps (367 mW cm-3) fabricated by back UV achieved almost the same output power density as ECF micropumps (391 mW cm-3) fabricated by front UV. This paper proves that the proposed UV-LIGA technique, using back UV exposure and self-alignment, can effectively fabricate TPSEs for ECF micropumps with high output power density.

Treatment Strategies in 135 Consecutive Patients with Enterocutaneous Fistulas

PubMed Central

Visschers, Ruben G. J.; Damink, Steven W. M. Olde; Winkens, Bjorn; Soeters, Peter B.

2008-01-01

Background Enterocutaneous fistulas (ECF) pose a major challenge to every gastrointestinal (GI) surgeon. Based on earlier studies, a standardized treatment guideline was implemented. The focus of the present study was to assess that guideline and determine prognostic factors for outcome of patients with ECF, and to define a more detailed therapeutic approach including the convalescence time before restorative surgery. Methods All patients with ECF treated between 1990 and 2005 were included. Management consisted of controlling Sepsis, Optimization of nutritional state, Wound care, assessment of fistula Anatomy, Timing of surgery, and Surgical strategy (the SOWATS guideline). Prognostic factors were assessed by way of multiple logistic regression analysis. Results A total of 135 patients were treated at our unit. Overall closure was achieved in 118 patients (87.4%). Restorative operations for fistula closure were performed after a median of 53 days (range: 4–270 days). Restorative operations were successful in 97/107 patients (90.7%). Thirteen patients (9.6%) died. An abdominal wall defect was the most predominant negative prognostic factor for spontaneous closure (odds ratio [OR] = 0.195, confidence interval [CI] 0.052–0.726, p = 0.015). A strong relation was found between preoperative albumin level and surgical closure (p < 0.001) and mortality (p < 0.001). Conclusions Application of the SOWATS guideline allowed a favorable outcome after a short convalescence period. Abdominal wall defects and preoperative hypoalbuminemia are important prognostic variables. PMID:18175171

Treatment strategies in 135 consecutive patients with enterocutaneous fistulas.

PubMed

Visschers, Ruben G J; Olde Damink, Steven W M; Winkens, Bjorn; Soeters, Peter B; van Gemert, Wim G

2008-03-01

Enterocutaneous fistulas (ECF) pose a major challenge to every gastrointestinal (GI) surgeon. Based on earlier studies, a standardized treatment guideline was implemented. The focus of the present study was to assess that guideline and determine prognostic factors for outcome of patients with ECF, and to define a more detailed therapeutic approach including the convalescence time before restorative surgery. All patients with ECF treated between 1990 and 2005 were included. Management consisted of controlling Sepsis, Optimization of nutritional state, Wound care, assessment of fistula Anatomy, Timing of surgery, and Surgical strategy (the SOWATS guideline). Prognostic factors were assessed by way of multiple logistic regression analysis. A total of 135 patients were treated at our unit. Overall closure was achieved in 118 patients (87.4%). Restorative operations for fistula closure were performed after a median of 53 days (range: 4-270 days). Restorative operations were successful in 97/107 patients (90.7%). Thirteen patients (9.6%) died. An abdominal wall defect was the most predominant negative prognostic factor for spontaneous closure (odds ratio [OR]=0.195, confidence interval [CI] 0.052-0.726, p=0.015). A strong relation was found between preoperative albumin level and surgical closure (p<0.001) and mortality (p<0.001). Application of the SOWATS guideline allowed a favorable outcome after a short convalescence period. Abdominal wall defects and preoperative hypoalbuminemia are important prognostic variables.

Bacillus subtilis extracytoplasmic function (ECF) sigma factors and defense of the cell envelope

PubMed Central

Helmann, John D.

2016-01-01

Summary Bacillus subtilis provides a model for investigation of the bacterial cell envelope, the first line of defense against environmental threats. Extracytoplasmic function (ECF) sigma factors activate genes that confer resistance to agents that threaten the integrity of the envelope. Although their individual regulons overlap, σW is most closely associated with membrane-active agents, σX with cationic antimicrobial peptide resistance, and σV with resistance to lysozyme. Here, I highlight the role of the σM regulon, which is strongly induced by conditions that impair peptidoglycan synthesis and includes the core pathways of envelope synthesis and cell division, as well as stress-inducible alternative enzymes. Studies of these cell envelope stress responses provide insights into how bacteria acclimate to the presence of antibiotics. PMID:26901131

Therapeutic potential of Pirfenidone for treating equine corneal scarring

PubMed Central

Fink, Michael K.; Giuliano, Elizabeth A.; Tandon, Ashish; Mohan, Rajiv R.

2014-01-01

Objective To evaluate the safety and efficacy of Pirfenidone (PFD) in the treatment of equine corneal fibrosis using an in vitro model. Methods Healthy donor equine corneas were collected and used to generate primary equine corneal fibroblasts (ECFs) by growing cultures in minimal essential medium supplemented with 10% fetal bovine serum. Equine corneal myofibroblasts (ECMs), used as a model of equine corneal fibrosis, were produced by growing ECF cultures in serum-free medium containing transforming growth factor β1 (1ng/ml). Trypan blue viability assays and changes in ECF morphology were utilized to determine the optimal PFD dose for this in vitro model. Trypan blue viability, phase contrast microscopy, and TUNEL assays were used to evaluate the cytotoxicity of PFD. Scratch and MTT assays were used to evaluate the effect of PFD on cellular migration and proliferation. Real-time PCR, immunoblot analysis, and immunocytochemistry were employed to determine the efficacy of PFD to inhibit ECM formation in vitro. Results Topical PFD application at 200 μg/ml successfully decreased αSMA expression when compared to the TGFβ1 only treatment group (P < 0.01). PFD application ≤ 200 μg/ml did not affect ECF phenotype or cellular viability and did not result in significant cytotoxicity. Conclusions Pirfenidone safely and effectively inhibits TGFβ1-induced equine corneal fibrosis in vitro. In vivo studies are warranted. PMID:25041235

Transcriptional regulation of the operon encoding stress-responsive ECF sigma factor SigH and its anti-sigma factor RshA, and control of its regulatory network in Corynebacterium glutamicum

PubMed Central

2012-01-01

Background The expression of genes in Corynebacterium glutamicum, a Gram-positive non-pathogenic bacterium used mainly for the industrial production of amino acids, is regulated by seven different sigma factors of RNA polymerase, including the stress-responsive ECF-sigma factor SigH. The sigH gene is located in a gene cluster together with the rshA gene, putatively encoding an anti-sigma factor. The aim of this study was to analyze the transcriptional regulation of the sigH and rshA gene cluster and the effects of RshA on the SigH regulon, in order to refine the model describing the role of SigH and RshA during stress response. Results Transcription analyses revealed that the sigH gene and rshA gene are cotranscribed from four sigH housekeeping promoters in C. glutamicum. In addition, a SigH-controlled rshA promoter was found to only drive the transcription of the rshA gene. To test the role of the putative anti-sigma factor gene rshA under normal growth conditions, a C. glutamicum rshA deletion strain was constructed and used for genome-wide transcription profiling with DNA microarrays. In total, 83 genes organized in 61 putative transcriptional units, including those previously detected using sigH mutant strains, exhibited increased transcript levels in the rshA deletion mutant compared to its parental strain. The genes encoding proteins related to disulphide stress response, heat stress proteins, components of the SOS-response to DNA damage and proteasome components were the most markedly upregulated gene groups. Altogether six SigH-dependent promoters upstream of the identified genes were determined by primer extension and a refined consensus promoter consisting of 45 original promoter sequences was constructed. Conclusions The rshA gene codes for an anti-sigma factor controlling the function of the stress-responsive sigma factor SigH in C. glutamicum. Transcription of rshA from a SigH-dependent promoter may serve to quickly shutdown the SigH-dependent stress response after the cells have overcome the stress condition. Here we propose a model of the regulation of oxidative and heat stress response including redox homeostasis by SigH, RshA and the thioredoxin system. PMID:22943411

Transcriptional regulation of the operon encoding stress-responsive ECF sigma factor SigH and its anti-sigma factor RshA, and control of its regulatory network in Corynebacterium glutamicum.

PubMed

Busche, Tobias; Silar, Radoslav; Pičmanová, Martina; Pátek, Miroslav; Kalinowski, Jörn

2012-09-03

The expression of genes in Corynebacterium glutamicum, a Gram-positive non-pathogenic bacterium used mainly for the industrial production of amino acids, is regulated by seven different sigma factors of RNA polymerase, including the stress-responsive ECF-sigma factor SigH. The sigH gene is located in a gene cluster together with the rshA gene, putatively encoding an anti-sigma factor. The aim of this study was to analyze the transcriptional regulation of the sigH and rshA gene cluster and the effects of RshA on the SigH regulon, in order to refine the model describing the role of SigH and RshA during stress response. Transcription analyses revealed that the sigH gene and rshA gene are cotranscribed from four sigH housekeeping promoters in C. glutamicum. In addition, a SigH-controlled rshA promoter was found to only drive the transcription of the rshA gene. To test the role of the putative anti-sigma factor gene rshA under normal growth conditions, a C. glutamicum rshA deletion strain was constructed and used for genome-wide transcription profiling with DNA microarrays. In total, 83 genes organized in 61 putative transcriptional units, including those previously detected using sigH mutant strains, exhibited increased transcript levels in the rshA deletion mutant compared to its parental strain. The genes encoding proteins related to disulphide stress response, heat stress proteins, components of the SOS-response to DNA damage and proteasome components were the most markedly upregulated gene groups. Altogether six SigH-dependent promoters upstream of the identified genes were determined by primer extension and a refined consensus promoter consisting of 45 original promoter sequences was constructed. The rshA gene codes for an anti-sigma factor controlling the function of the stress-responsive sigma factor SigH in C. glutamicum. Transcription of rshA from a SigH-dependent promoter may serve to quickly shutdown the SigH-dependent stress response after the cells have overcome the stress condition. Here we propose a model of the regulation of oxidative and heat stress response including redox homeostasis by SigH, RshA and the thioredoxin system.

Effect of hot-water extraction (HWE) severity on bleached pulp based biorefinery performance of eucalyptus during the HWE-Kraft-ECF bleaching process.

PubMed

Liu, Jing; Li, Meng; Luo, Xiaolin; Chen, Lihui; Huang, Liulian

2015-04-01

The effectiveness of a biorefinery based on an HWE-Kraft-ECF bleaching process and the end use of pulp was systematically evaluated. Using a P-factor of 198, nearly 30% of xylan-based sugars were recovered. The resulting pulp and paper properties were found to be comparable with the control. A maximum xylan-based sugar recovery of nearly 50% was achieved at a P-factor of 738. Although the strength of this P-factor induced handsheet was lower than that of the control by about 20%, the corresponding pulp was sufficient for dissolving pulp application. However, once the P-factor rose above 1189, hemicellulose sugars were significantly degraded into furans; pulp and paper properties were also deteriorated due to cellulose degradation, lignin deposition and condensation. Thus, considering the different end use of pulps, the performance of an HWE-based biorefinery could be balanced by its HWE severity. Copyright © 2015 Elsevier Ltd. All rights reserved.

Impairment of Executive Cognitive Control in Type 2 Diabetes, and its Effects on Health-Related Behavior and Use of Health Services

PubMed Central

Tran, Dinh; Baxter, Judith; Hamman, Richard F.; Grigsby, Jim

2013-01-01

We evaluated whether, among persons with type 2 diabetes: 1) impaired executive cognitive functioning (ECF) is more common than among people without diabetes; 2) ECF is associated with the capacity to engage in instrumental health-related behaviors; and 3) worse ECF is associated with increased health services utilization. A population-based sample of 1,063 older people was interviewed regarding medical history and health services utilization; participants were administered the Mini Mental State Exam and the Behavioral Dyscontrol Scale, a measure of ECF. Participants with diabetes performed more poorly on cognitive measures than those without diabetes. Among those with diabetes, lower ECF was associated with more outpatient care and with ever having been in a nursing home. Impaired behavioral self-regulation may affect the capacity to engage in behaviors that could improve clinical status, resulting in greater health services use. The findings suggest the possibility of a positive feedback loop, with ECF deficits adversely affecting adherence, in turn leading to greater cognitive impairment—an issue for future research. PMID:23430355

Can Structural MRI Indices of Cerebral Integrity Track Cognitive Trends in Executive Control Function During Normal Maturation and Adulthood?

PubMed Central

Kochunov, Peter; Robin, Don A.; Royall, Don R.; Coyle, Thomas; Lancaster, Jack; Kochunov, Valeria; Schlosser, Anita E.; Fox, Peter T.

2009-01-01

We explored the relationship between structural neuroimaging-based indices of cerebral integrity and executive control function (ECF) in two groups of healthy subjects: A maturing group (33 subjects; 19–29 years) and a senescing group (38 adults; 30–90 years). ECF was assessed using the Executive Interview (EXIT) battery. Cortical indices of cerebral integrity included GM thickness, intergyral span, and sulcal span, each measured for five cortical regions per hemisphere. Subcortical indices included fractional anisotropy (FA), measured using track-based-spatial-statistics (TBSS), and the volume of T2-hyperintense WM (HWM). In the maturing group, no significant relationships between neuroanatomical changes and ECF were found; however, there were hints that late-term maturation of cerebral WM influenced variability in ECF. In the senescing group, the decline in ECF corresponded to atrophic changes in cerebral WM (sulcal and intergyral span) primarily in the superior frontal and anterior cingulate regions. A large fraction of the variability in ECF (62%) can be explained by variability in the structural indices from these two regions. PMID:19067326

Bacillus subtilis extracytoplasmic function (ECF) sigma factors and defense of the cell envelope.

PubMed

Helmann, John D

2016-04-01

Bacillus subtilis provides a model for investigation of the bacterial cell envelope, the first line of defense against environmental threats. Extracytoplasmic function (ECF) sigma factors activate genes that confer resistance to agents that threaten the integrity of the envelope. Although their individual regulons overlap, σ(W) is most closely associated with membrane-active agents, σ(X) with cationic antimicrobial peptide resistance, and σ(V) with resistance to lysozyme. Here, I highlight the role of the σ(M) regulon, which is strongly induced by conditions that impair peptidoglycan synthesis and includes the core pathways of envelope synthesis and cell division, as well as stress-inducible alternative enzymes. Studies of these cell envelope stress responses provide insights into how bacteria acclimate to the presence of antibiotics. Copyright © 2016 Elsevier Ltd. All rights reserved.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Alvarez, P; Gonzalez, S; McDonald, L

Purpose: Analysis of the performance of the microSTARii reader for optically stimulated luminescence dosimeters (OSLD) used by the IROC Houston Quality Assurance Center (IROC HQAC) for external audits compare to the characteristics of the Microstar reader. Methods: The IROC HQAC uses the Microstar reader for the OSLD program for verification of output of photon, electron and proton beams. The calculation of dose from the OSLD system is based on a group of factors defined at time of the commissioning of a batch of detectors. Factors like system sensitivity (SS), depletion (KD), element correction factor (ECF), linearity (KL), energy correction (KE).more » The new microSTARii unit presents many hardware and software upgrades that were considered useful for this program. Based on these changes many factors, that were considered reader dependent, were revised in order to analyze the effect of the new reading process. The SS, KD, ECF and KL were evaluated and compared with data defined based on reading done on Microstar reader. Results: The SS is reader specific and specified at 100 cGy dose level. This value is define per reading session and monitored over time. The KD factor was found to be different because of reading procedure are different. The ECF values changed for a group of nano dots compare to values defined based on reading done on the Microstar reader. The KL was defined for the reader. Conclusion: The new microSTARii reader presents new features that improve the efficiency of the OSLD program at the IROC HQAC. New characterization is needed before final implementation is done to match the requirements of the existing OSLD system defined for the Microstar reader. Changes in uncertainty of the results has not been analized.« less

Treatment of East Coast fever: a comparison of parvaquone and buparvaquone.

PubMed

Muraguri, G R; Kiara, H K; McHardy, N

1999-11-01

Two trials were conducted to compare the efficacy of parvaquone and buparvaquone for the treatment of naturally acquired East Coast fever (ECF, Theileria parva infection) which, if untreated, is almost invariably fatal in European breeds of cattle. In the first trial 28 naive cattle were exposed in a paddock infested with ticks carrying a virulent form of the disease. Twelve were treated with each drug when they developed clinical ECF. All 24 cattle were cured. In the second study, 100 cases of ECF occurring naturally on farms in Kenya were treated, 50 with each drug. Parvaquone cured 44 (88%) buparvaquone cured 45 (90%). Intercurrent infections, predominantly anaplasmosis and bacterial pneumonia or scour, were treated specifically. It is concluded that parvaquone and buparvaquone are similarly effective in curing ECF and cure rates are maximised by accurate diagnosis and prompt treatment of both ECF and intercurrent infections.

A Novel Mach-Zehnder Interferometer Using Eccentric-Core Fiber Design for Optical Coherence Tomography.

PubMed

Xiong, Qiao; Tong, Xinglin; Deng, Chengwei; Zhang, Cui; Wang, Pengfei; Zheng, Zhiyuan; Liu, Fang

2018-05-13

A novel Mach-Zehnder interferometer using eccentric-core fiber (ECF) design for optical coherence tomography (OCT) is proposed and demonstrated. Instead of the commercial single-mode fiber (SMF), the ECF is used as one interference arm of the implementation. Because of the offset location of the eccentric core, it is sensitive to directional bending and the optical path difference (OPD) of two interference arms can be adjusted with high precision. The birefringence of ECF is calculated and experimentally measured, which demonstrates the polarization sensitivity of the ECF proposed in the paper is similar to that of SMF. Such a structure can replace the reference optical delay line to form an all-fiber passive device. A mirror is used as a sample for analyzing the ECF bending responses of the system. Besides, four pieces of overlapping glass slides as sample are experimentally measured as well.

Induction of humoral immune response to multiple recombinant rhipicephalus appendiculatus antigens and their effect on tick feeding success and pathogen transmission

USDA-ARS?s Scientific Manuscript database

Background: Rhipicephalus appendiculatus is the primary vector of Theileria parva, the etiologic agent of East Coast fever (ECF), a devastating disease of cattle in sub-Saharan Africa. We hypothesized that a vaccine targeting tick proteins that are involved in attachment and feeding might affect fee...

A systematic review of the benefit of total parenteral nutrition in the management of enterocutaneous fistulas.

PubMed

Sepehripour, S; Papagrigoriadis, S

2010-10-01

Enterocutaneous fistulas (ECFs) most commonly occur as a complication of abdominal surgery but can also occur spontaneously in inflammatory bowel disease, diverticulitis, radiation, trauma and sepsis. Although mortality and morbidity have reduced in recent years they are still a major cause of concern in patients with ECF. Nutritional support is a challenging issue in these patients and a major cause of mortality and morbidity. Total parenteral nutrition (TPN) is widely used in the management of ECF. In this review the authors examined the evidence of the use of TPN in ECF with the aim of determining the indications, benefits and outcome of this type of nutritional support in these.

DERMAL DRUG LEVELS OF ANTIBIOTIC (CEPHALEXIN) DETERMINED BY ELECTROPORATION AND TRANSCUTANEOUS SAMPLING (ETS) TECHNIQUE

PubMed Central

Sammeta, SM; Vaka, SRK; Murthy, S. Narasimha

2009-01-01

The purpose of this project was to assess the validity of a novel “Electroporation and transcutaneous sampling (ETS)” technique for sampling cephalexin from the dermal extracellular fluid (ECF). This work also investigated the plausibility of using cephalexin levels in the dermal ECF as a surrogate for the drug level in the synovial fluid. In vitro and in vivo studies were carried out using hair less rats to assess the workability of ETS. Cephalexin (20mg/kg) was administered i.v. through tail vein and the time course of drug concentration in the plasma was determined. In the same rats, cephalexin concentration in the dermal ECF was determined by ETS and microdialysis techniques. In a separate set of rats, only intraarticular microdialysis was carried out determine the time course of cephalexin concentration in synovial fluid. The drug concentration in the dermal ECF determined by ETS and microdialysis did not differ significantly from each other and so as were the pharmacokinetic parameters. The results provide validity to the ETS technique. Further, there was a good correlation (~0.9) between synovial fluid and dermal ECF levels of cephalexin indicating that dermal ECF levels could be used as a potential surrogate for cephalexin concentration in the synovial fluid. PMID:19067398

Metabolic and nutritional support of the enterocutaneous fistula patient: a three-phase approach.

PubMed

Polk, Travis M; Schwab, C William

2012-03-01

The care and outcome of enterocutaneous fistula (ECF) have improved greatly over several decades due to revolutionary advances in nutrition, along with dramatic improvements in the treatment of sepsis and the critically ill. However, as the collective experience with damage control surgery has matured, the frequent development of enteroatmospheric fistula (EAF) in the "open abdomen" patient has emerged as an even more vexing problem. Despite our best efforts, ECF and especially EAF continue to be highly morbid conditions, and sepsis and malnutrition remain the leading causes of death. Aggressive nutritional and metabolic support is the most significant predictor of outcome with ECF and EAF. Discussion of the historical advances in nutritional therapy and their impact on ECF, as well as review of the classification of ECF and EAF, provides a framework for the suggested phased strategy that specifically targets the nutritional and metabolic needs of the ECF/EAF patient. These three phases include (1) diagnosis, resuscitation, and early interval nutrition; (2) definition of fistula anatomy, drainage of collections, nutritional assessment and monitoring, and placement of feeding access; and (3) definitive nutritional management, including pharmacologic adjuncts. Early nutritional support with parenteral nutrition followed by transition to enteral nutrition is advocated, including the merits of delivery of enteral nutrition via the fistula itself, known as fistuloclysis. Aggressive nutritional therapy is necessary to reverse the catabolic state associated with ECF/EAF patients. Once established, it allows proper time, preparation, and planning for definitive management of the fistula, and in many cases provides the support for spontaneous closure.

Unchanged Erythrocyte Profile After Exposure to Cryogenic Temperatures in Elder Marathon Runners.

PubMed

Szymura, Jadwiga; Wiecek, Magdalena; Maciejczyk, Marcin; Gradek, Joanna; Kantorowicz, Malgorzata; Szygula, Zbigniew

2018-01-01

Objective: Endurance runners may experience "sports anemia" resulting from intravascular hemolysis. In addition, aging has negative impact on hematopoiesis and rheological properties of blood, and erythrocyte membranes in older people are more vulnerable to oxidative damage, which together can lead to anemia. Whole-body cryostimulation (WBCST) is increasingly used in the elderly as a method of biological regeneration of athletes or therapy and preventive treatment. That is why the aim of the study was to determine whether repeated WBCST had an effect on the erythrocyte system in master marathon runners, compared to non-training men. Methods: Ten marathon runners (men aged 55.9 ± 5.5 years, training experience 6.71 ± 5.79 years) and 10 non-training (men aged 62.0 ± 5.8 years) were subjected to a series of 24 WBCST (3 min, -130°C) performed every other day. Erythrocyte levels, interleukin-3 (IL-3), erythropoietin (EPO), haptoglobin, bilirubin, and extracellular hemoglobin (HGB ecf ) concentrations were determined in the blood before and after 12, 24 WBCST, as well as 7 days after their completion. Results: The concentrations of EPO and IL-3 were significantly increased 7 days after the completion of WBCST in both groups ( P < 0.05). The erythrocyte content and indicators, the bilirubin, haptoglobin, and HGB ecf levels in each group did not change as a result of WBCST. In order to document hemolytic changes and/or factors affecting the severity of erythropoiesis, correlations between growth erythropoietic factors, erythrocyte and hemolytic factors as well as mutual correlations between hemolytic indexes were calculated. There was a positive correlation ( P < 0.05) between the EPO and IL-3, bilirubin, mean corpuscular hemoglobin, and red blood cell distribution width - standard deviation. There was also a positive correlation between the concentrations of bilirubin and HGB ecf , and a negative correlation between haptoglobin and HGB ecf as well as bilirubin concentrations. Conclusion: WBCST treatments, repeated every other day, do not cause hemolytic changes in elder men with high or low physical activity. But also, they are a procedure that does not increase the level of erythrocytes or their hemoglobinization. In athletes, it is not a form of doping. The positive correlation between EPO and bilirubin may be indicative of, for example, the mutual antioxidative effect of these factors.

Use of the ecf1 gene to detect Shiga toxin-producing Escherichia coli in beef samples.

PubMed

Livezey, Kristin W; Groschel, Bettina; Becker, Michael M

2015-04-01

Escherichia coli O157:H7 and six serovars (O26, O103, O121, O111, O145, and O45) are frequently implicated in severe clinical illness worldwide. Standard testing methods using stx, eae, and O serogroup-specific gene sequences for detecting the top six non-O157 STEC bear the disadvantage that these genes may reside, independently, in different nonpathogenic organisms, leading to false-positive results. The ecf operon has previously been identified in the large enterohemolysin-encoding plasmid of eae-positive Shiga toxin-producing E. coli (STEC). Here, we explored the utility of the ecf operon as a single marker to detect eae-positive STEC from pure broth and primary meat enrichments. Analysis of 501 E. coli isolates demonstrated a strong correlation (99.6%) between the presence of the ecf1 gene and the combined presence of stx, eae, and ehxA genes. Two large studies were carried out to determine the utility of an ecf1 detection assay to detect non-O157 STEC strains in enriched meat samples in comparison to the results using the U. S. Department of Agriculture Food Safety and Inspection Service (FSIS) method that detects stx and eae genes. In ground beef samples (n = 1,065), the top six non-O157 STEC were detected in 4.0% of samples by an ecf1 detection assay and in 5.0% of samples by the stx- and eae-based method. In contrast, in beef samples composed largely of trim (n = 1,097), the top six non-O157 STEC were detected at 1.1% by both methods. Estimation of false-positive rates among the top six non-O157 STEC revealed a lower rate using the ecf1 detection method (0.5%) than using the eae and stx screening method (1.1%). Additionally, the ecf1 detection assay detected STEC strains associated with severe illness that are not included in the FSIS regulatory definition of adulterant STEC.

Induction of a stable sigma factor SigR by translation-inhibiting antibiotics confers resistance to antibiotics.

PubMed

Yoo, Ji-Sun; Oh, Gyeong-Seok; Ryoo, Sungweon; Roe, Jung-Hye

2016-06-27

Antibiotic-producing streptomycetes are rich sources of resistance mechanisms against endogenous and exogenous antibiotics. An ECF sigma factor σ(R) (SigR) is known to govern the thiol-oxidative stress response in Streptomyces coelicolor. Amplification of this response is achieved by producing an unstable isoform of σ(R) called σ(R'). In this work, we present evidence that antibiotics induce the SigR regulon via a redox-independent pathway, leading to antibiotic resistance. The translation-inhibiting antibiotics enhanced the synthesis of stable σ(R), eliciting a prolonged response. WblC/WhiB7, a WhiB-like DNA-binding protein, is responsible for inducing sigRp1 transcripts encoding the stable σ(R). The amount of WblC protein and its binding to the sigRp1 promoter in vivo increased upon antibiotic treatment. A similar phenomenon appears to exist in Mycobacterium tuberculosis as well. These findings reveal a novel antibiotic-induced resistance mechanism conserved among actinomycetes, and also give an explicit example of overlap in cellular damage and defense mechanisms between thiol-oxidative and anti- translational stresses.

Induction of a stable sigma factor SigR by translation-inhibiting antibiotics confers resistance to antibiotics

PubMed Central

Yoo, Ji-Sun; Oh, Gyeong-Seok; Ryoo, Sungweon; Roe, Jung-Hye

2016-01-01

Antibiotic-producing streptomycetes are rich sources of resistance mechanisms against endogenous and exogenous antibiotics. An ECF sigma factor σR (SigR) is known to govern the thiol-oxidative stress response in Streptomyces coelicolor. Amplification of this response is achieved by producing an unstable isoform of σR called σR′. In this work, we present evidence that antibiotics induce the SigR regulon via a redox-independent pathway, leading to antibiotic resistance. The translation-inhibiting antibiotics enhanced the synthesis of stable σR, eliciting a prolonged response. WblC/WhiB7, a WhiB-like DNA-binding protein, is responsible for inducing sigRp1 transcripts encoding the stable σR. The amount of WblC protein and its binding to the sigRp1 promoter in vivo increased upon antibiotic treatment. A similar phenomenon appears to exist in Mycobacterium tuberculosis as well. These findings reveal a novel antibiotic-induced resistance mechanism conserved among actinomycetes, and also give an explicit example of overlap in cellular damage and defense mechanisms between thiol-oxidative and anti- translational stresses. PMID:27346454

A primary estimation of PCDD/Fs release reduction from non-wood pulp and paper industry in China based on the investigation of pulp bleaching with chlorine converting to chlorine dioxide.

PubMed

Xiao, Qingcong; Song, Xiaoqian; Li, Wenchao; Zhang, Yuanna; Wang, Hongchen

2017-10-01

Chlorine bleaching technology (C process, CEH process, H process and theirs combination), which was identified as a primary formation source of PCDD/Fs, is still widely used by the vast majority of Chinese non-wood pulp and paper mills (non-wood PMs). The purpose of this study was to provide information and data support for further eliminating dioxin for non-wood PMs in China, and especially to evaluate the PCDD/Fs release reduction for those mills converting their pulp bleaching processes from CEH to ECF. The PCDD/Fs concentrations of the bleached pulp and bleaching wastewater with ECF bleaching were in the ranges of 0.13-0.8 ng TEQ kg -1 , and 0.15-1.9 pg TEQ L -1 , respectively, which were far lower than those with CEH process, indicating that the ECF process is an effective alternative bleaching technology to replace CEH in Chinese non-wood PMs to reduce dioxin release. The release factor via flue gas of the alkali recovery boiler in Chinese non-wood PMs was first reported to be 0.092 μg TEQ Ad t -1 in this study. On the assumption that pulp bleaching processes of all Chinese non-wood PMs were converted from CEH to ECF, the annual release of PCDD/Fs via the bleaching wastewater and bleached pulp would be reduced by 79.1%, with a total of 1.60 g TEQ. Copyright © 2017 Elsevier Ltd. All rights reserved.

Surface Plasmon Scattering in Exposed Core Optical Fiber for Enhanced Resolution Refractive Index Sensing.

PubMed

Klantsataya, Elizaveta; François, Alexandre; Ebendorff-Heidepriem, Heike; Hoffmann, Peter; Monro, Tanya M

2015-09-29

Refractometric sensors based on optical excitation of surface plasmons on the side of an optical fiber is an established sensing architecture that has enabled laboratory demonstrations of cost effective portable devices for biological and chemical applications. Here we report a Surface Plasmon Resonance (SPR) configuration realized in an Exposed Core Microstructured Optical Fiber (ECF) capable of optimizing both sensitivity and resolution. To the best of our knowledge, this is the first demonstration of fabrication of a rough metal coating suitable for spectral interrogation of scattered plasmonic wave using chemical electroless plating technique on a 10 μm diameter exposed core of the ECF. Performance of the sensor in terms of its refractive index sensitivity and full width at half maximum (FWHM) of SPR response is compared to that achieved with an unstructured bare core fiber with 140 μm core diameter. The experimental improvement in FWHM, and therefore the detection limit, is found to be a factor of two (75 nm for ECF in comparison to 150 nm for the large core fiber). Refractive index sensitivity of 1800 nm/RIU was achieved for both fibers in the sensing range of aqueous environment (1.33-1.37) suitable for biosensing applications.

An extracytoplasmic function sigma factor cotranscribed with its cognate anti-sigma factor confers tolerance to NaCl, ethanol and methylene blue in Azospirillum brasilense Sp7.

PubMed

Mishra, Mukti Nath; Kumar, Santosh; Gupta, Namrata; Kaur, Simarjot; Gupta, Ankush; Tripathi, Anil K

2011-04-01

Azospirillum brasilense, a plant-growth-promoting rhizobacterium, is exposed to changes in its abiotic environment, including fluctuations in temperature, salinity, osmolarity, oxygen concentration and nutrient concentration, in the rhizosphere and in the soil. Since extra-cytoplasmic function (ECF) sigma factors play an important role in stress adaptation, we analysed the role of ECF sigma factor (also known as RpoE or σ(E)) in abiotic stress tolerance in A. brasilense. An in-frame rpoE deletion mutant of A. brasilense Sp7 was carotenoidless and slow-growing, and was sensitive to salt, ethanol and methylene blue stress. Expression of rpoE in the rpoE deletion mutant complemented the defects in growth, carotenoid biosynthesis and sensitivity to different stresses. Based on data from reverse transcriptase-PCR, a two-hybrid assay and a pull-down assay, we present evidence that rpoE is cotranscribed with chrR and the proteins synthesized from these two overlapping genes interact with each other. Identification of the transcription start site by 5' rapid amplification of cDNA ends showed that the rpoE-chrR operon was transcribed by two promoters. The proximal promoter was less active than the distal promoter, whose consensus sequence was characteristic of RpoE-dependent promoters found in alphaproteobacteria. Whereas the proximal promoter was RpoE-independent and constitutively expressed, the distal promoter was RpoE-dependent and strongly induced in response to stationary phase and elevated levels of ethanol, salt, heat and methylene blue. This study shows the involvement of RpoE in controlling carotenoid synthesis as well as in tolerance to some abiotic stresses in A. brasilense, which might be critical in the adaptation, survival and proliferation of this rhizobacterium in the soil and rhizosphere under stressful conditions.

Damage indication in smart structures using modal effective electromechanical coupling coefficients

NASA Astrophysics Data System (ADS)

Al-Ajmi, M. A.; Benjeddou, A.

2008-06-01

This work explores the use, in structural health monitoring, of the so-called modal effective electromechanical coupling coefficient (EMCC) as a damage indicator for structures with failures such as cracks. For this purpose, a discrete layered finite element (FE) model for smart beams is proposed and applied to short-circuit (SC) and open-circuit (OC) modal analyses of healthy and damaged (cracked) cantilever beams with symmetrically surface-bonded piezoelectric patches. Focus is made here on enhancing the electrical behavior modeling by introducing a quadratic bubble function in the electric potential through-the-thickness approximation. Therefore, the corresponding higher-order potential (HOP) degree of freedom is condensed at the ply level, leading to a passive stiffening effect (SE) similar to the so-called higher-order induced potential (HIP); then the physical equipotential (EP) electrode effect, often neglected in the piezoelectric FE literature, is here implemented after the electrodes' FE assembly. After its validation against available analytical and experimental results, the proposed piezoelectric FE is used for parametric analyses of SC-based and OC-based EMCC change factors (ECFs) and frequency change factors (FCFs) in terms of the crack depth and position ratios. It was found that the EP effect was more influential on the ECF than the SE. However, for the FCFs, the EP effect was influential only when it is defined from the OC frequencies. Finally, the ECFs were found to be higher than the FCFs, in particular for higher modes.

Effect of different East Coast fever control strategies on disease incidence in traditionally managed Sanga cattle in Central Province of Zambia.

PubMed

Minjauw, B; Otte, M J; James, A D; de Castro, J J; Sinyangwe, P

1998-05-01

A clinical trial, including five East Coast fever (ECF) control strategies (involving tick control and/or immunisation by infection-and-treatment) in five different groups of traditionally managed Sanga cattle, was conducted in Central Province of Zambia over 2.5 years between 1992 and 1995. Two groups were kept under intensive tick control by weekly acaricide treatment by hand spray; (one immunised and one non-immunised), two groups were under no tick control (one immunised and one non-immunised), and a fifth, immunised group was maintained under strategic tick control (18 sprays yr-1). ECF-specific mortality was highest in the non-immunised and non-treated group, while no difference in ECF-specific mortality could be observed between animals treated for ECF by immunisation or by tick control. Acaricide treatment and/or immunisation reduced the risk of clinical ECF by 92%. The results of an artificial challenge experiment at the end of the field trial indicated that about 60% of the animals in the control group had become infected with Theileria parva without showing clinical signs. ECF incidence in non-vaccinated cattle markedly declined six months after immunisation--suggesting that the carrier state induced by immunisation did not lead to a persistent high incidence, and might accelerate the progress to endemicity.

A three-dimensional model of the atmospheric chemistry of E and Z-CF3CH=CHCl (HCFO-1233(zd) (E/Z))

NASA Astrophysics Data System (ADS)

Sulbaek Andersen, Mads P.; Schmidt, Johan A.; Volkova, Aleksandra; Wuebbles, Donald J.

2018-04-01

Using a 3-dimensional global atmospheric chemistry and transport model we investigated the atmospheric degradation of HCFO-1233zd(E), E-CF3CH=CHCl, a commercially important, new hydrofluorocarbon replacement compound. Atmospheric degradation of E-CF3CH=CHCl is initiated by reaction with OH radicals, which leads to several chemical oxidation products. Dissemination of these oxidation products to the environment is of concern due to the possible formation of trifluoroacetic acid (TFA) as a degradation product. The model indicates that the average global yield of TFA from atmospheric processing of E-CF3CH=CHCl is approximately 2%. The annually averaged atmospheric lifetime of E-CF3CH=CHCl was found to be approximately 36 days (12 days for Z-CF3CH=CHCl). As E-CF3CH=CHCl is short lived, by far the majority of its Cl atoms will be released and deposited in the lower atmosphere, and the impact on stratospheric ozone is insignificant. An Ozone Depletion Potential of 0.00030 was determined. The Photochemical Ozone Creation Potential was evaluated and a value of 3.6 determined. Finally, we derive a Global Warming Potential for E-CF3CH=CHCl for a 100 year time horizon of <5. For comparison, data for the stereoismeric analogue, Z-CF3CH=CHCl, was also obtained from the model.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Umino, Satoru; Takahashi, Hideaki, E-mail: hideaki@m.tohoku.ac.jp; Morita, Akihiro

In a recent work, we developed a method [H. Takahashi et al., J. Chem. Phys. 143, 084104 (2015)] referred to as exchange-core function (ECF) approach, to compute exchange repulsion E{sub ex} between solute and solvent in the framework of the quantum mechanical (QM)/molecular mechanical (MM) method. The ECF, represented with a Slater function, plays an essential role in determining E{sub ex} on the basis of the overlap model. In the work of Takahashi et al. [J. Chem. Phys. 143, 084104 (2015)], it was demonstrated that our approach is successful in computing the hydrogen bond energies of minimal QM/MM systems includingmore » a cationic QM solute. We provide in this paper the extension of the ECF approach to the free energy calculation in condensed phase QM/MM systems by combining the ECF and the QM/MM-ER approach [H. Takahashi et al., J. Chem. Phys. 121, 3989 (2004)]. By virtue of the theory of solutions in energy representation, the free energy contribution δμ{sub ex} from the exchange repulsion was naturally formulated. We found that the ECF approach in combination with QM/MM-ER gives a substantial improvement on the calculation of the hydration free energy of a hydronium ion. This can be attributed to the fact that the ECF reasonably realizes the contraction of the electron density of the cation due to the deficit of an electron.« less

An Enhanced Convective Forecast (ECF) for the New York TRACON Area

NASA Technical Reports Server (NTRS)

Wheeler, Mark; Stobie, James; Gillen, Robert; Jedlovec, Gary; Sims, Danny

2008-01-01

In an effort to relieve summer-time congestion in the NY Terminal Radar Approach Control (TRACON) area, the FAA is testing an enhanced convective forecast (ECF) product. The test began in June 2008 and is scheduled to run through early September. The ECF is updated every two hours, right before the Air Traffic Control System Command Center (ATCSCC) national planning telcon. It is intended to be used by traffic managers throughout the National Airspace System (NAS) and airlines dispatchers to supplement information from the Collaborative Convective Forecast Product (CCFP) and the Corridor Integrated Weather System (CIWS). The ECF begins where the current CIWS forecast ends at 2 hours and extends out to 12 hours. Unlike the CCFP it is a detailed deterministic forecast with no aerial coverage limits. It is created by an ENSCO forecaster using a variety of guidance products including, the Weather Research and Forecast (WRF) model. This is the same version of the WRF that ENSCO runs over the Florida peninsula in support of launch operations at the Kennedy Space Center. For this project, the WRF model domain has been shifted to the Northeastern US. Several products from the NASA SPoRT group are also used by the ENSCO forecaster. In this paper we will provide examples of the ECF products and discuss individual cases of traffic management actions using ECF guidance.

Treatment of East Coast fever of cattle with a combination of parvaquone and frusemide.

PubMed

Musoke, R Azuba; Tweyongyere, R; Bizimenyera, E; Waiswa, C; Mugisha, A; Biryomumaisho, S; McHardy, N

2004-04-01

Pulmonary oedema is a common sign of East Coast fever (ECF, Theileria parva infection) of cattle. A trial was conducted on farms in Uganda to compare a product containing both the antitheilerial compound parvaquone and the diuretic compound frusemide with one containing only parvaquone, in the treatment of ECF. The trial involved 40 clinical cases of ECF, some of them complicated by other infections, in cattle of all ages and on several farms. Confirmed cases were treated with either parvaquone+frusemide (P+F) or parvaquone alone (P). Survival after treatment with P+F was 77% compared with 71% with P. Five of the 10 fatalities were complicated cases. The cure rate for severe but uncomplicated ECF was 89% with P+F and 40% with P. Pulmonary signs were resolved within 24-48 h after treatment with P+F and clinical recovery was noticeably more rapid than with P. The antiparasitic effect of the two treatments was similar. P+F could be particularly useful when reporting, diagnosis or laboratory confirmation of ECF is delayed, because advanced cases are more likely to be encountered under these circumstances.

Enhancement of Chlorella vulgaris harvesting via the electro-coagulation-flotation (ECF) method.

PubMed

Wong, Y K; Ho, Y H; Leung, H M; Ho, K C; Yau, Y H; Yung, K K L

2017-04-01

This article explores the potential of using an electro-coagulation-flotation (ECF) harvester to allow flotation of microalgae cells for surface harvesting. A response surface methodology (RSM) model was used to optimize ECF harvesting by adjusting electrode plate material, electrode plate number, charge of the electrodes, electrolyte concentration, and pH value of the culture solution. The result revealed that three aluminum electrode plates (one anode and two cathodes), brine solution (8 g/L), and acidity (pH = 4) of culture solution (optimized ECF harvester) The highest flocculant concentration was measured at 2966 mg/L after 60 min and showed a 79.8 % increase of flocculation concentration. Such results can provide a basis for designing a large-scale microalgae harvester for commercial use in the future.

General radiographic attributes of optically stimulated luminescence dosimeters: A basic insight

NASA Astrophysics Data System (ADS)

Musa, Y.; Hashim, S.; Ghoshal, S. K.; Bradley, D. A.; Ahmad, N. E.; Karim, M. K. A.; Hashim, A.; Kadir, A. B. A.

2018-06-01

We report the ubiquitous radiographic characteristics of optically stimulated luminescence dosimeters (OSLD) so called nanoDot OSLDs (Landauer Inc., Glendwood, IL). The X-ray irradiations were performed in free air ambiance to inspect the repeatability, the reproducibility, the signal depletion, the element correction factors (ECFs), the dose response and the energy dependence. Repeatability of multiple readouts after single irradiation to 10 mGy revealed a coefficient of variation below 3%, while the reproducibility in repeated irradiation-readout-annealing cycles was above 2%. The OSL signal depletion for three nanoDots with simultaneous irradiation to 20 mGy and sequential readouts of 25 times displayed a consistent signal reduction ≈0.5% per readout with R2 values over 0.98. ECFs for individual OSLDs were varied from 0.97 to 1.03. In the entire dose range under 80 kV, a good linearity with an R2 exceeding 0.99 was achieved. Besides, the percentage difference between OSLD and ion-chamber dose was less than 5%, which was superior to TLD. The X-ray photon irradiated energy response factors (between 0.76 and 1.12) in the range of 40-150 kV (26.1-61.2 keV) exhibited significant energy dependence. Indeed, the nanoDot OSLDs disclosed good repeatability, reproducibility and linearity. The OSLDs measured doses were closer to ion-chamber doses than that of TLD. It can be further improved up to ≈3% by applying the individual dosimeter ECF. On top, the energy dependent uncertainties can be minimized using the energy correction factors. It is established that the studied nanoDot OSLDs are prospective for measuring entrance dose in general radiographic practices.

Antibodies to in silico selected GPI-anchored Theileria parva proteins neutralize sporozoite infection in vitro.

PubMed

Nyagwange, James; Nene, Vishvanath; Mwalimu, Stephen; Henson, Sonal; Steinaa, Lucilla; Nzau, Benjamin; Tijhaar, Edwin; Pelle, Roger

2018-05-01

East Coast fever (ECF) caused by Theileria parva kills cattle in East, Central and Southern Africa leading to significant economic losses. Vaccination is used as a control strategy against ECF and is presently dependent on deliberate infection with live sporozoites and simultaneous treatment with a long-acting oxytetracycline. Although effective, this method has serious limitations; the immunity is parasite strain specific and immunized cattle can become life-long asymptomatic carriers of the parasite, posing risk for the spread of the disease. In efforts to develop a subunit vaccine, the role of antibodies in the neutralization of T. parva sporozoites infection of host cells has been investigated and a circumsporozoite protein, p67, is able to induce such neutralizing antibodies. However, the p67 protein only protects a proportion of immunized cattle against T. parva challenge and such protection might be improved by inclusion of additional parasite antigens that neutralize sporozoite infection. In an attempt to identify such antigens, we searched the re-annotated T. parva genome for genes predicted to contain GPI anchor signals, since they are likely to be located on the cell surface, and expressed fragments of six of the selected genes in E. coli. The recombinant proteins were used to raise antisera in mice. Antisera to two proteins, TpMuguga_01g00876 and TpMuguga_01g00939, neutralized sporozoite infectivity to a high degree, while antisera to two additional proteins, TpMuguga_01g00095 and TpMuguga_04g00437, exhibited moderate neutralizing capacity. We conclude that these four antigens are potential vaccine candidates, which should be evaluated further in cattle. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Theileria parva epidemics: a case study in eastern Zambia.

PubMed

Billiouw, M; Vercruysse, J; Marcotty, T; Speybroeck, N; Chaka, G; Berkvens, D

2002-07-29

This paper presents the results of the follow-up of three sentinel herds between 1994 and 2000 during an East Coast fever (ECF) epidemic in eastern Zambia. The animals of the sentinel herds were closely monitored clinically and serologically together with detailed Rhipicephalus appendiculatus counts. Peaks of disease incidence occurred in the rainy season (December-February) and the dry months of May-July with nymph-to-adult tick transmission dominating the infection dynamics. A second wave of adult R. appendiculatus at the start of the dry season is essential for the occurrence of a full-blown epidemic while the size of the susceptible cattle population acts as a most important limiting factor. The majority of adult cattle of the sentinel herds became infected less than 2 years after the introduction of the disease. The median age at first contact for calves born towards the end of the study (1999) was about 6 months. The case-fatality ratio (including sub-clinical cases) is estimated at 60%. It is argued that part of the so-called 'natural mortality' is actually due to ECF and that ECF occurrence and mortality are systematically underestimated. The direct financial cost of the epidemic, based on loss of animals and cost of treatment only and calculated over 4 years running, is estimated at about 6 US dollars per year per animal at risk. The value of the traditional seroprevalence survey as a tool for monitoring ECF epidemiology is put in question and the prevalence of maternal antibodies in new-born calves, reflecting the immune status of the dam population, is introduced as an alternative. It is demonstrated that an efficient immunisation campaign should concentrate its efforts in the period of low adult R. appendiculatus abundance (July-October).

Uptake of Biotin by Chlamydia Spp. through the Use of a Bacterial Transporter (BioY) and a Host-Cell Transporter (SMVT)

PubMed Central

Fisher, Derek J.; Fernández, Reinaldo E.; Adams, Nancy E.; Maurelli, Anthony T.

2012-01-01

Chlamydia spp. are obligate intracellular Gram-negative bacterial pathogens that cause disease in humans and animals. Minor variations in metabolic capacity between species have been causally linked to host and tissue tropisms. Analysis of the highly conserved genomes of Chlamydia spp. reveals divergence in the metabolism of the essential vitamin biotin with genes for either synthesis (bioF_2ADB) and/or transport (bioY). Streptavidin blotting confirmed the presence of a single biotinylated protein in Chlamydia. As a first step in unraveling the need for divergent biotin acquisition strategies, we examined BioY (CTL0613) from C. trachomatis 434/Bu which is annotated as an S component of the type II energy coupling-factor transporters (ECF). Type II ECFs are typically composed of a transport specific component (S) and a chromosomally unlinked energy module (AT). Intriguingly, Chlamydia lack recognizable AT modules. Using 3H-biotin and recombinant E. coli expressing CTL0613, we demonstrated that biotin was transported with high affinity (a property of Type II ECFs previously shown to require an AT module) and capacity (apparent K(m) of 3.35 nM and V(max) of 55.1 pmol×min−1×mg−1). Since Chlamydia reside in a host derived membrane vacuole, termed an inclusion, we also sought a mechanism for transport of biotin from the cell cytoplasm into the inclusion vacuole. Immunofluorescence microscopy revealed that the mammalian sodium multivitamin transporter (SMVT), which transports lipoic acid, biotin, and pantothenic acid into cells, localizes to the inclusion. Since Chlamydia also are auxotrophic for lipoic and pantothenic acids, SMVT may be subverted by Chlamydia to move multiple essential compounds into the inclusion where BioY and another transporter(s) would be present to facilitate transport into the bacterium. Collectively, our data validates the first BioY from a pathogenic organism and describes a two-step mechanism by which Chlamydia transport biotin from the host cell into the bacterial cytoplasm. PMID:23029384

Space Station requirements for in-flight exercise countermeasures

NASA Technical Reports Server (NTRS)

Hayes, Judith C.; Harris, Bernard A.

1990-01-01

In an effort to retard the deleterious effects of space adaptation, NASA has defined requirements for an Exercise Countermeasure Facility (ECF) within the Space Station Crew Health Care System (CHeCS). The application of exercise as a countermeasure to spaceflight-induced deconditioning has been utilized in the past by both the United States and the Soviet space programs. The ECF will provide exercise hardware, physiological monitoring capabilities, and an interactive motivational display system. ECF operations and data will be coupled through the Space Station Freedom Data Management System for monitoring of inflight training and testing from ground control, thus allowing for real-time evaluation of crewmember performance and modification of exercise prescriptions. Finally, the objective of the ECF is to monitor and control the exercise of crewmembers for the maintenance of an operational level of fitness to ensure mission success.

Alternative sigma factor RpoN and its modulation protein YhbH are indispensable for Erwinia amylovora virulence.

PubMed

Ancona, Veronica; Li, Wenting; Zhao, Youfu

2014-01-01

In Erwinia amylovora, ECF (extracytoplasmic functions) alternative sigma factor HrpL regulates the transcription of hrp (hypersensitive response and pathogenicity)-type III secretion system (T3SS) genes by binding to a consensus sequence known as the hrp box in hrp gene promoters. In turn, the expression of hrpL has been proposed to be positively controlled by alternative sigma factor 54 (σ(54)) (RpoN) and HrpS, a member of the σ(54) enhancer-binding proteins (EBPs). However, the function of RpoN has not been characterized genetically in E. amylovora. In this study, we investigated the role of RpoN, a nitrogen limitation sigma factor, and its modulation protein YhbH, a novel ribosome-associated protein, in E. amylovora virulence. Our results showed that mutations in hrpS, hrpL, rpoN and yhbH, but not yfiA and rmf3, resulted in a nonpathogenic phenotype on immature pear fruits and apple shoots. Consistently, the expression of T3SS genes, including hrpL, dspE, hrpN and hrpA, was barely detected in hrpS, hrpL, rpoN and yhbH mutants. These mutants were also not capable of eliciting a hypersensitive response (HR) on tobacco; however, the overexpression of hrpL using an inducible promoter rescued the HR-eliciting abilities of these mutants. These results suggest that a sigma factor cascade exists in the regulatory networks of E. amylovora and regulates important virulence factors. On the basis of this study and previously reported data, a model is proposed for the regulation of T3SS in E. amylovora. © 2013 BSPP AND JOHN WILEY & SONS LTD.

Comparative studies of the efficacy of parvaquone and parvaquone-plus-frusemide in the treatment of Theileria parva infection (East Coast fever) in cattle.

PubMed

Mbwambo, H A; Sudi, F F; Mkonyi, P A; Mfinanga, J M; Mella, E S; Ngovi, C J

2002-09-26

Comparative studies of the efficacy of parvaquone (Parvexon) and parvaquone-plus-frusemide (Fruvexon) Bimeda Chemicals, Ireland, were done on 60 naturally infected cases of East Coast fever (ECF; Theileria parva infection in cattle). Small-scale dairy farmers in the peri-urban of Dar Es Salaam city reported ECF-suspected cases from March to mid-October 2001 and were treated with the two drugs alternately, as were diagnosed positive for ECF. Four sub-groups of 15 cattle each (early stage, 15; advanced stage, 15) were treated with parvaquone and parvaquone-plus-frusemide. Twenty-eight out of 30 (93.3%) cattle treated with parvaquone-plus-frusemide were cured, so do 24 out of 30 (80.0%) cattle treated with parvaquone without frusemide. Early diagnosis and prompt management of pulmonary signs, which accounted for 30.0% of total ECF cases is advised in order to improve cure rates. Unlike parvaquone without frusemide (Parvexon), parvaquone-plus-frusemide (Fruvexon) proved useful in the management of pulmonary signs, hence, a drug of choice in the treatment of ECF cases that are accompanied by or are likely to manifest pulmonary signs.

77 FR 3380 - Airworthiness Directives; Eurocopter France (ECF) Model AS350B, B1, B2, B3, BA, and D; and AS355E...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-01-24

... Airworthiness Directives; Eurocopter France (ECF) Model AS350B, B1, B2, B3, BA, and D; and AS355E, F, F1, F2... Register as of February 8, 2012. We must receive comments on this AD by March 26, 2012. ADDRESSES: You may..., 2009, specifying MOD 073330 for the Model SA350B, B1, B2, B3, BA, BB, D, and L1 helicopters. ECF has...

The European HST Science Data Archive. [and Data Management Facility (DMF)

NASA Technical Reports Server (NTRS)

Pasian, F.; Pirenne, B.; Albrecht, R.; Russo, G.

1993-01-01

The paper describes the European HST Science Data Archive. Particular attention is given to the flow from the HST spacecraft to the Science Data Archive at the Space Telescope European Coordinating Facility (ST-ECF); the archiving system at the ST-ECF, including the hardware and software system structure; the operations at the ST-ECF and differences with the Data Management Facility; and the current developments. A diagram of the logical structure and data flow of the system managing the European HST Science Data Archive is included.

Prevalence and risk factors associated with Theileria parva infection in cattle in three regions of Tanzania.

PubMed

Kerario, Isack I; Simuunza, Martin C; Chenyambuga, Sebastian W; Koski, Marja; Hwang, Seong-Gu; Muleya, Walter

2017-12-01

Ticks and tickborne diseases (TBDs) are serious constraints to cattle production in Tanzania and other tropical and subtropical countries. Among the TBDs, East Coast fever (ECF) is the most important as it causes significant economic losses to the cattle industry in Tanzania. However, control of ECF in Tanzania has continued to be a challenge due to inadequate epidemiological information. The main objective of this study was to determine the epidemiological situation of Theileria parva infections in cattle kept under pastoral and agro-pastoral farming systems in Mara, Singida, and Mbeya regions of Tanzania. Blood samples were collected from 648 cattle in the three regions. Genomic DNA was extracted and amplified in a polymerase chain reaction (PCR) using T. parva-specific primers targeting the 104-kD antigen (P104) gene. In addition, information was collected on the possible risk factors of T. parva infection (animal age, region, animal sex, tick burden, tick control method, and frequency of acaricide application). The prevalence of T. parva across the three regions was 14.2%. There was variation in prevalence among the three regions with Mara (21.8%) having a significantly higher (p = 0.001) prevalence than the other regions. Moreover, Mbeya exhibited relatively lower prevalence (7.4%) compared to the other regions. Factors found to be significantly associated with an animal being PCR positive for T. parva were region (p = 0.001) and tick burden (p = 0.003). Other factors were not found to be significant predictors of being PCR positive for T. parva. The present study showed high variation in tick burden and T. parva prevalence across the regions. Therefore, different strategic planning and cost-effective control measures for ticks and T. parva infection should be implemented region by region in order to reduce losses caused by ticks and ECF in the study area.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Miller, Guthrie; Klumpp, John A.; Melo, Dunstana

Here, the pharmacokinetic equations of Pierson et al. describing the behavior of bromide in rat provide a general approach to the modeling of extracellular fluid (ECF). The movement of material into ECF spaces is rapid and is completely characterized by tissue volumes and vascular flow rates to and from a tissue, the volumes of the tissue, and the ECF associated with the tissue. Early-time measurements are needed to characterize ECF. Measurements of DTPA disappearance from plasma by Wedeking et al. are discussed as an example of such measurements. In any biokinetic model, the fastest transfer rates are not determinable withmore » the usual datasets, and if determined empirically, these rates will have very large and highly correlated uncertainties, so particular values of these rates, even though the model fits the available data, are not significant. A pharmacokinetic front-end provides values for these fast rates. An example of such a front-end for a 200–g rat is given.« less

Ependymin, a brain extracellular glycoprotein, and CNS plasticity.

PubMed

Shashoua, V E

1991-01-01

Ependymin, a glycoprotein of the brain ECF, has been implicated in the neurochemistry of memory and neuronal regeneration. Three behavioral experiments (swimming with a float, avoidance conditioning, and classical conditioning) in the goldfish and one in the mouse (T-maze learning) indicate that ependymin has a role in the synaptic changes that take place in the consolidation step of memory formation and the activity-dependent phase of sharpening of goldfish retinotectal connections during neuronal regeneration. The ECF concentration of the protein was found to decrease after the goldfish learned to associate a light stimulus (CS) with the subsequent arrival of a shock (US): paired CS-US gave changes whereas an unpaired presentation of CS-US gave no changes relative to the unstimulated controls. Ependymin is present in ECF as a mixture of three disulfide-linked dimers of two acidic (alpha and beta) polypeptide chains (37 kDa and 31 kDa). Upon removal of its N-linked glycan fragment by N-glycosidase F, the beta chain yields gamma-ependymin (26 kDa). Determinations of the amino acid sequence of gamma-ependymin indicate that it is a unique protein with no long sequence homologies to any known polypeptide. There are, however, small segments (5-7 amino acids long) with homologies to fibronectin, laminin, and tubulin. Ependymin has the capacity to polymerize into FIP (after activation by phosphorylation) in response to events that deplete ECF calcium. FIP is insoluble in 2% SDS in 6 M urea, 10 mM Ca2+Ac2, 100% acetic acid, chloroform/methanol (2/1), saturated KCNS, and even 100% trifluoroacetic acid. FIP was found to be present in goldfish brain and to be formed as a labeled product in vivo. Ependymin's FIP-forming property was used to propose a molecular hypothesis for generating synaptic changes in response to local extracellular depletions of calcium at sites of "associating inputs." The model assumes that, following NMDA receptor stimulation, the translocated PKC that is generated activates extracellular ependymin by converting it to its phosphorylated form using presynaptically released ATP. The hypothesis was tested in studies of LTP of rat hippocampal slices at CA1. After LTP, new sites that stained with antisera to ependymin, visible at 100x, were obtained in its potentiated radiatum in the CA1 region but not in the unpotentiated CA3. Electron microscopic studies showed that the horseradish peroxidase reaction product obtained was localized at synaptic clefts and postsynaptic regions. The results suggest that FIP may be formed at extracellular and postsynaptic loci where multiple associating inputs interact at CA1.

Data Sharing Between Providers and Quality Initiatives Eliminate Unnecessary Nursing Home Admissions.

PubMed

Charles, Ryan J; Singal, Bonita M; Urquhart, Andrew G; Masini, Michael A; Hallstrom, Brian R

2017-05-01

The Michigan Arthroplasty Registry Collaborative Quality Initiative (MARCQI) has monitored discharge disposition, after total hip and knee arthroplasties, since inception in 2012 and found the standardized risk of extended care facility (ECF) placement to be highly variable between hospitals. The variation in standardized risks of ECF placement among MARCQI member sites was reported to the collaborative. At the May 2, 2014 quarterly meeting, a quality initiative was started, emphasizing the wide variability between hospitals, the contribution of hospital and surgeon to that variability using median odds ratios, and the need for outlier hospitals to initiate quality improvement (QI) processes. Patients from 29 hospitals that were members of MARCQI before the intervention were included in this analysis. We compared standardized risks before and after the intervention in the entire cohort, and for 3 hospitals that implemented institution-specific QI projects. We report changes in ECF placement, length of stay, emergency room visits, and readmissions over time. This study includes 31,347 patients before and 20,879 patients after the implementation of the quality initiative. The range in standardized risk dropped from 9.4%-46.1% to 9.4%-32.4% and the average dropped from 23.0% to 19.6%. Three outlier hospitals decreased their absolute risk of ECF placement by 12.2%, 8.9%, and 12.4% after QI, without increases in adverse outcomes. Discharge to ECF after primary hip and knee arthroplasties is highly variable and influenced by hospital and surgeon practices. Hospital-level QI measures can decrease ECF admissions. Copyright © 2016 Elsevier Inc. All rights reserved.

AlgU controls expression of virulence genes in Pseudomonas syringae pv. tomato DC3000

USDA-ARS?s Scientific Manuscript database

Plant pathogenic bacteria are able to integrate information about their environment and adjust gene expression to provide adaptive functions. AlgU, an ECF sigma factor encoded by Pseudomonas syringae, controls expression of genes for alginate biosynthesis and is active while the bacteria are associa...

Epidemiology and control of east coast fever in Zambia. A field trial with traditionally managed Sanga cattle.

PubMed

Minjauw, B; Otte, M J; James, A D

1998-06-29

The main objective of the reported field trial was to compare different East Coast Fever (ECF) control strategies for their efficacy, effect on cattle productivity and cost-effectiveness. Five strategies were tested in groups of traditionally managed Sanga cattle over a period of 2.5 years. Two groups were under intensive tick control, one group immunized by the infection and treatment method and the other non-immunized. Two groups were under no tick control, one group immunized and the other non-immunized (the control group). The fifth group was under strategic tick control and was immunized against ECF. All ECF control methods tested significantly reduced mortality, but no marked differences to the control group were seen in other production parameters. No difference in mortality was observed between animals protected from ECF by immunization or by tick control. The most cost-effective method of controlling the disease was by immunization. A financial analysis showed that under the prevailing conditions the break-even price for immunization ranged from US$21.5 to US$25.7 depending of the proportion of reactors. The carrier state induced by immunization did not lead to a persistent high incidence of ECF in non-immunized animals using the same grazing area.

Perception of cattle farmers of the efficacy of east coast fever immunization in southern Zambia.

PubMed

Fandamu, P; Thys, E; Duchateau, L; Berkvens, D

2006-01-01

A study using a structured questionnaire was conducted to assess the perception of cattle farmers of the efficacy of East Coast fever (ECF) immunization in southern Zambia. One hundred and seventy-nine farmers from five districts in southern Zambia were interviewed. The majority of farmers (85%) perceived ECF immunization as being very effective and about half of them (51.4%) preferred immunization to other ECF control strategies. The study showed that the number of calves immunized was strongly associated with the farmer's perception of the benefits of immunization. There was no association between the number of calves immunized and the number of veterinary assistants in a given district or their transport situation. Overall mortality in ECF-immunized calves from various causes stood at 4.2%. Based on farmers' reports, the majority of these deaths seemed to have been caused by anaplasmosis, another tick-borne disease, and might have resulted from relaxation of tick control after ECF immunization. The reasons identified by farmers for not immunizing their animals included failure by immunizing teams to reach certain areas, not having calves of immunization age, and lack of money. These findings provide valuable information on how livestock farmers perceive and adopt new animal disease control strategies and the information could be useful in their planning and implementation.

Percutaneous embolization of persistent low-output enterocutaneous fistulas.

PubMed

Cambj Sapunar, Liana; Sekovski, Budimir; Matić, Dario; Tripković, Andro; Grandić, Leo; Družijanić, Nikica

2012-09-01

To present and retrospectively evaluate the technique of percutaneous embolization of chronic enterocutaneous fistulas (ECFs) using n-butyl-2-cyanoacrylate and Lipiodol under fluoroscopic guidance. Six patients with a total of seven post-operative low-output ECFs of the large intestine were treated. After fistulography a hydrophilic guide wire and a catheter were advanced through the ECF into the intestine. After dilation of the bowel with saline and contrast medium, the catheter was withdrawn into the enteric orifice and glue together with Lipiodol was injected while simultaneously pulling the catheter. Complete closure of all seven fistulas was achieved. There were no peri-procedural complications. In one patient 1 month following embolization a low-output enteric discharge was observed, but the ECF spontaneously healed 5 days later. In one patient 18 months after the embolization a new perforation due to diverticulitis close to the embolization site occurred and resection of the sigmoid colon was performed. One patient needed reoperation due to a recurrence of rectal carcinoma. In our series of patients, the presented technique of percutaneous embolization proved to be efficacious and easy to perform. It may have potential as a first-line treatment of low-output ECFs but a prospective study with a larger series of patients and a longer follow-up is required.

Striking differences in glucose and lactate levels between brain extracellular fluid and plasma in conscious human subjects: effects of hyperglycemia and hypoglycemia.

PubMed

Abi-Saab, Walid M; Maggs, David G; Jones, Tim; Jacob, Ralph; Srihari, Vinod; Thompson, James; Kerr, David; Leone, Paola; Krystal, John H; Spencer, Dennis D; During, Matthew J; Sherwin, Robert S

2002-03-01

Brain levels of glucose and lactate in the extracellular fluid (ECF), which reflects the environment to which neurons are exposed, have never been studied in humans under conditions of varying glycemia. The authors used intracerebral microdialysis in conscious human subjects undergoing electrophysiologic evaluation for medically intractable epilepsy and measured ECF levels of glucose and lactate under basal conditions and during a hyperglycemia-hypoglycemia clamp study. Only measurements from nonepileptogenic areas were included. Under basal conditions, the authors found the metabolic milieu in the brain to be strikingly different from that in the circulation. In contrast to plasma, lactate levels in brain ECF were threefold higher than glucose. Results from complementary studies in rats were consistent with the human data. During the hyperglycemia-hypoglycemia clamp study the relationship between plasma and brain ECF levels of glucose remained similar, but changes in brain ECF glucose lagged approximately 30 minutes behind changes in plasma. The data demonstrate that the brain is exposed to substantially lower levels of glucose and higher levels of lactate than those in plasma; moreover, the brain appears to be a site of significant anaerobic glycolysis, raising the possibility that glucose-derived lactate is an important fuel for the brain.

76 FR 71909 - Television Broadcasting Services; Montgomery, AL

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76 FR 20248 - Television Broadcasting Services; Decatur, IL

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The Role of Extracellular Fluid in Biokinetic Modeling

DOE PAGES

Miller, Guthrie; Klumpp, John A.; Melo, Dunstana; ...

2017-12-01

Here, the pharmacokinetic equations of Pierson et al. describing the behavior of bromide in rat provide a general approach to the modeling of extracellular fluid (ECF). The movement of material into ECF spaces is rapid and is completely characterized by tissue volumes and vascular flow rates to and from a tissue, the volumes of the tissue, and the ECF associated with the tissue. Early-time measurements are needed to characterize ECF. Measurements of DTPA disappearance from plasma by Wedeking et al. are discussed as an example of such measurements. In any biokinetic model, the fastest transfer rates are not determinable withmore » the usual datasets, and if determined empirically, these rates will have very large and highly correlated uncertainties, so particular values of these rates, even though the model fits the available data, are not significant. A pharmacokinetic front-end provides values for these fast rates. An example of such a front-end for a 200–g rat is given.« less

East Coast fever and multiple El Niño Southern oscillation ranks.

PubMed

Fandamu, P; Duchateau, L; Speybroeck, N; Mulumba, M; Berkvens, D

2006-01-30

East Coast fever (ECF), a tick-borne disease of cattle, is a major constraint to livestock development in Africa in general and southern Zambia in particular. Understanding the transmission patterns of this disease complex is very difficult as shown by previous studies in southern and eastern Zambia due to the interplay of risk factors. In this long-term study, we investigated whether global weather changes had any influence on disease transmission in traditionally kept cattle in southern Zambia. The results from this study show a strong association between increased Theileria parva contacts in cattle and the presence of El Niño, clearly linking a simple climatic index to disease outbreaks. We therefore propose that in southern Zambia, the simple and readily available multiple El Niño Southern oscillation index (MEI) ranks be used in planning ECF control programmes and early warning.

78 FR 58470 - Television Broadcasting Services; Cedar Rapids, Iowa

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Is Executive Cognitive Function Associated with Youth Gambling?

PubMed Central

Brodsky, Nancy L.; Brown, Caitlin A.; McKenna, Kathleen A.; Giannetta, Joan M.; Yang, Wei; Romer, Daniel; Hurt, Hallam

2011-01-01

Our objectives for this report were to identify trajectories of youth gambling behavior, and to examine their relation to executive cognitive function (ECF) and associated problem behaviors. Philadelphia school children, enrolled at ages 10–12 years (n = 387; 49% male), completed three annual assessments of risk behaviors, ECF, impulsivity, problem behaviors and demographics. Across ages 10–15 years, using methods from Nagin et al., two groups were identified: Early Gamblers (n = 111) initiated early and continued in later assessments, and Later Gamblers (n = 276) initiated at later ages and gambled less. Betting money on cards and sports were the most frequently reported gambling behaviors. Using gambling group as outcome, final backward selection logistic regression model showed Early Gamblers are more likely male (P = 0.001), report more active coping (P = 0.042), impulsive behaviors (P ≤ 0.008), and have friends who gamble (P = 0.001). Groups were similar in ECF, parental monitoring, marital status, SES, and race. Early Gamblers had higher incidence of problem behaviors and drug use (all P ≤ 0.006). Two gambling groups were identified in early adolescence with Early Gamblers showing higher levels of impulsivity and comorbid problems but similar levels of ECF compared to Late Gamblers. As more gambling groups are identified through later adolescence, ECF may emerge as a relevant precursor of problem gambling at this later time. PMID:21698342

Assessing the impact of East Coast Fever immunisation by the infection and treatment method in Tanzanian pastoralist systems.

PubMed

Martins, S Babo; Di Giulio, G; Lynen, G; Peters, A; Rushton, J

2010-12-01

A field trial was carried out in a Maasai homestead to assess the impact of East Coast Fever (ECF) immunisation by the infection and treatment method (ITM) with the Muguga Cocktail on the occurrence of this disease in Tanzanian pastoralist systems. These data were further used in partial budgeting and decision analysis to evaluate and compare the value of the control strategy. Overall, ITM was shown to be a cost-effective control option. While one ECF case was registered in the immunised group, 24 cases occurred amongst non-immunised calves. A significant negative association between immunisation and ECF cases occurrence was observed (p≤0.001). ECF mortality rate was also lower in the immunised group. However, as anti-theilerial treatment was given to all diseased calves, no significant negative association between immunisation and ECF mortality was found. Both groups showed an overall similar immunological pattern with high and increasing percentages of seropositive calves throughout the study. This, combined with the temporal distribution of cases in the non-immunised group, suggested the establishment of endemic stability. Furthermore, the economic analysis showed that ITM generated a profit estimated to be 7250 TZS (1 USD=1300 TZS) per vaccinated calf, and demonstrated that it was a better control measure than natural infection and subsequent treatment. Copyright © 2010 Elsevier B.V. All rights reserved.

Constraints to cattle production in a semiarid pastoral system in Kenya.

PubMed

Onono, Joshua Orungo; Wieland, Barbara; Rushton, Jonathan

2013-08-01

Livestock keeping is the mainstay for the pastoral community while also providing social and cultural value. This study ranked main production constraints and cattle diseases that impacted livelihood and estimated herd prevalence, incidence rate, and impact of diseases on production parameters in a semiarid pastoral district of Narok in Kenya. Data collection employed participatory techniques including listing, pairwise ranking, disease incidence scoring, proportional piling, and disease impact matrix scoring and this was disaggregated by gender. Production constraints with high scores for impact on livelihood included scarcity of water (19%), lack of extension services (15%), presence of diseases (12%), lack of market for cattle and their products (10%), and recurrent cycle of drought (9%). Diseases with high scores for impact on livelihood were East Coast fever (ECF) (22%) and foot and mouth disease (FMD) (21%). High estimated incidence rates were reported for FMD (67%), trypanosomosis (28%), and ECF (15%), while contagious bovine pleuropneumonia (CBPP) had an incidence rate <1%. Milk yield was affected by FMD, ECF, and trypanosomosis, while ECF was the cause of increased mortality. FMD, ECF, CBPP, and brucellosis caused increased abortion, while effect of gender and location of study was not significant. Despite CBPP being regarded as an important disease affecting cattle production in sub-Sahara Africa, its estimated incidence rate in herds was low. This study indicates what issues should be prioritized by livestock policy for pastoral areas.

Is executive cognitive function associated with youth gambling?

PubMed

Betancourt, Laura M; Brodsky, Nancy L; Brown, Caitlin A; McKenna, Kathleen A; Giannetta, Joan M; Yang, Wei; Romer, Daniel; Hurt, Hallam

2012-06-01

Our objectives for this report were to identify trajectories of youth gambling behavior, and to examine their relation to executive cognitive function (ECF) and associated problem behaviors. Philadelphia school children, enrolled at ages 10-12 years (n = 387; 49% male), completed three annual assessments of risk behaviors, ECF, impulsivity, problem behaviors and demographics. Across ages 10-15 years, using methods from Nagin et al., two groups were identified: Early Gamblers (n = 111) initiated early and continued in later assessments, and Later Gamblers (n = 276) initiated at later ages and gambled less. Betting money on cards and sports were the most frequently reported gambling behaviors. Using gambling group as outcome, final backward selection logistic regression model showed Early Gamblers are more likely male (P = 0.001), report more active coping (P = 0.042), impulsive behaviors (P ≤ 0.008), and have friends who gamble (P = 0.001). Groups were similar in ECF, parental monitoring, marital status, SES, and race. Early Gamblers had higher incidence of problem behaviors and drug use (all P ≤ 0.006). Two gambling groups were identified in early adolescence with Early Gamblers showing higher levels of impulsivity and comorbid problems but similar levels of ECF compared to Late Gamblers. As more gambling groups are identified through later adolescence, ECF may emerge as a relevant precursor of problem gambling at this later time.

[Diagnostic and therapeutic guidelines for entero-cutaneous fistulas. Personal experience and literature review].

PubMed

Candela, G; Di Libero, L; Varriale, S; Manetta, F; Giordano, M; Lanza, M; Argenziano, G; Pizza, A; Sciascia, V; Napolitano, S; Riccio, M; Esposito, D; Santini, L

2007-08-01

The entero-cutaneous fistulas (ECF) are abnormal communications between intestine and abdominal skin. They can occur spontaneously, or after an injury or a surgical procedure. They are associated with a high rate of morbidity and mortality. Spontaneous fistulas can mainly occur in patients affected by cancer, inflammatory bowel disease, diverticulitis, appendicitis, as a result of radiotherapy or injuries. Surgical procedures, carried out in case of neoplastic diseases, inflammatory bowel disease, adhesions removal, represent the primary cause in the development of a postoperative fistulas. Malnourishment, poor general conditions of the patient, high output fistula along with anatomical site of development, and the presence of abscesses, represent the negative factors influencing the spontaneous healing of fistulas. The experience reported here is about three ECF cases occurred after surgery and treated only with medical therapy. The first case is a woman in good general conditions who underwent surgery to remove a recurrent retroperitoneal myxoid liposarcoma situated in the right lower quadrant. The patient had never undergone surgery for an intestinal resection. The other two patients analyzed were affected by sepsis and metabolic unbalance and had developed a fistula after colonic resection. Fluids and electrolytes adjustments and sepsis management have preceded any other kind of therapy. Continuous infusion with somatostatin, fast, proton pump inhibitors and loperamide have been taken up to decrease secretions and intestinal motility. Total parenteral nutrition has been essential to recover nutritional status and improve patients' general conditions. In order to heal and protect peri-fistula skin we have used sterile washing solutions, absorbable ionic exchange resin, silver and polyurethanes based medications and colostomy bags adhesive systems. Since surgical treatment of ECF is associated with high rates of morbidity and mortality, conservative treatment should always be taken into consideration. When conservative treatment fails, delayed surgical intervention has been related to a higher rate of success. The purpose of this study is to describe diagnostic and therapeutic guidelines to general surgeons, like ourselves, whenever they have to deal with ECF cases.

Climate change and cattle vector-borne diseases: Use of participatory epidemiology to investigate experiences in pastoral communities in Northern Tanzania.

PubMed

Kimaro, Esther G; Toribio, Jenny-Ann L M L; Mor, Siobhan M

2017-11-01

Climate change is predicted to increase incidence of vector-borne diseases in humans, however, little is known about the impact of such diseases in livestock. In the absence of historical data with which to examine the inter-relation between climate and disease, participatory epidemiological (PE) methods were used with Maasai pastoralists of Monduli District, northern Tanzania to establish local observations on two major vector-borne diseases of cattle, namely East Coast fever (ECF) and African animal trypanosomiasis (AAT). Data collection involving gender segregated groups (10 men groups and 9 women groups) occurred in 10 randomly selected villages between November 2014 and March 2015. ECF and AAT were ranked amongst the top 5 most important cattle diseases with strong agreement across informant groups (Kendall's W=0.40 for men and 0.45 for women; p<0.01). Matrix scoring for both men and women groups confirmed that Masaai easily recognize these diseases. All groups associated ECF with the wet and cool dry seasons. AAT was more variable throughout the year, with more cases reported in the long dry season. Likewise, pastoralists reported differences in seasonal occurrence of disease vectors (Rhipicephalus appendiculatus and Glossina spp.) by village. Comparing 2014-1984, participant groups consistently reported declines in rainfall, vegetation cover and quality pasture, as well as increases in severe droughts. Experiences with ECF/AAT and vector abundance between these time periods was more variable across villages, and likely relates to changes in climate and animal management practices over the last 30 years. This baseline study is the first to document the inter-relation between climate and cattle vector-borne disease from the pastoralist perspective. Findings from this study reveal a complex interplay between human, animal and environmental factors, understanding of which is urgently required to devise approaches to mitigate effects of climate change in these vulnerable areas. Copyright © 2017 Elsevier B.V. All rights reserved.

Identification of Breast Cancer Specific Proteolytic Activities for Targeted Prodrug Activation

DTIC Science & Technology

2006-05-01

volume of fluid that can be obtained from ECF of human breast cancers is to use a phage display approach. To accomplish this, we have designed a...affinity support, followed by a randomized protease substrate sequence and the carboxyl-terminal domain of M13 gene III. Each fusion protein was displayed ...PSMA) (35). Substrate phage can be created either as a monovalent or as pentavalent display (34). Both approaches have their own advantages and

Identification of the CvsSR regulon in Pseudomonas syringae reveals overlap with the Type-III secretion and AlgU regulons

USDA-ARS?s Scientific Manuscript database

Pseudomonas syringae pv. tomato DC3000 (Pto) lives epiphytically and endophytically during its infection cycle. Two-component systems (TCSs) and extracytoplasmic function (ECF) sigma factors are used by Pto to sense environmental changes within the leaf apoplast during pathogenesis. The TCS, CvsSR i...

Case Study of Tenure-Track Early Career Faculty in a College of Education

ERIC Educational Resources Information Center

Esping, Gretchen Revay

2010-01-01

This dissertation examines an understudied group according to the American Council on Education: the tenure-track early career faculty (ECF). The focus is on the culturalization, socialization, academic culture, and emergent themes discerned from ten semi-structured interviews with tenure-track ECF. This qualitative bounded system case study…

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Absolute quantification of the host-to-parasite DNA ratio in theileria parva-infected lymphocyte cell lines

USDA-ARS?s Scientific Manuscript database

Theileria parva is a tick-transmitted intracellular apicomplexan pathogen of cattle in sub-Saharan Africa that causes East Coast fever (ECF). ECF is an acute fatal disease that kills over one million cattle annually, imposing a tremendous burden on African small-holder cattle farmers. The pathology ...

Immunization of cattle against East Coast fever using Theileria parva (Marikebuni) and relaxation of tick control in North Rift, Kenya.

PubMed

Wanjohi, J M; Ngeranwa, J N; Rumberia, R M; Muraguri, G R; Mbogo, S K

2001-09-01

A total of 90 animals was immunized against East Coast fever (ECF) using Theileria parva (Marikebuni) stock on three large-scale farms in Kiminini Division, Trans-Nzoia District, North Rift, Kenya. Another 90 cattle served as non-immunized controls. Following immunization the number of cattle with significant indirect fluorescent antibody (IFA) titres increased from 43.9% to 84.4% and 6.7% of the cattle developed clinical ECF reactions. Two months after immunization, the immunized and non-immunized cattle were divided into two groups one of which was dipped every 3 weeks and the other dipped when total full body tick counts reached 100. All the animals were monitored for 51 weeks for incidences of ECF and other tick-borne diseases. Twenty-four cases of ECF were diagnosed among the non-immunized cattle compared to four cases among the immunized cattle; a difference that was significant (P > 0.05). There was no significant difference in the incidences of babesiosis and anaplasmosis between the immunized and non-immunized cattle.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Spriggs, G D

In a previous paper, the composite exposure rate conversion factor (ECF) for nuclear fallout was calculated using a simple theoretical photon-transport model. The theoretical model was used to fill in the gaps in the FGR-12 table generated by ORNL. The FGR-12 table contains the individual conversion factors for approximate 1000 radionuclides. However, in order to calculate the exposure rate during the first 30 minutes following a nuclear detonation, the conversion factors for approximately 2000 radionuclides are needed. From a human-effects standpoint, it is also necessary to have the dose rate conversion factors (DCFs) for all 2000 radionuclides. The DCFs aremore » used to predict the whole-body dose rates that would occur if a human were standing in a radiation field of known exposure rate. As calculated by ORNL, the whole-body dose rate (rem/hr) is approximately 70% of the exposure rate (R/hr) at one meter above the surface. Hence, the individual DCFs could be estimated by multiplying the individual ECFs by 0.7. Although this is a handy rule-of-thumb, a more consistent (and perhaps, more accurate) method of estimating the individual DCFs for the missing radionuclides in the FGR-12 table is to use the linear relationship between DCF and total gamma energy released per decay. This relationship is shown in Figure 1. The DCFs for individual organs in the body can also be estimated from the estimated whole-body DCF. Using the DCFs given FGR-12, the ratio of the organ-specific DCFs to the whole-body DCF were plotted as a function of the whole-body DCF. From these plots, the asymptotic ratios were obtained (see Table 1). Using these asymptotic ratios, the organ-specific DCFs can be estimated using the estimated whole-body DCF for each of the missing radionuclides in the FGR-12 table. Although this procedure for estimating the organ-specific DCFs may over-estimate the value for some low gamma-energy emitters, having a finite value for the organ-specific DCFs in the table is probably better than having no value at all. A summary of the complete ECF and DCF values are given in Table 2.« less

Alcohol affects executive cognitive functioning differentially on the ascending versus descending limb of the blood alcohol concentration curve.

PubMed

Pihl, Robert O; Paylan, Sheila S; Gentes-Hawn, Alyson; Hoaken, Peter N S

2003-05-01

Executive cognitive functioning (ECF), a construct that includes cognitive abilities such as planning, abstract reasoning, and the capacity to govern self-directed behavior, has been recently researched as an antecedent to many forms of psychopathology and has been implicated in alcohol-related aggression. This study was designed to examine whether differential ECF impairments can be noted on the ascending versus the descending limbs of the blood alcohol concentration curve. Forty-one male university students participated in this study. Twenty-one subjects were given 1.32 ml of 95% alcohol per kilogram of body weight, mixed with orange juice, and the remaining 20 were given a placebo. Participants were randomly assigned to either an ascending or descending blood alcohol group and were tested on six tests of ECF on their assigned limb. Subjective mood data were also collected. Intoxicated participants on both limbs demonstrated ECF impairment; the descending-limb group showed greater impairment than their ascending-limb counterparts. Intoxicated subjects were significantly more anxious at baseline than placebo subjects. The introduction of this covariate nullified any significant differences in subjective mood found on either limb of the blood alcohol concentration curve, but ECF impairments remained robust. Our results support the conclusion that alcohol negatively affects cognitive performance and has a differential effect on the descending versus the ascending limb of the blood alcohol concentration curve. The latter finding may have important ramifications relating to the detrimental consequences of alcohol intoxication.

Elicitation of gymnemic acid production in cell suspension cultures of Gymnema sylvestre R.Br. through endophytic fungi.

PubMed

Netala, Vasudeva Reddy; Kotakadi, Venkata Subbaiah; Gaddam, Susmila Aparna; Ghosh, Sukhendu Bikash; Tartte, Vijaya

2016-12-01

The enhancement of plant secondary metabolite production in cell suspension cultures through biotic or abiotic elicitation has become a potential biotechnological approach for commercialization or large-scale production of bioactive compounds. Gymnema sylvestre R.Br. is an important medicinal plant, rich in a group of oleanane triterpenoid saponins called gymnemic acid, well known for its anti-diabetic activity. Two endophytic fungal strains were isolated from the leaves of G. sylvestre and identified as Polyancora globosa and Xylaria sp. based on the PCR amplification and internal transcribed spacer (ITS 1-5.8S-ITS 2) sequencing of 18S rRNA gene. The process of elicitation of cell suspension cultures of G. sylvestre with dried powder of fungal mycelia (DPFM) and extracellular culture filtrate (ECF) of endophytic fungi consistently enhanced the accumulation of gymnemic acid and the DPFM was proved to be an effective elicitor when compared to the ECF. The DPFM elicited the gymnemic acid content in the range of 2.57-10.45-fold, while the ECF elicited the gymnemic acid content in the range of 2.39-7.8-fold. P. globosa, a novel and a rare endophytic fungal strain, has shown a great influence on the production of gymnemic acid. Cell suspension cultures elicited with DPFM of P. globosa produced higher amount of gymnemic acid content (124.23 mg/g dried cell weight) compared to the cultures elicited with DPFM of Xylaria sp. (102.24 mg/g DCW). But the cultures treated with consortium of DPFM of both fungi showed great influence on the production of gymnemic acid (139.98 mg/g DCW) than the cultures treated with DPFM alone. Similarly, cultures treated with consortium of ECF of both fungi produced more gymnemic acid content (94.86 mg/g DCW) compared with cultures treated with ECF of Xylaria sp. (77.93 mg/g DCW) and ECF of P. globosa (78.65 mg/g DCW) alone.

77 FR 9653 - Comment Sought on Potential Data for Connect America Fund Phase One Incremental Support

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2012-02-17

... Internet by accessing the ECFS: http://fjallfoss.fcc.gov/ecfs2/ . Paper Filers: Parties who choose to file..., or via the Internet at http://www.bcpiweb.com . 1. On November 18, 2011, the Commission released the... things, the Commission established a transitional mechanism to distribute high cost universal service...

Successful embolization of a enterocutaneous fistula tract with Onyx 34 following low anterior resection for rectal cancer.

PubMed

Rahimi, Hamza; Venbrux, Anthony C; Obias, Vincent

2018-06-01

Enterocutaneous fistulas (ECFs) can be one of the complications found after surgical intervention for rectal cancer. Interventional modalities consisting of surgical, endoscopic, and radiological methods are often implemented to treat postoperative symptomatic complications. We present the case of 61-year-old Caucasian man who presented to us with a recent diagnosis of rectal cancer that had invaded the levators as well as anteriorly into the prostate, and who underwent low anterior resection with a diverting loop ileostomy. The patient was found to have a persistent presacral abscess due to an ECF tract. This case highlights the off-label use of ethylene-vinyl alcohol copolymer dissolved in dimethyl sulfoxide (Onyx 34) to seal an ECF.

Removal of natural organic matter and arsenic from water by electrocoagulation/flotation continuous flow reactor.

PubMed

Mohora, Emilijan; Rončević, Srdjan; Dalmacija, Božo; Agbaba, Jasmina; Watson, Malcolm; Karlović, Elvira; Dalmacija, Milena

2012-10-15

The performance of the laboratory scale electrocoagulation/flotation (ECF) reactor in removing high concentrations of natural organic matter (NOM) and arsenic from groundwater was analyzed in this study. An ECF reactor with bipolar plate aluminum electrodes was operated in the horizontal continuous flow mode. Electrochemical and flow variables were optimized to examine ECF reactor contaminants removal efficiency. The optimum conditions for the process were identified as groundwater initial pH 5, flow rate=4.3 l/h, inter electrode distance=2.8 cm, current density=5.78 mA/cm(2), A/V ratio=0.248 cm(-1). The NOM removal according to UV(254) absorbance and dissolved organic matter (DOC) reached highest values of 77% and 71% respectively, relative to the raw groundwater. Arsenic removal was 85% (6.2 μg As/l) relative to raw groundwater, satisfying the drinking water standards. The specific reactor electrical energy consumption was 17.5 kWh/kg Al. The specific aluminum electrode consumption was 66 g Al/m(3). According to the obtained results, ECF in horizontal continuous flow mode is an energy efficient process to remove NOM and arsenic from groundwater. Copyright © 2012 Elsevier B.V. All rights reserved.

The sigmaR regulon of Streptomyces coelicolor A32 reveals a key role in protein quality control during disulphide stress.

PubMed

Kallifidas, Dimitris; Thomas, Derek; Doughty, Phillip; Paget, Mark S B

2010-06-01

Diamide is an artificial disulphide-generating electrophile that mimics an oxidative shift in the cellular thiol-disulphide redox state (disulphide stress). The Gram-positive bacterium Streptomyces coelicolor senses and responds to disulphide stress through the sigma(R)-RsrA system, which comprises an extracytoplasmic function (ECF) sigma factor and a redox-active anti-sigma factor. Known targets that aid in the protection and recovery from disulphide stress include the thioredoxin system and genes involved in producing the major thiol buffer mycothiol. Here we determine the global response to diamide in wild-type and sigR mutant backgrounds to understand the role of sigma(R) in this response and to reveal additional regulatory pathways that allow cells to cope with disulphide stress. In addition to thiol oxidation, diamide was found to cause protein misfolding and aggregation, which elicited the induction of the HspR heat-shock regulon. Although this response is sigma(R)-independent, sigma(R) does directly control Clp and Lon ATP-dependent AAA(+) proteases, which may partly explain the reduced ability of a sigR mutant to resolubilize protein aggregates. sigma(R) also controls msrA and msrB methionine sulphoxide reductase genes, implying that sigma(R)-RsrA is responsible for the maintenance of both cysteine and methionine residues during oxidative stress. This work shows that the sigma(R)-RsrA system plays a more significant role in protein quality control than previously realized, and emphasizes the importance of controlling the cellular thiol-disulphide redox balance.

Cystic fibrosis: Beyond the airways. Report on the meeting of the basic science working group in Loutraki, Greece.

PubMed

Amaral, Margarida D; Boj, Sylvia F; Shaw, James; Leipziger, Jens; Beekman, Jeffrey M

2018-06-01

The European Cystic Fibrosis Society (ECFS) Basic Science Working Group (BSWG) organized a session on the topic "Cystic Fibrosis: Beyond the Airways", within the 15th ECFS Basic Science Conference which gathered around 200 researchers working in the basic science of CF. The session was organized and chaired by Margarida Amaral (BioISI, University of Lisboa, Portugal) and Jeffrey Beekman (University Medical Centre Utrecht, Netherlands) as Chair and Vice-Chair of the BSWG and its purpose was to bring attention of participants of the ECFS Basic Science Conference to "more forgotten" organs in CF disease. In this report we attempt to review and integrate the ideas that emerged at the session. Copyright © 2018 European Cystic Fibrosis Society. All rights reserved.

Characteristics of Resting-State Functional Connectivity in Intractable Unilateral Temporal Lobe Epilepsy Patients with Impaired Executive Control Function

PubMed Central

Zhang, Chao; Yang, Hongyu; Qin, Wen; Liu, Chang; Qi, Zhigang; Chen, Nan; Li, Kuncheng

2017-01-01

Executive control function (ECF) deficit is a common complication of temporal lobe epilepsy (TLE). Characteristics of brain network connectivity in TLE with ECF dysfunction are still unknown. The aim of this study was to investigate resting-state functional connectivity (FC) changes in patients with unilateral intractable TLE with impaired ECF. Forty right-handed patients with left TLE confirmed by comprehensive preoperative evaluation and postoperative pathological findings were enrolled. The patients were divided into normal ECF (G1) and decreased ECF (G2) groups according to whether they showed ECF impairment on the Wisconsin Card Sorting Test (WCST). Twenty-three healthy volunteers were recruited as the healthy control (HC) group. All subjects underwent resting-state functional magnetic resonance imaging (rs-fMRI). Group-information-guided independent component analysis (GIG-ICA) was performed to estimate resting-state networks (RSNs) for all subjects. General linear model (GLM) was employed to analyze intra-network FC (p < 0.05, false discovery rate, FDR correction) and inter-network FC (p < 0.05, Bonferroni correction) of RSN among three groups. Pearson correlations between FC and neuropsychological tests were also determined through partial correlation analysis (p < 0.05). Eleven meaningful RSNs were identified from 40 left TLE and 23 HC subjects. Comparison of intra-network FC of all 11 meaningful RSNs did not reveal significant difference among the three groups (p > 0.05, FDR correction). For inter-network analysis, G2 exhibited decreased FC between the executive control network (ECN) and default-mode network (DMN) when compared with G1 (p = 0.000, Bonferroni correction) and HC (p = 0.000, Bonferroni correction). G1 showed no significant difference of FC between ECN and DMN when compared with HC. Furthermore, FC between ECN and DMN had significant negative correlation with perseverative responses (RP), response errors (RE) and perseverative errors (RPE) and had significant positive correlation categories completed (CC) in both G1 and G2 (p < 0.05). No significant difference of Montreal Cognitive Assessment (MoCA) was found between G1 and G2, while intelligence quotient (IQ) testing showed significant difference between G1and G2.There was no correlation between FC and either MoCA or IQ performance. Our findings suggest that ECF impairment in unilateral TLE is not confined to the diseased temporal lobe. Decreased FC between DMN and ECN may be an important characteristic of RSN in intractable unilateral TLE. PMID:29375338

75 FR 63052 - Airworthiness Directives; Eurocopter France (ECF) Model AS350B3 and EC130 B4 Helicopters

Federal Register 2010, 2011, 2012, 2013, 2014

2010-10-14

... Airworthiness Directives; Eurocopter France (ECF) Model AS350B3 and EC130 B4 Helicopters AGENCY: Federal... directive (AD) for the Eurocopter France Model AS350B3 and EC130 B4 helicopters. This amendment is prompted... autorotation training making it impossible to execute a power recovery and compelling the pilot to continue the...

Quantification of extracellular matrix expansion by CMR in infiltrative heart disease.

PubMed

Mongeon, François-Pierre; Jerosch-Herold, Michael; Coelho-Filho, Otávio Rizzi; Blankstein, Ron; Falk, Rodney H; Kwong, Raymond Y

2012-09-01

The aim of this study was to perform direct quantification of myocardial extracellular volume fraction (ECF) with T1-weighted cardiac magnetic resonance (CMR) imaging in patients suspected to have infiltrative heart disease. Infiltrative heart disease refers to accumulation of abnormal substances within the myocardium. Qualitative assessment of late gadolinium enhancement (LGE) remains the most commonly used method for CMR evaluation of patients suspected with myocardial infiltration. This technique is widely available and can be performed in a reproducible and standardized manner. However, the degree of extracellular matrix expansion due to myocardial infiltration in the intercellular space has, to date, not been amenable to noninvasive quantification with LGE. We performed 3-T CMR in 38 patients (mean age 68 ± 15 years) who were referred for assessment of infiltrative heart disease and also in 9 healthy volunteers as control subjects. The T1 quantification by Look-Locker gradient-echo before and after contrast determined segmental myocardial partition coefficients. The ECF was obtained by referencing the tissue partition coefficient for gadolinium to the plasma volume fraction in blood, derived from serum hematocrit. Cine CMR and LGE imaging in matching locations were also performed. Seventeen patients (45%) had cardiac amyloidosis (CA) (biopsy-confirmed or clinically highly probable), 20 (53%) had a non-amyloid cardiomyopathy, and 1 had lysosomal storage disease. Median global ECF was substantially higher in CA patients (0.49) compared with non-amyloid cardiomyopathy patients (0.33, p < 0.0001) and volunteers (0.24, p = 0.0001). The ECF strongly correlated with visually assessed segmental LGE (r = 0.80, p < 0.0001) and LV mass index (r = 0.69, p < 0.0001), reflecting severity of myocardial infiltration. In patients with CA, ECF was highest in segments with LGE, although it remained elevated in segments without qualitative LGE. The CMR ECF quantification identified substantial expansion of the interstitial space in patients with CA compared with volunteers. Further studies using this technique for diagnosis and assessment of the severity of myocardial infiltration are warranted. Copyright © 2012 American College of Cardiology Foundation. Published by Elsevier Inc. All rights reserved.

Improvement in rice straw pulp bleaching effluent quality by incorporating oxygen delignification stage prior to elemental chlorine-free bleaching.

PubMed

Kaur, Daljeet; Bhardwaj, Nishi K; Lohchab, Rajesh Kumar

2017-10-01

Environmental degradation by industrial and other developmental activities is alarming for imperative environmental management by process advancements of production. Pulp and paper mills are now focusing on using nonwood-based raw materials to protect forest resources. In present study, rice straw was utilized for pulp production as it is easily and abundantly available as well as rich in carbohydrates (cellulose and hemicelluloses). Soda-anthraquinone method was used for pulp production as it is widely accepted for agro residues. Bleaching process during paper production is the chief source of wastewater generation. The chlorophenolic compounds generated during bleaching are highly toxic, mutagenic, and bioaccumulative in nature. The objectives of study were to use oxygen delignification (ODL) stage prior to elemental chlorine-free (ECF) bleaching to reduce wastewater load and to study its impact on bleached pulp characteristics. ODL stage prior to ECF bleaching improved the optical properties of pulp in comparison to only ECF bleaching. When ODL stage was incorporated prior to bleaching, the tensile index and folding endurance of the pulp were found to be 56.6 ± 1.5 Nm/g and 140, respectively, very high in comparison to ECF alone. A potential reduction of 51, 57, 43, and 53% in BOD 3 , COD, color, and AOX, respectively was observed on adding the ODL stage compared to ECF only. Generation of chlorophenolic compounds was reduced significantly. Incorporation of ODL stage prior to bleaching was found to be highly promising for reducing the toxicity of bleaching effluents and may lead to better management of nearby water resources. Graphical abstract ᅟ.

The determinants of transverse tubular volume in resting skeletal muscle

PubMed Central

Sim, Jingwei; Fraser, James A

2014-01-01

The transverse tubular (t)-system of skeletal muscle couples sarcolemmal electrical excitation with contraction deep within the fibre. Exercise, pathology and the composition of the extracellular fluid (ECF) can alter t-system volume (t-volume). T-volume changes are thought to contribute to fatigue, rhabdomyolysis and disruption of excitation–contraction coupling. However, mechanisms that underlie t-volume changes are poorly understood. A multicompartment, history-independent computer model of rat skeletal muscle was developed to define the minimum conditions for t-volume stability. It was found that the t-system tends to swell due to net ionic fluxes from the ECF across the access resistance. However, a stable t-volume is possible when this is offset by a net efflux from the t-system to the cell and thence to the ECF, forming a net ion cycle ECF→t-system→sarcoplasm→ECF that ultimately depends on Na+/K+-ATPase activity. Membrane properties that maximize this circuit flux decrease t-volume, including PNa(t) > PNa(s), PK(t) < PK(s) and N(t) < N(s) [P, permeability; N, Na+/K+-ATPase density; (t), t-system membrane; (s), sarcolemma]. Hydrostatic pressures, fixed charges and/or osmoles in the t-system can influence the magnitude of t-volume changes that result from alterations in this circuit flux. Using a parameter set derived from literature values where possible, this novel theory of t-volume was tested against data from previous experiments where t-volume was measured during manipulations of ECF composition. Predicted t-volume changes correlated satisfactorily. The present work provides a robust, unifying theoretical framework for understanding the determinants of t-volume. PMID:25384782

Isolating isomers of perfluorocarboxylates in polar bears (Ursus maritimus) from two geographical locations.

PubMed

De Silva, Amila O; Mabury, Scott A

2004-12-15

The source of involatile, anthropogenic perfluorocarboxylate anions (PFCAs) in biota from remote regions is of heightened interest due to the persistence, toxicity, and bioaccumulation of these materials. Large-scale production of fluorinated compounds is carried out primarily by one of two methods: electrochemical fluorination (ECF) and telomerization. Products of the two processes may be distinguished based on constitutional isomer pattern as ECF products are characteristically comprised of a variety of constitutional isomers. The objective of this research was to develop a method for identifying the constitutional isomer profile of PFCAs in environmental samples and to apply the method to polar bear livers from two different locations. Resolution of constitutional isomers of derivatized PFCAs (8-13 carbons) was accomplished via GC-MS. Seven isomers of an authentic ECF perfluorooctanoate (PFOA) standard were separated. The linear isomer comprised 78% of this standard. Isomer profiles of PFCAs in liver samples of 15 polar bears (Ursus maritimus) from the Canadian Arctic and eastern Greenland were determined by GC-MS. The PFOA isomer pattern in Greenland polar bear samples showed a variety of branched isomers while only the linear PFOA isomer was determined in Canadian samples. Samples of both locations had primarily (>99%) linear isomers of perfluorononanoate and perfluorotridecanoate. Branched isomers of perfluorodecanoate, perfluoroundecanoate, and perfluorododecanoate were determined in the polar bear samples. Unlike the PFOA isomer signature, only a single branched isomer peak on the chromatograms was observed for these longer chain PFCAs. The presence of branched isomers suggests some contribution from ECF sources. However, in comparison to the amount of branched isomers in the ECF PFOA standard, such minor percentages of branched PFCAs may suggest additional input from an exclusively linear isomer source.

Increased Urine Production Due to Leg Fluid Displacement Reduces Hours of Undisturbed Sleep.

PubMed

Kiba, Keisuke; Hirayama, Akihide; Yoshikawa, Motokiyo; Yamamoto, Yutaka; Torimoto, Kazumasa; Shimizu, Nobutaka; Tanaka, Nobumichi; Fujimoto, Kiyohide; Uemura, Hirotsugu

2017-07-03

To investigate whether or not the leg fluid displacement observed when moving from the standing to recumbent position at bedtime reduces the hours of undisturbed sleep (HUS). Men aged 50 years or older who were hospitalized for urological diseases were investigated. Body water evaluation was performed three times with a bioelectric impedance method: (i) 17:00, (ii) 30 min after (short-term), and (iii) waking up (long-term). A frequency volume chart was used to evaluate the status of nocturnal urine production, and the factors affecting HUS were investigated. A total of 50 patients (mean age: 68 years) were enrolled. Short-term changes in extracellular fluid (ECF in the legs showed a significant positive correlation with urine production per unit of time at the first nocturnal voiding (UFN/HUS) (r = 0.45, P = 0.01). In the comparison between patients who had <3 HUS vs. those who had ≥3 HUS, the <3 HUS group showed significantly greater short-term changes in leg fluid volume, night-time water intake (17:00-06:00), and UFN/HUS. Multivariate analysis to assess the risk factors for <3 HUS indicated UFN/HUS as a risk factor in the overall model, and short-term changes in leg ECF and night-time water intake as risk factors in the model that only considered factors before sleep. Nocturnal leg fluid displacement may increase urine production leading up to first voiding after going to bed, and consequently, induce early awakening after falling asleep. © 2017 John Wiley & Sons Australia, Ltd.

Advances in the treatment of gastric cancer.

PubMed

Ilson, David H

2017-11-01

To review recent studies in esophagogastric cancer. Positive emission tomography (PET) scan in follow-up after curative treatment of esophagogastric cancer did not lead to improved survival. In the preoperative treatment of esophagogastric cancer, the addition of the antivascular endothelial growth factor agent bevacizumab to perioperative chemotherapy with combination epirubicin, cisplatinum, and 5-fluorouracil (5-FU; ECF) failed to improve survival compared with chemotherapy alone. In a head-to-head comparison of preoperative chemotherapy for locally advanced gastric and esophagogastric adenocarcinoma, FLOT (fluorouracil, leucovorin, oxaliplatin, and docetaxel) significantly improved overall survival compared with ECF. Assessing response to induction chemotherapy prior to combined preoperative chemoradiotherapy in PET nonresponding patients allowed a change in chemotherapy during subsequent radiotherapy with improved rates of pathologic complete response. In human epidermal growth factor receptor-2-positive advanced esophagogastric adenocarcinoma, second-line treatment with the chemotherapy/trastuzumab drug conjugate emtansine/trastuzumab failed to improve response or overall survival compared with treatment using paclitaxel chemotherapy. The immune checkpoint inhibitor, nivolumab, improved survival in refractory gastric cancer. Recent studies in gastric cancer clarify the optimal preoperative chemotherapy regimen and the use of PET scan as a response measure of preoperative therapy in esophagogastric cancer, and the role of targeted agents and immune checkpoint inhibitors in metastatic disease.

Alkaline peroxide treatment of ECF bleached softwood kraft pulps: part 2. effect of increased fiber charge on refining, wet-end application, and hornification

Treesearch

Zheng Dang; Thomas Elder; Jeffery S. Hsieh; Arthur J. Ragauskas

2007-01-01

The effect of increased fiber charge on refining, cationic starch adsorption, and hornification was examined. Two pulps were investigated: (1) a softwood (SW) kraft pulp (KP) which was bleached elementally chlorine-free (ECF) and sewed as control; and (2) a control pulp treated with alkaline peroxide, which had a higher fiber charge. It was shown that increased fiber...

Nutritional management of enterocutaneous fistulas.

PubMed

Slater, Rebecca

Enterocutaneous fistulas (ECF) may be challenging to manage due to the large volume of fluid losses, that may result in severe dehydration, electrolyte imbalances, malnutrition and sepsis. It is imperative that this group of patients receive adequate nutrition, as malnutrition and sepsis are the leading cause of death. ECF treatment is complex and based on various assessments, treatment can be medical/conservative management or surgical. Depending on the site of the fistula and the nutritional status of the patient, clinicians have to decide whether parenteral nutrition or enteral nutrition should be established. Fistuloclysis is a relatively novel procedure in which nutrition is provided via an enteral feeding tube placed directly into the distal lumen of a high output fistula. Although fistuloclysis is not feasible for all patients with ECF, for those that are eligible, the method appears to be an acceptable and safe method of maintaining and improving nutritional status.

Effect of different East Coast Fever control strategies on fertility, milk production and weight gain of Sanga cattle in the Central Province of Zambia.

PubMed

Minjauw, B; Otte, J; James, A D; de Castro, J J; Sinyangwe, P

1997-12-01

Five different East Coast Fever (ECF) (Theileria parva infection) control strategies, based on tick control and/or ECF immunization, were tested in groups of traditionally managed Sanga cattle in the Central Province of Zambia over a period of 2.5 years. Two groups were kept under intensive tick control (sprayed weekly), one group immunized and one non-immunized. Two further groups were under no tick control, one group immunized and one non-immunized, while a fifth group was immunized against ECF and maintained under strategic tick control (18 sprays per year). Tick control increased milk production and weight gain but not fertility. Immunization had neither marked detrimental nor beneficial effects on the cattle productivity. The combination of strategic tick control and immunization resulted in the highest level of production and at the same time reduced the potential risk from other tick-borne diseases.

Theileria parva seroprevalence in traditionally kept cattle in southern Zambia and El Nino.

PubMed

Fandamu, P; Duchateau, L; Speybroeck, N; Marcotty, T; Mbao, V; Mtambo, J; Mulumba, M; Berkvens, D

2005-04-01

Sero-epidemiological surveys involving 27,526 cattle over a period of 8 years show that Theileria parva, the parasite causing East Coast fever (ECF) is found throughout southern Zambia. Higher values of T. parva sero-prevalence were observed in the plateau districts of Monze, Choma and Mazabuka than in the valley districts of Siavonga and Sinazongwe. Our results reveal a strong association between high T. parva sero-prevalence and the presence of the periodic climatic phenomenon known as the El Nino Southern Oscillation. More T. parva sero-positive samples were recorded during El Nino years (1997/98) (P<0.001) than other years in the study period. From this association, we conclude that Multiple El Nino Southern Oscillation Indices can be used to predict years with high or low ECF infection prevalence thereby contributing to the improved control of ECF in the area.

Role of ATP-binding cassette and solute carrier transporters in erlotinib CNS penetration and intracellular accumulation.

PubMed

Elmeliegy, Mohamed A; Carcaboso, Angel M; Tagen, Michael; Bai, Feng; Stewart, Clinton F

2011-01-01

To study the role of drug transporters in central nervous system (CNS) penetration and cellular accumulation of erlotinib and its metabolite, OSI-420. After oral erlotinib administration to wild-type and ATP-binding cassette (ABC) transporter-knockout mice (Mdr1a/b(-/-), Abcg2(-/-), Mdr1a/b(-/-)Abcg2(-/-), and Abcc4(-/-)), plasma was collected and brain extracellular fluid (ECF) was sampled using intracerebral microdialysis. A pharmacokinetic model was fit to erlotinib and OSI-420 concentration-time data, and brain penetration (P(Brain)) was estimated by the ratio of ECF-to-unbound plasma area under concentration-time curves. Intracellular accumulation of erlotinib was assessed in cells overexpressing human ABC transporters or SLC22A solute carriers. P(Brain) in wild-type mice was 0.27 ± 0.11 and 0.07 ± 0.02 (mean ± SD) for erlotinib and OSI-420, respectively. Erlotinib and OSI-420 P(Brain) in Abcg2(-/-) and Mdr1a/b(-/-)Abcg2(-/-) mice were significantly higher than in wild-type mice. Mdr1a/b(-/-) mice showed similar brain ECF penetration as wild-type mice (0.49 ± 0.37 and 0.04 ± 0.02 for erlotinib and OSI-420, respectively). In vitro, erlotinib and OSI-420 accumulation was significantly lower in cells overexpressing breast cancer resistance protein (BCRP) than in control cells. Only OSI-420, not erlotinib, showed lower accumulation in cells overexpressing P-glycoprotein (P-gp) than in control cells. The P-gp/BCRP inhibitor elacridar increased erlotinib and OSI-420 accumulation in BCRP-overexpressing cells. Erlotinib uptake was higher in OAT3- and OCT2-transfected cells than in empty vector control cells. Abcg2 is the main efflux transporter preventing erlotinib and OSI-420 penetration in mouse brain. Erlotinib and OSI-420 are substrates for SLC22A family members OAT3 and OCT2. Our findings provide a mechanistic basis for erlotinib CNS penetration, cellular uptake, and efflux mechanisms. ©2010 AACR.

Lipotoxin F of Pseudomonas aeruginosa is an AlgU-dependent and alginate-independent outer membrane protein involved in resistance to oxidative stress and adhesion to A549 human lung epithelia.

PubMed

Damron, F Heath; Napper, Jennifer; Teter, M Allison; Yu, Hongwei D

2009-04-01

Chronic lung infection with P. aeruginosa and excessive neutrophil-associated inflammation are major causes of morbidity and mortality in patients with cystic fibrosis (CF). Overproduction of an exopolysaccharide known as alginate leads to the formation of mucoid biofilms that are resistant to antibiotics and host defences. Alginate overproduction or mucoidy is controlled by a stress-related ECF sigma factor AlgU/T. Mutation in the anti-sigma factor MucA is a known mechanism for conversion to mucoidy. Recently, we showed that inactivation of a kinase (KinB) in nonmucoid strain PAO1 results in overproduction of alginate. Here, we report the initial characterization of lipotoxin F (LptF, PA3692), an OmpA-like outer membrane protein that exhibited increased expression in the mucoid PAO1kinB mutant. The lipotoxin family of proteins has been previously shown to induce inflammation in lung epithelia, which may play a role in CF disease progression. Expression of LptF was observed to be AlgU-dependent and upregulated in CF isolates. Deletion of lptF from the kinB mutant had no effect on alginate production. Deletion of lptF from PAO1 caused a differential susceptibility to oxidants that can be generated by phagocytes. The lptF and algU mutants were more sensitive to hypochlorite than PAO1. However, the lptF mutant displayed increased resistance to hydrogen peroxide. LptF also contributed to adhesion to A549 human lung epithelial cells. Our data suggest that LptF is an outer membrane protein that may be important for P. aeruginosa survival in harsh environments, including lung colonization in CF.

Lipotoxin F of Pseudomonas aeruginosa is an AlgU-dependent and alginate-independent outer membrane protein involved in resistance to oxidative stress and adhesion to A549 human lung epithelia

PubMed Central

Damron, F. Heath; Napper, Jennifer; Teter, M. Allison; Yu, Hongwei D.

2009-01-01

Chronic lung infection with P. aeruginosa and excessive neutrophil-associated inflammation are major causes of morbidity and mortality in patients with cystic fibrosis (CF). Overproduction of an exopolysaccharide known as alginate leads to the formation of mucoid biofilms that are resistant to antibiotics and host defences. Alginate overproduction or mucoidy is controlled by a stress-related ECF sigma factor AlgU/T. Mutation in the anti-sigma factor MucA is a known mechanism for conversion to mucoidy. Recently, we showed that inactivation of a kinase (KinB) in nonmucoid strain PAO1 results in overproduction of alginate. Here, we report the initial characterization of lipotoxin F (LptF, PA3692), an OmpA-like outer membrane protein that exhibited increased expression in the mucoid PAO1kinB mutant. The lipotoxin family of proteins has been previously shown to induce inflammation in lung epithelia, which may play a role in CF disease progression. Expression of LptF was observed to be AlgU-dependent and upregulated in CF isolates. Deletion of lptF from the kinB mutant had no effect on alginate production. Deletion of lptF from PAO1 caused a differential susceptibility to oxidants that can be generated by phagocytes. The lptF and algU mutants were more sensitive to hypochlorite than PAO1. However, the lptF mutant displayed increased resistance to hydrogen peroxide. LptF also contributed to adhesion to A549 human lung epithelial cells. Our data suggest that LptF is an outer membrane protein that may be important for P. aeruginosa survival in harsh environments, including lung colonization in CF. PMID:19332805

Immunization against diseases caused by Theileria parva: a review.

PubMed

Uilenberg, G

1999-09-01

Theileria parva is the causative agent of three epidemiologically different diseases, East Coast fever (ECF), Corridor disease and January disease, caused by 3 types of T. parva, T. p. parva, T. p. lawrencei and T. p. bovis, respectively. The history of immunization against these diseases has been marked by salient discoveries such as the immune status in recovered animals, the activity of tetracyclines during the incubation period, the possibility for cryopreserving supernatant of prefed ticks and the development of useful serological tests. The possibility of simultaneous administration of stabilate and long-acting tetracycline have greatly contributed to making the infection and treatment method operational. The importance of antigenic diversity in T. parva has been reflected in the difficulties related to the selection of the immunizing stock or combinations of stocks: a 'cocktail' of East African isolates may give broad protection against field challenge by ECF (T. parva parva), but Corridor disease is more problematic. On the other hand, certain single isolates may give equally good protection against ECF field challenge. Studies on the immunology of T. parva infection and the application of molecular tools have led to the discovery that sera of recovered animals neutralize sporozoites of various isolates, and to the p67 molecular vaccine; yet so far the only available method of immunizing against T. parva infections is the infection and treatment method or, in the case of T. parva bovis, the use of sublethal stabilate doses. Infection and treatment is applied on a fairly large scale in Zambia, and on a more limited scale in a few other countries. Immunity by this rather crude method is long-lasting and solid, but cross-immunity problems against some field strains remain. Furthermore, as immunized animals remain carriers, immunization may contribute to attaining and improving endemic stability in endemic areas in indigenous breeds with an adequate level of genetic tolerance to ECF. On the other hand, carrier animals may constitute a risk for spreading the disease into ECF-free areas where the vector is present. Other disadvantages of the method are that immunization of cattle during the incubation of naturally contracted East Coast fever will not prevent the disease and jeopardize its reputation. Furthermore, stabilates have to be cryopreserved, often a technical drawback, and contamination with undesirable pathogens may occur in tick-derived material. Therefore the need remains for the development of effective molecular vaccines and it must be remembered that immunization must be cost-effective and sustainable and it is only one aspect of integrated control of theileriosis and other tick-borne diseases. There is no universally valid strategy. Several factors have to be considered: value and susceptibility of cattle to theileriosis and to other tick-borne and tick-associated diseases, infestation by various ticks present in the area, the type of theileriosis (ECF, Corridor disease or January disease) and the epidemiological situation where immunization is taking place. The optimal age for immunization of the calves in endemic areas needs to be determined: when calf mortality by naturally occurring theileriosis is a problem, the sooner calves are immunized the better, but a proportion will have contracted natural infection before they can be reached, and immunization of very young calves might not be fully effective.

Assessing errors in the determination of base excess.

PubMed

Mentel, Alexander; Bach, Friedhelm; Schüler, Joerg; Herrmann, Walter; Koster, Andreas; Crystal, George J; Gatzounis, Georgios; Mertzlufft, Fritz

2002-05-01

We compared estimates for base excess of extracellular fluid (BE(ecf); mmol/L) obtained in five clinically used blood gas analyzers: AVL Compact 2 (Roche Diagnostics, Mannheim, Germany), Ciba-Corning 860 (Bayer Diagnostics, Fernwald, Germany), IL 1620 (Instrumentation Laboratories, Lexington, MA), Stat Profile Ultra (Nova Biomedical, Waltham, MA), and ABL 510 (Radiometer, Copenhagen, Denmark). A total of 134 measurements per analyzer were obtained in arterial and venous blood samples from 10 patients undergoing cardiac surgery and 65 measurements per analyzer in venous blood samples from 2 healthy volunteers. The blood samples were equilibrated in a tonometer with gases of known composition (37 degrees C). Additional theoretical studies were performed to evaluate the relationship between pH and calculated BE(ecf) value (with varied PCO(2)) using the formulas of the various analyzers. The standard deviations of repeated measurements were 0.24 mmol/L for ABL 510 and approximately 0.45 mmol/L for the other 4 analyzers. The maximal systematic difference between the average of all measurements of each analyzer was 3.7 mmol/L; this was primarily attributable to differences in measuring pH, and, to a lesser extent, to differences in calculation and determination of PCO(2). Comparison of the results from samples with different oxygen saturation showed that the relative alkalinity of deoxygenated hemoglobin (Haldane effect) can also influence the determinations of BE(ecf). A clinically useful way to quantify nonrespiratory disturbances of the acid-base balance is calculation of the base excess of extracellular fluid by using blood gas analyzers. In this study, we found significant variability in estimates of base excess of extracellular fluid obtained with five analyzers from different manufacturers. This variability is attributable to multiple factors, including lack of correction for deoxygenated hemoglobin (Haldane effect).

Alkaline peroxide treatment of ECF bleached softwood kraft pulps. Part 1. characterizing the effect of alkaline peroxide treatment on carboxyl groups of fibers

Treesearch

Zheng Dang; Thomas Elder; Arthur J. Ragauskas

2007-01-01

The influence of alkaline peroxide treatment has been characterized on elementally chlorine-free (ECF) bleached softwood (SW) kraft pulp. The results indicate that fiber charge increased with an increase in peroxide charge: a maximum fiber charge increment of 16.6% was obtained with 8.0% more peroxide charge on oven-dried (0.d.) pulp at 60.0°C. Two primary bleaching...

Evidence that the tri-cellular metabolism of N-acetylaspartate functions as the brain's "operating system": how NAA metabolism supports meaningful intercellular frequency-encoded communications.

PubMed

Baslow, Morris H

2010-11-01

N-acetylaspartate (NAA), an acetylated derivative of L-aspartate (Asp), and N-acetylaspartylglutamate (NAAG), a derivative of NAA and L-glutamate (Glu), are synthesized by neurons in brain. However, neurons cannot catabolize either of these substances, and so their metabolism requires the participation of two other cell types. Neurons release both NAA and NAAG to extra-cellular fluid (ECF) upon stimulation, where astrocytes, the target cells for NAAG, hydrolyze it releasing NAA back into ECF, and oligodendrocytes, the target cells for NAA, hydrolyze it releasing Asp to ECF for recycling to neurons. This sequence is unique as it is the only known amino acid metabolic cycle in brain that requires three cell types for its completion. The results of this cycling are two-fold. First, neuronal metabolic water is transported to ECF for its removal from brain. Second, the rate of neuronal activity is coupled with focal hyperemia, providing stimulated neurons with the energy required for transmission of meaningful frequency-encoded messages. In this paper, it is proposed that the tri-cellular metabolism of NAA functions as the "operating system" of the brain, and is essential for normal cognitive and motor activities. Evidence in support of this hypothesis is provided by the outcomes of two human inborn errors in NAA metabolism.

The biology of Theileria parva and control of East Coast fever - Current status and future trends.

PubMed

Nene, Vishvanath; Kiara, Henry; Lacasta, Anna; Pelle, Roger; Svitek, Nicholas; Steinaa, Lucilla

2016-06-01

Tremendous progress has been made over the last ten years on East Coast fever (ECF) research. Publication of a reference genome sequence of Theileria parva, the causative agent of ECF, has led to a more thorough characterization of the genotypic and antigenic diversity of the pathogen. It also facilitated identification of antigens that are targets of bovine major histocompatibility complex class I restricted cytotoxic T-lymphocytes (CTLs), induced by a live parasite-based infection and treatment method (ITM) vaccine. This has led to improved knowledge of epitope-specific T-cell responses to ITM that most likely contribute to the phenomenon of strain-specific immunity. The Muguga cocktail ITM vaccine, which provides broad-spectrum immunity to ECF is now a registered product in three countries in eastern Africa. Effort is directed at improving and scaling up the production process to make this vaccine more widely available on a commercial basis in the region. Meanwhile, research to develop a subunit vaccine based on parasite neutralizing antibodies and CTLs has been revived through convening of a research consortium to develop proof-of-concept for a next generation vaccine. Many new scientific and technical advances are facilitating this objective. Hence, the next decade promises even more progress toward an improved control of ECF. Copyright © 2016 The Authors. Published by Elsevier GmbH.. All rights reserved.

Inhibition and dispersal of Agrobacterium tumefaciens biofilms by a small diffusible Pseudomonas aeruginosa exoproduct(s)

PubMed Central

Hibbing, Michael E.; Fuqua, Clay

2013-01-01

Environmental biofilms often contain mixed populations of different species. In these dense communities, competition between biofilm residents for limited nutrients such as iron, can be fierce, leading to the evolution of competitive factors that affect the ability of competitors to grow or form biofilms. We have discovered a compound(s) present in the conditioned culture fluids of Pseudomonas aeruginosa that disperses and inhibits the formation of biofilms produced by the facultative plant pathogen Agrobacterium tumefaciens. The inhibitory activity is strongly induced when P. aeruginosa is cultivated in iron-limited conditions, but it does not function through iron sequestration. In addition, the production of the inhibitory activity is not regulated by the global iron regulatory protein Fur, the iron-responsive extra-cytoplasmic function (ECF) σ factor PvdS, or three of the recognized P. aeruginosa quorum sensing systems. In addition, the compound(s) responsible for the inhibition and dispersal of A. tumefaciens biofilm formation is likely distinct from the recently identified P. aeruginosa dispersal factor, cis-2-decenoic acid (CDA), as dialysis of the culture fluids showed that the inhibitory compound was larger than CDA and culture fluids that dispersed and inhibited biofilm formation by A. tumefaciens had no effect on biofilm formation by P. aeruginosa. PMID:22105093

Adjuvant Chemoradiation for Gastric Cancer Using Epirubicin, Cisplatin, and 5-Fluorouracil Before and After Three-Dimensional Conformal Radiotherapy With Concurrent Infusional 5-Fluorouracil: A Multicenter Study of the Trans-Tasman Radiation Oncology Group

DOE Office of Scientific and Technical Information (OSTI.GOV)

Leong, Trevor, E-mail: trevor.leong@petermac.or; Joon, Daryl Lim; Willis, David

Purpose: The INT0116 study has established postoperative chemoradiotherapy as the standard of care for completely resected gastric adenocarcinoma. However, the optimal chemoradiation regimen remains to be defined. We conducted a prospective, multicenter study to evaluate an alternative chemoradiation regimen that combines more current systemic treatment with modern techniques of radiotherapy delivery. Methods and Materials: Patients with adenocarcinoma of the stomach who had undergone an R0 resection were eligible. Adjuvant therapy consisted of one cycle of epirubicin, cisplatin, and 5-FU (ECF), followed by radiotherapy with concurrent infusional 5-FU, and then two additional cycles of ECF. Radiotherapy was delivered using precisely defined,more » multiple-field, three-dimensional conformal techniques. Results: A total of 54 assessable patients were enrolled from 19 institutions. The proportion of patients commencing Cycles 1, 2, and 3 of ECF chemotherapy were 100%, 81%, and 67% respectively. In all, 94% of patients who received radiotherapy completed treatment as planned. Grade 3/4 neutropenia occurred in 66% of patients with 7.4% developing febrile neutropenia. Most neutropenic episodes (83%) occurred in the post-radiotherapy period during cycles 2 and 3 of ECF. Grade 3/4 gastrointestinal toxicity occurred in 28% of patients. In all, 35% of radiotherapy treatment plans contained protocol deviations that were satisfactorily amended before commencement of treatment. At median follow-up of 36 months, the 3-year overall survival rate was estimated at 61.6%. Conclusions: This adjuvant regimen using ECF before and after three-dimensional conformal chemoradiation is feasible and can be safely delivered in a cooperative group setting. A regimen similar to this is currently being compared with the INT0116 regimen in a National Cancer Institute-sponsored, randomized Phase III trial.« less

Vitamin D status in relation to postural stability in the elderly.

PubMed

Boersma, D; Demontiero, O; Mohtasham Amiri, Z; Hassan, S; Suarez, H; Geisinger, D; Suriyaarachchi, P; Sharma, A; Duque, G

2012-03-01

Postural instability (PI) is an important risk factor for falls, especially in the frail older population. In this study, we investigated the impact of vitamin D deficiency on PI in a sample of community dwelling older subjects. Our objective was to determine the potential association between vitamin D deficiency and PI in older fallers. Cross-sectional study. Falls and Fractures Clinic, Department of Geriatric Medicine, Nepean Hospital, Penrith, Australia. One hundred and forty-five adults aged 65 years and older who have had at least one episode of a fall within the six months prior to assessment at the Falls and Fractures Clinic. Serum 25(OH) vitamin D3 [25(OH)D3] and parathyroid hormone concentrations were determined at baseline. Subjects were separated into 3 groups based on serum 25(OH)D3 levels with the following cut-off values: < 30 nmol/L (deficient), 30-50 nmol/L (insufficient) and > 50 nmol/L (normal). Other baseline measurements included body mass index, mini-nutritional assessment, grip strength, serum calcium concentration and creatinine clearance, which were used as covariables. PI was assessed using a computerized virtual reality system (Medicaa, Uruguay). Measured parameters included limits of stability (LOS) and centre of pressure (COP) under eyes closed on foam (ECF) and visio-vestibular stimulation. The estimated swaying area, computed from the ellipse of confidence under eyes closed standing on foam (ECF), was also used as a PI parameter. Gait velocity (GV) was measured using a GaitRITE walkway system. Posture was impaired in vitamin D deficiency (<30 nmol/L) as indicated by lower LOS (90 +/- 18), higher ECF (25 +/- 10) and slower GV (55 +/- 7) as compared with the insufficient and normal groups. After adjustment for demographic, biochemical and anthropometric variables, vitamin D deficiency significantly correlated with low LOS and high COP under ECF. Low levels of vitamin D were associated with PI. This association could also have an effect on slow GV and increased risk of falls. In conclusion, using an objective method to measure balance in older fallers we have identified a novel role of vitamin D in balance control. Prospective studies are required to confirm the effect of vitamin D on PI and elucidate the mechanisms of this association.

Novel baculovirus-derived p67 subunit vaccines efficacious against East Coast fever in cattle.

PubMed

Kaba, Stephen A; Musoke, Anthony J; Schaap, Dick; Schetters, Theo; Rowlands, John; Vermeulen, Arno N; Nene, Vishvanath; Vlak, Just M; van Oers, Monique M

2005-04-15

Two novel baculovirus-derived recombinant Theileria parva p67 constructs were tested for their vaccine potential against East Coast fever. Boran calves were immunized with a his-GFP-p67 fusion protein (GFP:p67deltaSS) or with GP64:p67C, a protein fusion between a C-terminal domain of p67 and the baculovirus envelope protein GP64. Both GFP:p67deltaSS and GP64:p67C induced antibodies with high ELISA titers that neutralized T. parva sporozoites with high efficiency. Upon challenge, a correlation was observed between the in vitro neutralizing capacity and the reduction in severe ECF for individual animals. A protection level upto 85% was obtained. This level of protection was achieved with only two inoculations of 100 microg per dose, which is a major improvement over previous recombinant p67 products.

In depth analysis of the mechanism of action of metal-dependent sigma factors: characterization of CorE2 from Myxococcus xanthus

PubMed Central

Marcos-Torres, Francisco Javier; Pérez, Juana; Gómez-Santos, Nuria; Moraleda-Muñoz, Aurelio; Muñoz-Dorado, José

2016-01-01

Extracytoplasmic function sigma factors represent the third pillar of signal-transduction mechanisms in bacteria. The variety of stimuli they recognize and mechanisms of action they use have allowed their classification into more than 50 groups. We have characterized CorE2 from Myxococcus xanthus, which belongs to group ECF44 and upregulates the expression of two genes when it is activated by cadmium and zinc. Sigma factors of this group contain a Cys-rich domain (CRD) at the C terminus which is essential for detecting metals. Point mutations at the six Cys residues of the CRD have revealed the contribution of each residue to CorE2 activity. Some of them are essential, while others are either dispensable or their mutations only slightly affect the activity of the protein. However, importantly, mutation of Cys174 completely shifts the specificity of CorE2 from cadmium to copper, indicating that the Cys arrangement of the CRD determines the metal specificity. Moreover, the conserved CxC motif located between the σ2 domain and the σ4.2 region has also been found to be essential for activity. The results presented here contribute to our understanding of the mechanism of action of metal-dependent sigma factors and help to define new common features of the members of this group of regulators. PMID:26951374

Association of the 1886 Charleston, South Carolina, earthquake and seismicity near Summervile with a 12º bend in the East Coast fault system and triple-fault junctions

USGS Publications Warehouse

Marple, R.; Miller, R.

2006-01-01

Seismic-reflection data were integrated with other geophysical, geologic, and seismicity data to better determine the location and nature of buried faults in the Charleston, South Carolina, region. Our results indicate that the 1886 Charleston, South Carolina, earthquake and seismicity near Summerville are related to local stresses caused by a 12?? bend in the East Coast fault system (ECFS) and two triple-fault junctions. One triple junction is formed by the intersection of the northwest-trending Ashley River fault with the two segments of the ECFS north and south of the bend. The other triple junction is formed by the intersection of the northeast-trending Summerville fault and a newly discovered northwest-trending Berkeley fault with the ECFS about 10 km north of the bend. The Summerville fault is a northwest-dipping border fault of the Triassic-age Jedburg basin that is undergoing reverse-style reactivation. This reverse-style reactivation is unusual because the Summerville fault parallels the regional stress field axis, suggesting that the reactivation is from stresses applied by dextral motion on the ECFS. The southwest-dip and reverse-type motion of the Berkeley fault are interpreted from seismicity data and a seismic-reflection profile in the western part of the study area. Our results also indicate that the East Coast fault system is a Paleozoic basement fault and that its reactivation since early Mesozoic time has fractured through the overlying allochthonous terranes.

Pharmacokinetic/Pharmacodynamic Relationship of Gabapentin in a CFA-induced Inflammatory Hyperalgesia Rat Model.

PubMed

Larsen, Malte Selch; Keizer, Ron; Munro, Gordon; Mørk, Arne; Holm, René; Savic, Rada; Kreilgaard, Mads

2016-05-01

Gabapentin displays non-linear drug disposition, which complicates dosing for optimal therapeutic effect. Thus, the current study was performed to elucidate the pharmacokinetic/pharmacodynamic (PKPD) relationship of gabapentin's effect on mechanical hypersensitivity in a rat model of CFA-induced inflammatory hyperalgesia. A semi-mechanistic population-based PKPD model was developed using nonlinear mixed-effects modelling, based on gabapentin plasma and brain extracellular fluid (ECF) time-concentration data and measurements of CFA-evoked mechanical hyperalgesia following administration of a range of gabapentin doses (oral and intravenous). The plasma/brain ECF concentration-time profiles of gabapentin were adequately described with a two-compartment plasma model with saturable intestinal absorption rate (K m  = 44.1 mg/kg, V max  = 41.9 mg/h∙kg) and dose-dependent oral bioavailability linked to brain ECF concentration through a transit compartment. Brain ECF concentration was directly linked to a sigmoid E max function describing reversal of hyperalgesia (EC 50, plasma  = 16.7 μg/mL, EC 50, brain  = 3.3 μg/mL). The proposed semi-mechanistic population-based PKPD model provides further knowledge into the understanding of gabapentin's non-linear pharmacokinetics and the link between plasma/brain disposition and anti-hyperalgesic effects. The model suggests that intestinal absorption is the primary source of non-linearity and that the investigated rat model provides reasonable predictions of clinically effective plasma concentrations for gabapentin.

Mycobacterium tuberculosis two-component systems and implications in novel vaccines and drugs.

PubMed

Zhou, PeiFu; Long, QuanXin; Zhou, YeXin; Wang, HongHai; Xie, JianPing

2012-01-01

Communication is vital for nearly all organisms to survive and thrive. For some particularly successful intracellular pathogens, a robust and precise signal transduction system is imperative for handling the complex, volatile, and harsh niche. The communication network of the etiology of tuberculosis, Mycobacterium tuberculosis (M.tb), namely two-component system (TCS), the eukaryotic-like Ser/Thr protein kinases(STPKs) system, the protein tyrosine kinase(PTK) system and the extracytoplasmic function σ(ECF-σ) system, determine how the pathogen responds to environmental fluctuations. At least 12 pair TCSs and four orphan proteins (three response regulators, Rv2884, Rv0260c, Rv0818, and one putative sensory transduction protein, Rv3143) can be found in the M.tb H37Rv genome. They regulate various aspects of M.tb, including virulence, dormancy, persistence, and drug resistance. This review focuses on the physiological roles of TCSs and the network of M.tb TCSs from a systems biology perspective. The implications of TCSs for better vaccine and new drug targets against tuberculosis are also examined.

Using participatory epidemiology to investigate management options and relative importance of tick-borne diseases amongst transhumant zebu cattle in Karamoja Region, Uganda.

PubMed

Byaruhanga, C; Oosthuizen, M C; Collins, N E; Knobel, D

2015-12-01

A participatory epidemiological (PE) study was conducted with livestock keepers in Moroto and Kotido districts, Karamoja Region, Uganda, between October and December 2013 to determine the management options and relative importance of tick-borne diseases (TBDs) amongst transhumant zebu cattle. Data collection involved 24 focus group discussions (each comprising 8-12 people) in 24 settlement areas (manyattas), key informant interviews (30), direct observation, a review of surveillance data, clinical examination, and laboratory confirmation of cases of TBDs. Methods used in group discussions included semi-structured interviews, simple ranking, pairwise ranking, matrix scoring, proportional piling and participatory mapping. The results of pairwise comparison showed the Ngakarimojong-named diseases, lokit (East Coast fever, ECF), lopid (anaplasmosis), loukoi (contagious bovine pleuropneumonia, CBPP), lokou (heartwater) and lokulam (babesiosis), were considered the most important cattle diseases in Moroto in that order, while ECF, anaplasmosis, trypanosomosis (ediit), CBPP and nonspecific diarrhoea (loleo) were most important in Kotido. Strong agreement between informant groups (Kendall's coefficient of concordance W=0.568 and 0.682; p<0.001) in pairwise ranking indicated that the diseases were a common problem in selected districts. East Coast fever had the highest median score for incidence (18% [range: 2, 33]) in Moroto, followed by anaplasmosis (17.5% [8,32]) and CBPP (9% [1,21]). Most animals that suffered from ECF, anaplasmosis, heartwater and babesiosis died, as the respective median scores for case fatality rates (CFR) were 89.5% (42, 100), 82.8% (63, 100), 66.7% (20, 100) and 85.7% (0, 100). In Kotido, diseases with high incidence scores were ECF (21% [6,32]), anaplasmosis (17% [10,33]) and trypanosomosis (8% [2,18]). The CFRs for ECF and anaplasmosis were 81.7% (44, 100) and 70.7% (48, 100), respectively. Matrix scoring revealed that disease indicators showed strong agreement (W=0.382-0.659, p<0.05-p<0.001) between informant groups. Inadequate knowledge, poor veterinary services and limited availability of drugs were the main constraints that hindered the control of TBDs. Hand picking of ticks was done by all pastoralists while hand spraying with acaricides was irregular, often determined by availability of drug supplies and money. It was concluded that TBDs, particularly ECF and anaplasmosis were important diseases in this pastoral region. Results from this study may assist in the design of feasible control strategies. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

Age-associated reduction of the count and functional activity of stromal precursor cells can be caused by both true reduction (exhaustion) of cell pool and regulatory effects of the organism.

PubMed

Gorskaya, Yu F; Danilova, T A; Nesterenko, V G

2011-06-01

The study was carried out on CBA mice using the method of heterotopic transplantation. A fragment of the femoral bone marrow (1/2) or spleen (1/5 of the organ) was transplanted under the renal capsule of a recipient. The following donor-recipient cross-transplantation variants were studied: young-young (Y-Y), young-old (Y-O), old-old (O-O), and old-young (O-Y). Cell suspensions were prepared from 2-month transplants inoculated in monolayer cultures and the cloning efficiency (ECF-F) of stromal precursor cells (CFC-F) was evaluated. The bone marrow transplant ECF-F and the count of CFC-F in the O-O group were 8-fold lower than in the Y-Y group. In the O-Y group, ECF-F was 3-fold higher than in the O-O group, but by 2.5 times lower than in the Y-Y group. ECF-F in Y-O group was 2-fold lower than in Y-Y group. The ECF-F and CFC-F count in spleen transplants in the O-O group were 4- and 6-fold lower, respectively, than in Y-Y group. However, in O-Y group ECF-F was 7-fold higher than in O-O group and higher than even in Y-Y group. The weight of induced ectopic bone tissue after transplantation of the osteoinductor (fragments of the allogenic urinary bladder mucosa) was 2-fold lower in the O-O vs. Y-Y group. However, comparison of the ectopic bone tissue weights in different experimental groups showed that osteoinductor activity of the bladder epithelium did not decrease, but increased 3-fold with age (O-Y:Y-Y). A 5-fold reduction of this proportion in groups where the osteoinductor was transplanted from old donors to old and young recipients (O-Y:O-O) could be attributed to age-specific reduction of the count of inducible osteogenic precursor cells (IOPC). The data in general suggest that age-specific reduction of the stromal precursor count and functional activity could be caused by the true reduction (exhaustion) of cell pool (bone marrow CFC-F; presumably, IOPC) and by the regulatory effects of the organism (bone marrow and splenic CFC-F, IOPC). These data seem to be significant for understanding of the role of osteogenic stromal precursor cells in the development of age-associated bone tissue defects, for example, senile osteoporosis.

Effects of synchronicity of carbohydrate and protein degradation on rumen fermentation characteristics and microbial protein synthesis.

PubMed

Seo, J K; Kim, M H; Yang, J Y; Kim, H J; Lee, C H; Kim, K H; Ha, Jong K

2013-03-01

A series of in vitro studies were carried out to determine i) the effects of enzyme and formaldehyde treatment on the degradation characteristics of carbohydrate and protein sources and on the synchronicity of these processes, and ii) the effects of synchronizing carbohydrate and protein supply on rumen fermentation and microbial protein synthesis (MPS) in in vitro experiments. Untreated corn (C) and enzyme-treated corn (EC) were combined with soy bean meal with (ES) and without (S) enzyme treatment or formaldehyde treatment (FS). Six experimental feeds (CS, CES, CFS, ECS, ECES and ECFS) with different synchrony indices were prepared. Highly synchronous diets had the greatest dry matter (DM) digestibility when untreated corn was used. However, the degree of synchronicity did not influence DM digestibility when EC was mixed with various soybean meals. At time points of 12 h and 24 h of incubation, EC-containing diets showed lower ammonia-N concentrations than those of C-containing diets, irrespective of the degree of synchronicity, indicating that more efficient utilization of ammonia-N for MPS was achieved by ruminal microorganisms when EC was offered as a carbohydrate source. Within C-containing treatments, the purine base concentration increased as the diets were more synchronized. This effect was not observed when EC was offered. There were significant effects on VFA concentration of both C and S treatments and their interactions. Similar to purine concentrations, total VFA production and individual VFA concentration in the groups containing EC as an energy source was higher than those of other groups (CS, CES and CFS). The results of the present study suggested that the availability of energy or the protein source are the most limiting factors for rumen fermentation and MPS, rather than the degree of synchronicity.

75 FR 70104 - Airworthiness Directives; Eurocopter France (ECF) Model SA330F, G, and J; and AS332C, L, L1, and...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-11-17

... Airworthiness Directives; Eurocopter France (ECF) Model SA330F, G, and J; and AS332C, L, L1, and L2 Helicopters... (ASB) No. 52.13 for the SA330F, G, and J helicopters, and 52.00.38 for the AS332C, C1, L, L1, and L2... authority citation for part 39 continues to read as follows: Authority: 49 U.S.C. 106(g), 40113, 44701. Sec...

Optimization of ECF bleaching of kraft pulp. Part 1, Optimal bleaching of hardwood pulps made with different alkali charges

Treesearch

Thomas J. McDonough; Shunichiro Uno; Alan W. Rudie; Charles E. Courchene

2008-01-01

In an earlier modeling study, we showed that when a hardwood kraft pulp is bleached in the D0(EO)D1ED2 sequence with a D0 stage kappa factor of 0.20, the brightness of the pulp emerging from the D2 stage can be accurately predicted from the brightness of the pulp entering that stage. The entering brightness, in turn is a well-defined function of the ratio of the D1...

Immunisation against Theileria parva in eastern Zambia: influence of maternal antibodies and demonstration of the carrier status.

PubMed

Marcotty, T; Brandt, J; Billiouw, M; Chaka, G; Losson, B; Berkvens, D

2002-12-11

Immunisation of calves by the infection and treatment method (I & T) has been extensively used in the eastern province of Zambia to control East Coast fever (ECF), a protozoan tick-borne disease. This paper presents the results of a field longitudinal study, which included a total of 148 Angoni calves. After immunisation against ECF, they were monitored for a full rainy season, coinciding with the main peak of activity of the vector of Theileria parva, the tick Rhipicephalus appendiculatus. Dysimmunisation (acute reaction generated by I & T immunisation), seroconversion and mortality are among the parameters recorded. The effect of maternal antibodies on these parameters was analysed and also studied in experimental conditions on two calves. Before immunisation, young calves had a higher seroprevalence than older animals (maternal antibodies) but their post-immunisation seroprevalence was lower. There was no evidence that their immunoprotection was weaker but this indicates that the post-immunisation seroconversion is probably not a reliable tool to monitor the efficacy of calf immunisation. The carrier state of cattle after immunisation was investigated in experimental conditions on three bovines whereas in the field, the infection prevalence in the ticks was estimated using the relation between the tick burden and the T. parva contacts with the calves. The ability of larval and nymphal R. appendiculatus ticks to pick-up T. parva from carriers and to transmit it to naïve animals after moulting was assessed. It was found that both instars are able to transmit clinical and lethal ECF but that the prevalence of T. parva infection in nymphs is much lower than in adults, confirming the primary role of adults in the transmission of ECF in endemic conditions. Similar results were obtained from the field whereby the ECF peak corresponds with the peak of adult R. appendiculatus activity. The infection prevalence in the ticks was however much lower in the field than in experimental conditions indicating that an important proportion of them feed on alternative hosts. Old ticks seemed to have lost part of their infectivity. Copyright 2002 Elsevier Science B.V.

A quantitative risk assessment of bovine theileriosis entering Luapula Province from Central Province in Zambia via live cattle imports from traditional and commercial production sectors.

PubMed

Makungu, C; Mwacalimba, K K

2014-09-01

Theileriosis or East Coast Fever (ECF) is an important livestock disease widespread in Zambia except for some provinces such as Luapula. This freedom status has been achieved due to strict livestock movement regulations that only authorise cattle imports from commercial farms implementing strict ECF control regimens. Recent increases in both the demand and price of beef in Zambia are stimulating a policy change towards a more inclusive inter-provincial trade in live cattle. This may also encourage the introduction of breeding cattle from high production pastoral sectors such as Central Province to stimulate the beef industry in disease free low production areas such as the Luapula Province. To estimate and compare the risks linked with those potential introductions of cattle from the traditional or commercial production sectors of the Central Province, a quantitative risk assessment model was developed. This risk comparison was necessary because the traditional livestock production sector accounts for over 79% of breeding cattle trade in Central Province but is characterised by minimalistic tick-borne disease control and a higher prevalence of ECF. We estimate that should the importation of breeding cattle from Central into Luapula Province be permitted, we could expect to import ECF by the introduction of infected animals at a median rate (5th and 95th percentiles) of every 0.44 years (0.12, 2.60), from the traditional sector compared to every 3.57 years (0.37, 103.6) from the commercial sector. Infected ticks would be expected to enter every 3.46 (0.66, 43.8) years via traditional cattle imports. These risks are strongly influenced by the prevalence of infection, performance of pre-transport screening tests, and the effectiveness of pre-transport tick cleansing. This assessment is expected to provide a model for tick borne disease risk assessments in similar settings, as well as inform ECF control, cattle trade, and stock movement policies in Zambia. Copyright © 2014 Elsevier B.V. All rights reserved.

Capture-recapture analysis of East Coast fever in smallholder dairy herds in the Dar es Salaam region of Tanzania.

PubMed

Kivaria, F M; Noordhuizen, J P T M

2010-05-01

The prevalence of and case fatality rate due to East Coast fever (ECF) were estimated in 1402 dairy cattle in 87 small herds in the Dar es Salaam region of Tanzania, from January 2003 to January 2005 using a capture-recapture method. Information on clinical cases and deaths due to ECF were obtained from farm records and from a questionnaire-based cross-sectional survey conducted between July 2003 and March 2005 as part of longitudinal studies of bovine mastitis in these herds. The number of clinical cases identified was 567 (from farm records) and 496 (from the questionnaire), and the number of deaths recorded were 305 (from farm records) and 251 (from the questionnaire). In all, 450 clinical cases and 191 deaths due to ECF were identified from the two sources, giving an observed prevalence of 32% (CI(95%) 30-35%) and observed case fatality rate of 42% (CI(95%) 38-47%). Following application of the capture-recapture method, the estimated number of clinical cases and deaths was 625 (CI(95%) 617-633) and 401 (CI(95%) 384-418), respectively. The respective prevalence and case fatality rates were 45% (CI(95%) 41-48%) and 64% (CI(95%) 60-68%). The estimates obtained using the capture-recapture method are higher than those identified by traditional cross-sectional studies conducted in the same study area, and probably provide a more accurate epidemiological picture of ECF in this region of Tanzania. Copyright 2009 Elsevier Ltd. All rights reserved.

Financial analysis of East Coast fever control strategies in traditionally managed Sanga cattle in Central Province of Zambia.

PubMed

Minjauw, B; Rushton, J; James, A D; Upton, M

1999-01-01

Five different East Coast fever (ECF)-control strategies (involving ECF immunisation by the infection-and-treatment method) were tested in groups of traditionally managed Sanga cattle in the Central Province of Zambia over a period of 2.5 years. Two groups were under intensive tick control (weekly spraying with acaricide)--one group immunised and the other non-immunised. Two groups were under no tick control--one group immunised and the other non-immunised. The fifth group was under seasonal tick control (18 sprays/year) and was immunised against ECF. The input and output data were used to construct discounted cash flows for each group. The seasonally sprayed and immunised group gave the highest net present value, and the non-immunised group with no tick control, the lowest. A break-even analysis showed that the immunisation costs could rise to US$25.9 per animal before profitability was affected. For herds under intensive tick control, immunisation was of no financial benefit. The results demonstrate the value of immunisation, and indicate the importance of its combination with seasonal tick-control measures.

Management of enterocutaneous fistulas.

PubMed

Schecter, William P

2011-06-01

Management of enterocutaneous fistulas (ECFs) involves (1) recognition and stabilization, (2) anatomic definition and decision, and (3) definitive operation. Phase 1 encompasses correction of fluid and electrolyte imbalance, skin protection, and nutritional support. Abdominal imaging defines the anatomy of the fistula in phase 2. ECFs that do not heal spontaneously require segmental resection of the bowel segment communicating with the fistula and restoration of intestinal continuity in phase 3. The enteroatmospheric fistula (EAF) is a malevolent condition requiring prolonged wound care and nutritional support. Complex abdominal wall reconstruction immediately following fistula resection is necessary for all EAFs. 2011 Elsevier Inc. All rights reserved.

Levels of heroin and its metabolites in blood and brain extracellular fluid after i.v. heroin administration to freely moving rats

PubMed Central

Gottås, A; Øiestad, E L; Boix, F; Vindenes, V; Ripel, Å; Thaulow, C H; Mørland, J

2013-01-01

BACKGROUND AND PURPOSE Heroin, with low affinity for μ-opioid receptors, has been considered to act as a prodrug. In order to study the pharmacokinetics of heroin and its active metabolites after i.v. administration, we gave a bolus injection of heroin to rats and measured the concentration of heroin and its metabolites in blood and brain extracellular fluid (ECF). EXPERIMENTAL APPROACH After an i.v. bolus injection of heroin to freely moving Sprague–Dawley rats, the concentrations of heroin and metabolites in blood samples from the vena jugularis and in microdialysis samples from striatal brain ECF were measured by ultraperformance LC-MS/MS. KEY RESULTS Heroin levels decreased very fast, both in blood and brain ECF, and could not be detected after 18 and 10 min respectively. 6-Monoacetylmorphine (6-MAM) increased very rapidly, reaching its maximal concentrations after 2.0 and 4.3 min, respectively, and falling thereafter. Morphine increased very slowly, reaching its maximal levels, which were six times lower than the highest 6-MAM concentrations, after 12.6 and 21.3 min, with a very slow decline during the rest of the experiment and only surpassing 6-MAM levels at least 30 min after injection. CONCLUSIONS AND IMPLICATIONS After an i.v. heroin injection, 6-MAM was the predominant opioid present shortly after injection and during the first 30 min, not only in the blood but also in rat brain ECF. 6-MAM might therefore mediate most of the effects observed shortly after heroin intake, and this finding questions the general assumption that morphine is the main and most important metabolite of heroin. PMID:23865556

Implementing a Pro-forma for Multidisciplinary Management of an Enterocutaneous Fistula: A Case Study.

PubMed

Samad, Sohel; Anele, Chukwuemeka; Akhtar, Mansoor; Doughan, Samer

2015-06-01

Optimal management of patients with an entercocutaneous fistula (ECF) requires utilization of the sepsis, nutrition, anatomy, and surgical procedure (SNAP) protocol. The protocol includes early detection and treatment of sepsis, optimizing patient nutrition through oral and parenteral routes, identifying the fistula anatomy, optimal fistula management, and proceeding to corrective surgery when appropriate. The protocol requires multidisciplinary team (MDT) coordination among surgeons, nurses, dietitians, stoma nurses, and physiotherapists. This case study describes a 70-year-old man who developed an ECF subsequent to a laparotomy for a small bowel obstruction. Following a period of ileus, 16 days post laparotomy the patient developed a high-output (2,000 mL per day) fistula. The patient also became pyrexial with raised inflammatory markers, requiring antibiotic treatment. Following development of his ECF, he was managed using the SNAP protocol for the duration of his admission; however, in implementing this protocol with this patient, clinicians noted fluid charts were inadequate to allow effective management of the variables. Thus, a new pro-forma was created that encompassed fluid balance, nutritional status, and pertinent blood test results, as well as perifistular skin condition, medication, and documentation of management plans from the MDT team. The pro-forma was recorded daily in the patient notes. Following implementation of the pro-forma and the SNAP protocol, the patient recovered well clinically over a period of 4 weeks with a decrease in his fistula output to 300-500 mL per day, and he was discharged with plans for further corrective surgery to resect the fistula and for bowel re-anastomoses. Although fluid charts are readily available, they do not include all pertinent variables for optimal management of patients with an ECF. Further research is needed to validate the pro-forma and evaluate its effect on patient outcomes.

Fixed-dose combination of losartan and hydrochlorothiazide significantly improves endothelial function in uncontrolled hypertension by low-dose amlodipine: a randomized study.

PubMed

Takase, Bonpei; Nagata, Masayoshi

2014-12-01

Flow-mediated dilation (FMD) and nitroglycerin-mediated dilation (NMD) in the brachial artery are well-known indices for evaluating endothelial function (ECF). The blood pressure-lowering effects of the combination of losartan (ARB) and low-dose hydrochlorothiazide (H: ARB-H; ARB, 50 mg and H, 12.5 mg) are useful. The aim of the present study was to examine whether the combination of losartan and low-dose hydrochlorothiazide could improve ECF. To investigate the effect of ARB-H on ECF in patients with uncontrolled hypertension despite the use of amlodipine (2.5 mg daily), we performed a randomized controlled open-labeled study by using the envelope method and assigned 42 patients to either a control (CTRL) group or an ARB-H combination group, both of which received amlodipine 2.5 mg daily during the treatment period. In addition, both the CTRL (n=21, 69±7 years old) and ARB-H groups (n=21, 69±7 years old) received additional behavioral modification. Before and after 8 weeks of therapy, FMD and NMD were measured in both groups using novel FMD equipment (UNEXEF18G). Although baseline FMD was not different between the two groups, post-therapy FMD increased in the ARB-H group (2.97±1.56 to 3.95±1.86%, p<0.05) but did not change significantly in the CTRL group (2.95±1.43 to 3.11±1.27%, NS). No significant change was seen in NMD when comparing baseline and post-therapy values in either group. No treatment complications were observed. A fixed-dose combination of losartan and hydrochlorothiazide enhances ECF, suggesting that this combination might have both anti-hypertensive and anti-atherosclerotic effects in patients with hypertension.

Preclinical examination of clofarabine in pediatric ependymoma: intratumoral concentrations insufficient to warrant further study.

PubMed

Patel, Yogesh T; Jacus, Megan O; Boulos, Nidal; Dapper, Jason D; Davis, Abigail D; Vuppala, Pradeep K; Freeman, Burgess B; Mohankumar, Kumarasamypet M; Throm, Stacy L; Gilbertson, Richard J; Stewart, Clinton F

2015-05-01

Clofarabine, a deoxyadenosine analog, was an active anticancer drug in our in vitro high-throughput screening against mouse ependymoma neurospheres. To characterize the clofarabine disposition in mice for further preclinical efficacy studies, we evaluated the plasma and central nervous system disposition in a mouse model of ependymoma. A plasma pharmacokinetic study of clofarabine (45 mg/kg, IP) was performed in CD1 nude mice bearing ependymoma to obtain initial plasma pharmacokinetic parameters. These estimates were used to derive D-optimal plasma sampling time points for cerebral microdialysis studies. A simulation of clofarabine pharmacokinetics in mice and pediatric patients suggested that a dosage of 30 mg/kg IP in mice would give exposures comparable to that in children at a dosage of 148 mg/m(2). Cerebral microdialysis was performed to study the tumor extracellular fluid (ECF) disposition of clofarabine (30 mg/kg, IP) in the ependymoma cortical allografts. Plasma and tumor ECF concentration-time data were analyzed using a nonlinear mixed effects modeling approach. The median unbound fraction of clofarabine in mouse plasma was 0.79. The unbound tumor to plasma partition coefficient (K pt,uu: ratio of tumor to plasma AUCu,0-inf) of clofarabine was 0.12 ± 0.05. The model-predicted mean tumor ECF clofarabine concentrations were below the in vitro 1-h IC50 (407 ng/mL) for ependymoma neurospheres. Thus, our results show the clofarabine exposure reached in the tumor ECF was below that associated with an antitumor effect in our in vitro washout study. Therefore, clofarabine was de-prioritized as an agent to treat ependymoma, and further preclinical studies were not pursued.

Preclinical examination of clofarabine in pediatric ependymoma: Intratumoral concentrations insufficient to warrant further study

PubMed Central

Patel, Yogesh T.; Jacus, Megan O.; Boulos, Nidal; Dapper, Jason D.; Davis, Abigail D.; Vuppala, Pradeep K.; Freeman, Burgess B.; Mohankumar, Kumarasamypet M.; Throm, Stacy L.; Gilbertson, Richard J.; Stewart, Clinton F.

2015-01-01

Clofarabine, a deoxyadenosine analog, was an active anticancer drug in our in vitro high-throughput screening against mouse ependymoma neurospheres. To characterize the clofarabine disposition in mice for further preclinical efficacy studies, we evaluated the plasma and central nervous system (CNS) disposition in a mouse model of ependymoma. A plasma pharmacokinetic study of clofarabine (45 mg/kg, IP) was performed in CD1 nude mice bearing ependymoma to obtain initial plasma pharmacokinetic parameters. These estimates were used to derive D-optimal plasma sampling time-points for cerebral microdialysis studies. A simulation of clofarabine pharmacokinetics in mice and pediatric patients suggested that a dosage of 30 mg/kg, IP in mice would give exposures comparable to that in children at a dosage of 148 mg/m2. Cerebral microdialysis was performed to study the tumor extracellular fluid (ECF) disposition of clofarabine (30 mg/kg, IP) in the ependymoma cortical allografts. Plasma and tumor ECF concentration-time data were analyzed using a nonlinear mixed effects modeling approach. The median unbound fraction of clofarabine in mouse plasma was 0.79. The unbound tumor to plasma partition coefficient (Kpt,uu: ratio of tumor to plasma AUCu,0-inf) of clofarabine was 0.12±0.05. The model predicted mean tumor ECF clofarabine concentrations were below the in vitro 1-hr IC50 (407 ng/mL) for ependymoma neurospheres. Thus, our results show the clofarabine exposure reached in the tumor ECF was below that associated with an antitumor effect in our in vitro washout study. Therefore, clofarabine was de-prioritized as an agent to treat ependymoma, and further preclinical studies were not pursued. PMID:25724157

Influence of peptide transporter 2 (PEPT2) on the distribution of cefadroxil in mouse brain: A microdialysis study.

PubMed

Chen, Xiaomei; Keep, Richard F; Liang, Yan; Zhu, Hao-Jie; Hammarlund-Udenaes, Margareta; Hu, Yongjun; Smith, David E

2017-05-01

Peptide transporter 2 (PEPT2) is a high-affinity low-capacity transporter belonging to the proton-coupled oligopeptide transporter family. Although many aspects of PEPT2 structure-function are known, including its localization in choroid plexus and neurons, its regional activity in brain, especially extracellular fluid (ECF), is uncertain. In this study, the pharmacokinetics and regional brain distribution of cefadroxil, a β-lactam antibiotic and PEPT2 substrate, were investigated in wildtype and Pept2 null mice using in vivo intracerebral microdialysis. Cefadroxil was infused intravenously over 4h at 0.15mg/min/kg, and samples obtained from plasma, brain ECF, cerebrospinal fluid (CSF) and brain tissue. A permeability-surface area experiment was also performed in which 0.15mg/min/kg cefadroxil was infused intravenously for 10min, and samples obtained from plasma and brain tissues. Our results showed that PEPT2 ablation significantly increased the brain ECF and CSF levels of cefadroxil (2- to 2.5-fold). In contrast, there were no significant differences between wildtype and Pept2 null mice in the amount of cefadroxil in brain cells. The unbound volume of distribution of cefadroxil in brain was 60% lower in Pept2 null mice indicating an uptake function for PEPT2 in brain cells. Finally, PEPT2 did not affect the influx clearance of cefadroxil, thereby, ruling out differences between the two genotypes in drug entry across the blood-brain barriers. These findings demonstrate, for the first time, the impact of PEPT2 on brain ECF as well as the known role of PEPT2 in removing peptide-like drugs, such as cefadroxil, from the CSF to blood. Copyright © 2017 Elsevier Inc. All rights reserved.

Contributions of the σ(W) , σ(M) and σ(X) regulons to the lantibiotic resistome of Bacillus subtilis.

PubMed

Kingston, Anthony W; Liao, Xiaojie; Helmann, John D

2013-11-01

In Bacillus subtilis, the extracytoplasmic function (ECF) σ factors σ(M) , σ(W) and σ(X) all contribute to resistance against lantibiotics. Nisin, a model lantibiotic, has a dual mode of action: it inhibits cell wall synthesis by binding lipid II, and this complex also forms pores in the cytoplasmic membrane. These activities can be separated in a nisin hinge-region variant (N20P M21P) that binds lipid II, but no longer permeabilizes membranes. The major contribution of σ(M) to nisin resistance is expression of ltaSa, encoding a stress-activated lipoteichoic acid synthase, and σ(X) functions primarily by activation of the dlt operon controlling d-alanylation of teichoic acids. Together, σ(M) and σ(X) regulate cell envelope structure to decrease access of nisin to its lipid II target. In contrast, σ(W) is principally involved in protection against membrane permeabilization as it provides little protection against the nisin hinge region variant. σ(W) contributes to nisin resistance by regulation of a signal peptide peptidase (SppA), phage shock proteins (PspA and YvlC, a PspC homologue) and tellurite resistance related proteins (YceGHI). These defensive mechanisms are also effective against other lantibiotics such as mersacidin, gallidermin and subtilin and comprise an important subset of the intrinsic antibiotic resistome of B. subtilis. © 2013 John Wiley & Sons Ltd.

Contributions of the σW, σM, and σX Regulons to the Lantibiotic Resistome of Bacillus subtilis

PubMed Central

Kingston, Anthony W.; Liao, Xiaojie; Helmann, John D.

2014-01-01

Summary In Bacillus subtilis, the extracytoplasmic function (ECF) σ factors σM, σW, and σX all contribute to resistance against lantibiotics. Nisin, a model lantibiotic, has a dual mode of action: it inhibits cell wall synthesis by binding lipid II, and this complex also forms pores in the cytoplasmic membrane. These activities can be separated in a nisin hinge-region variant (N20P M21P) that binds lipid II, but no longer permeabilizes membranes. The major contribution of σM to nisin resistance is expression of ltaSa, encoding a stress-activated lipoteichoic acid synthase, and σX functions primarily by activation of the dlt operon controlling D-alanylation of teichoic acids. Together, σM and σX regulate cell envelope structure to decrease access of nisin to its lipid II target. In contrast, σW is principally involved in protection against membrane permeabilization as it provides little protection against the nisin hinge region variant. σW contributes to nisin resistance by regulation of a signal peptide peptidase (SppA), phage shock proteins (PspA and YvlC, a PspC homolog), and tellurite resistance related proteins (YceGHI). These defensive mechanisms are also effective against other lantibiotics such as mersacidin, gallidermin, and subtilin and comprise an important subset of the intrinsic antibiotic resistome of B. subtilis. PMID:23980836

Theileriosis in Zambia: etiology, epidemiology and control measures.

PubMed

Nambota, A; Samui, K; Sugimoto, C; Kakuta, T; Onuma, M

1994-06-01

In Zambia, theileriosis manifests itself in the form of Corridor disease (CD), caused by Theileria parva lawrencei, and East Coast fever (ECF), caused by T. parva parva. Of the approximately 3 million cattle in Zambia, 1.4 million are at risk to theileriosis. ECF is found in the Northern and Eastern provinces of the country, while CD appears in Southern, Central, Lusaka and Copperbelt provinces. Theileriosis is a major constraint to the development of the livestock industry in Zambia, with losses of about 10,000 cattle per annum. The disease is spreading at a very fast rate, over-flowing its original borders. The epidemiology is complicated by, among other factors, the wide distribution of the tick vector, Rhipicephalus appendiculatus, which is found all over the country. The current strategy of relying on tick control and therapeutic drugs as a way of controlling the disease is becoming increasingly difficult for Zambia. This is because both curative drugs and acaricides are very costly. Immunization against theileriosis using the infection and treatment method as a way of controlling the disease is becoming increasingly accepted, provided local Theileria stocks are used. This paper reviews the incidence of theileriosis in the last 2 years, 1991 and 1992. It also gives a historical perspective of the disease, epidemiology and control measures presently in use.

Pharmacology of colloids and crystalloids.

PubMed

Griffel, M I; Kaufman, B S

1992-04-01

We have attempted to review body fluid distribution by compartments so that the reader understands the physiology of ICF and ECF, and the relationship between interstitial and intravascular fluids. Crystalloids such as NS and RL are distributed to the ECF, whereas colloids primarily remain intravascular for longer periods. Although effective, crystalloids tend to require larger volumes for infusion, and edema remains a problem. Colloids as a group are extremely effective volume expanders, but none is ideal. Albumin, hetastarch, dextran, and the less commonly used colloids each have significant toxicities that must be considered when using them. Intelligent choices can be made to optimize use of these fluids.

Feeding-associated alterations in striatal neurotransmitter release

NASA Technical Reports Server (NTRS)

Acworth, I. N.; Ressler, K.; Wurtman, R. J.

1989-01-01

Published evidence suggests a role for dopaminergic (DA) brain pathways in feeding-associated behaviors. Using the novel technique of brain microdialysis of striatal extracellular fluid (ECF) as an index of DA release, Church et al. described increases in levels of DA when animals had limited access to pellets, but not with free access. Dopamine release from the nucleus accumbens did increase with free access to pellets post starvation or after food reward. We used permanently implanted microdialysis probes to measure ECF levels of DA, DOPAC, HVA, and large neutral amino acids (LNAA) for up to 72 hours after implantation among rats experiencing different dietary regimens.

PACS/information systems interoperability using Enterprise Communication Framework.

PubMed

alSafadi, Y; Lord, W P; Mankovich, N J

1998-06-01

Interoperability among healthcare applications goes beyond connectivity to allow components to exchange structured information and work together in a predictable, coordinated fashion. To facilitate building an interoperability infrastructure, an Enterprise Communication Framework (ECF) was developed by the members of the Andover Working Group for Healthcare Interoperability (AWG-OHI). The ECF consists of four models: 1) Use Case Model, 2) Domain Information Model (DIM), 3) Interaction Model, and 4) Message Model. To realize this framework, a software component called the Enterprise Communicator (EC) is used. In this paper, we will demonstrate the use of the framework in interoperating a picture archiving and communication system (PACS) with a radiology information system (RIS).

Bone as an ion exchange system: evidence for a link between mechanotransduction and metabolic needs.

PubMed

Rubinacci, A; Covini, M; Bisogni, C; Villa, I; Galli, M; Palumbo, C; Ferretti, M; Muglia, M A; Marotti, G

2002-04-01

To detect whether the mutual interaction occurring between the osteocytes-bone lining cells system (OBLCS) and the bone extracellular fluid (BECF) is affected by load through a modification of the BECF-extracellular fluid (ECF; systemic extracellular fluid) gradient, mice metatarsal bones immersed in ECF were subjected ex vivo to a 2-min cyclic axial load of different amplitudes and frequencies. The electric (ionic) currents at the bone surface were measured by a vibrating probe after having exposed BECF to ECF through a transcortical hole. The application of different loads and different frequencies increased the ionic current in a dose-dependent manner. The postload current density subsequently decayed following an exponential pattern. Postload increment's amplitude and decay were dependent on bone viability. Dummy and static loads did not induce current density modifications. Because BECF is perturbed by loading, it is conceivable that OBLCS tends to restore BECF preload conditions by controlling ion fluxes at the bone-plasma interface to fulfill metabolic needs. Because the electric current reflects the integrated activity of OBLCS, its evaluation in transgenic mice engineered to possess genetic lesions in channels or matrix constituents could be helpful in the characterization of the mechanical and metabolic functions of bone.

Energy Consumption Forecasting Using Semantic-Based Genetic Programming with Local Search Optimizer.

PubMed

Castelli, Mauro; Trujillo, Leonardo; Vanneschi, Leonardo

2015-01-01

Energy consumption forecasting (ECF) is an important policy issue in today's economies. An accurate ECF has great benefits for electric utilities and both negative and positive errors lead to increased operating costs. The paper proposes a semantic based genetic programming framework to address the ECF problem. In particular, we propose a system that finds (quasi-)perfect solutions with high probability and that generates models able to produce near optimal predictions also on unseen data. The framework blends a recently developed version of genetic programming that integrates semantic genetic operators with a local search method. The main idea in combining semantic genetic programming and a local searcher is to couple the exploration ability of the former with the exploitation ability of the latter. Experimental results confirm the suitability of the proposed method in predicting the energy consumption. In particular, the system produces a lower error with respect to the existing state-of-the art techniques used on the same dataset. More importantly, this case study has shown that including a local searcher in the geometric semantic genetic programming system can speed up the search process and can result in fitter models that are able to produce an accurate forecasting also on unseen data.

Design of orthogonal genetic switches based on a crosstalk map of σs, anti-σs, and promoters

PubMed Central

Rhodius, Virgil A; Segall-Shapiro, Thomas H; Sharon, Brian D; Ghodasara, Amar; Orlova, Ekaterina; Tabakh, Hannah; Burkhardt, David H; Clancy, Kevin; Peterson, Todd C; Gross, Carol A; Voigt, Christopher A

2013-01-01

Cells react to their environment through gene regulatory networks. Network integrity requires minimization of undesired crosstalk between their biomolecules. Similar constraints also limit the use of regulators when building synthetic circuits for engineering applications. Here, we mapped the promoter specificities of extracytoplasmic function (ECF) σs as well as the specificity of their interaction with anti-σs. DNA synthesis was used to build 86 ECF σs (two from every subgroup), their promoters, and 62 anti-σs identified from the genomes of diverse bacteria. A subset of 20 σs and promoters were found to be highly orthogonal to each other. This set can be increased by combining the −35 and −10 binding domains from different subgroups to build chimeras that target sequences unrepresented in any subgroup. The orthogonal σs, anti-σs, and promoters were used to build synthetic genetic switches in Escherichia coli. This represents a genome-scale resource of the properties of ECF σs and a resource for synthetic biology, where this set of well-characterized regulatory parts will enable the construction of sophisticated gene expression programs. PMID:24169405

Electrodeposited manganese dioxide nanostructures on electro-etched carbon fibers: High performance materials for supercapacitor applications

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kazemi, Sayed Habib, E-mail: habibkazemi@iasbs.ac.ir; Center for Research in Climate Change and Global Warming; Maghami, Mostafa Ghaem

Highlights: • We report a facile method for fabrication of MnO{sub 2} nanostructures on electro-etched carbon fiber. • MnO{sub 2}-ECF electrode shows outstanding supercapacitive behavior even at high discharge rates. • Exceptional cycle stability was achieved for MnO{sub 2}-ECF electrode. • The coulombic efficiency of MnO{sub 2}-ECF electrode is nearly 100%. - Abstract: In this article we introduce a facile, low cost and additive/template free method to fabricate high-rate electrochemical capacitors. Manganese oxide nanostructures were electrodeposited on electro-etched carbon fiber substrate by applying a constant anodic current. Nanostructured MnO{sub 2} on electro-etched carbon fiber was characterized by scanning electron microscopy,more » X-ray diffraction and energy dispersive X-ray analysis. The electrochemical behavior of MnO{sub 2} electro-etched carbon fiber electrode was investigated by electrochemical techniques including cyclic voltammetry, galvanostatic charge/discharge, and electrochemical impedance spectroscopy. A maximum specific capacitance of 728.5 F g{sup −1} was achieved at a scan rate of 5 mV s{sup −1} for MnO{sub 2} electro-etched carbon fiber electrode. Also, this electrode showed exceptional cycle stability, suggesting that it can be considered as a good candidate for supercapacitor electrodes.« less

Development of an animal model of nephrocalcinosis via selective dietary sodium and chloride depletion

PubMed Central

Tuchman, Shamir; Asico, Laureano D.; Escano, Crisanto; Bobb, Daniel A.; Ray, Patricio E.

2013-01-01

Background Nephrocalcinosis (NC) is an important clinical problem seen in critically ill pre-term neonates treated with loop diuretics. No reliable animal models are available to study the pathogenesis of NC in preterm infants. The purpose of this study was to develop a reproducible and clinically relevant animal model of NC for these patients, and to explore the impact of extracellular fluid (ECF) volume contraction induced by sodium and chloride depletion in this process. Methods Three-week old weanling Sprague-Dawley rats were fed diets deficient in either chloride or sodium and chloride. A sub-group of rats from each dietary group was injected daily with furosemide (40 mg/kg; i.p.). Results Rats fed a control diet, with or without furosemide, or a chloride depleted diet alone, did not develop NC. In contrast, 50% of the rats injected with furosemide and fed the chloride depleted diet developed NC. Moreover, 94% of the rats fed the combined sodium/chloride depleted diet developed NC, independently of furosemide use. NC was associated with the development of severe ECF volume contraction, hypochloremic, hypokalemic metabolic alkalosis, increased phosphaturia, and growth retardation. Conclusion Severe ECF volume contraction induced by chronic sodium and chloride depletion appears to play an important role in the pathogenesis of NC. PMID:23174703

ECFS: A decentralized, distributed and fault-tolerant FUSE filesystem for the LHCb online farm

NASA Astrophysics Data System (ADS)

Rybczynski, Tomasz; Bonaccorsi, Enrico; Neufeld, Niko

2014-06-01

The LHCb experiment records millions of proton collisions every second, but only a fraction of them are useful for LHCb physics. In order to filter out the "bad events" a large farm of x86-servers (~2000 nodes) has been put in place. These servers boot from and run from NFS, however they use their local disk to temporarily store data, which cannot be processed in real-time ("data-deferring"). These events are subsequently processed, when there are no live-data coming in. The effective CPU power is thus greatly increased. This gain in CPU power depends critically on the availability of the local disks. For cost and power-reasons, mirroring (RAID-1) is not used, leading to a lot of operational headache with failing disks and disk-errors or server failures induced by faulty disks. To mitigate these problems and increase the reliability of the LHCb farm, while at same time keeping cost and power-consumption low, an extensive research and study of existing highly available and distributed file systems has been done. While many distributed file systems are providing reliability by "file replication", none of the evaluated ones supports erasure algorithms. A decentralised, distributed and fault-tolerant "write once read many" file system has been designed and implemented as a proof of concept providing fault tolerance without using expensive - in terms of disk space - file replication techniques and providing a unique namespace as a main goals. This paper describes the design and the implementation of the Erasure Codes File System (ECFS) and presents the specialised FUSE interface for Linux. Depending on the encoding algorithm ECFS will use a certain number of target directories as a backend to store the segments that compose the encoded data. When target directories are mounted via nfs/autofs - ECFS will act as a file-system over network/block-level raid over multiple servers.

Development of a new score to estimate clinical East Coast Fever in experimentally infected cattle.

PubMed

Schetters, Th P M; Arts, G; Niessen, R; Schaap, D

2010-02-10

East Coast Fever is a tick-transmitted disease in cattle caused by Theileria parva protozoan parasites. Quantification of the clinical disease can be done by determining a number of variables, derived from parasitological, haematological and rectal temperature measurements as described by Rowlands et al. (2000). From a total of 13 parameters a single ECF-score is calculated that allows categorization of infected cattle in five different classes that correlate with the severity of clinical signs. This score is complicated not only by the fact that it requires estimation of 13 parameters but also because of the subsequent mathematics. The fact that the values are normalised over a range of 0-10 for each experiment makes it impossible to compare results from different experiments. Here we present an alternative score based on the packed cell volume and the number of piroplasms in the circulation and that is calculated using a simple equation; ECF-score=PCV(relday0)/log(PE+10). In this equation the packed cell volume is expressed as a value relative to that of the day on infection (PCV(relday0)) and the number of piroplasms is expressed as the logarithmic value of the number of infected red blood cells (=PE) in a total of 1000 red blood cells. To allow PE to be 0, +10 is added in the denominator. We analysed a data set of 54 cattle from a previous experiment and found a statistically significant linear correlation between the ECF-score value reached during the post-infection period and the Rowlands' score value. The new score is much more practical than the Rowlands score as it only requires daily blood sampling. From these blood samples both PCV and number of piroplasms can be determined, and the score can be calculated daily. This allows monitoring the development of ECF after infection, which was hitherto not possible. In addition, the new score allows for easy comparison of results from different experiments.

A constitutively expressed pair of rpoE2-chrR2 in Azospirillum brasilense Sp7 is required for survival under antibiotic and oxidative stress.

PubMed

Gupta, Namrata; Kumar, Santosh; Mishra, Mukti Nath; Tripathi, Anil Kumar

2013-02-01

Extracytoplasmic function (ECF) sigma factors (σ(E)) are known to bring about changes in gene expression to enable bacteria to adapt to different stresses. The Azospirillum brasilense Sp245 genome harbours nine genes encoding σ(E), of which two are adjacent to the genes encoding ChrR-type zinc-binding anti-sigma (ZAS) factors. We describe here the role and regulation of a new pair of rpoE-chrR, which was found in the genome of A. brasilense Sp7 in addition to the previously described rpoE-chrR pair (designated rpoE1-chrR1). The rpoE2-chrR2 pair is also cotranscribed, and their products show protein-protein interaction. The -10 and -35 promoter elements of rpoE2-chrR2 and rpoE1-chrR1 were similar but not identical. Unlike the promoter of rpoE1-chrR1, the rpoE2-chrR2 promoter was neither autoregulated nor induced by oxidative stress. Inactivation of chrR2 or overexpression of rpoE2 in A. brasilense Sp7 resulted in an overproduction of carotenoids. It also conferred resistance to oxidative stresses and antibiotics. By controlling the synthesis of carotenoids, initiation and elongation of translation, protein folding and purine biosynthesis, RpoE2 seems to play a crucial role in preventing and repairing the cellular damage caused by oxidative stress. Lack of autoregulation and constitutive expression of rpoE2-chrR2 suggest that RpoE2-ChrR2 may provide a rapid mechanism to cope with oxidative stress, wherein singlet oxygen ((1)O(2))-mediated dissociation of the RpoE2-ChrR2 complex might release RpoE2 to drive the expression of its target genes.

Rate Coefficient Measurements and Theoretical Analysis of the OH + ( E)-CF3CH═CHCF3 Reaction.

PubMed

Baasandorj, Munkhbayar; Marshall, Paul; Waterland, Robert L; Ravishankara, A R; Burkholder, James B

2018-05-04

Rate coefficients, k, for the gas-phase reaction of the OH radical with ( E)-CF 3 CH═CHCF 3 (( E)-1,1,1,4,4,4-hexafluoro-2-butene, HFO-1336mzz(E)) were measured over a range of temperatures (211-374 K) and bath gas pressures (20-300 Torr; He, N 2 ) using a pulsed laser photolysis-laser-induced fluorescence (PLP-LIF) technique. k 1 ( T) was independent of pressure over this range of conditions with k 1 (296 K) = (1.31 ± 0.15) × 10 -13 cm 3 molecule -1 s -1 and k 1 ( T) = (6.94 ± 0.80) × 10 -13 exp[-(496 ± 10)/ T] cm 3 molecule -1 s -1 , where the uncertainties are 2σ, and the pre-exponential term includes estimated systematic error. Rate coefficients for the OD reaction were also determined over a range of temperatures (262-374 K) at 100 Torr (He). The OD rate coefficients were ∼15% greater than the OH values and showed similar temperature dependent behavior with k 2 ( T) = (7.52 ± 0.44) × 10 -13 exp[-(476 ± 20)/ T] and k 2 (296 K) = (1.53 ± 0.15) × 10 -13 cm 3 molecule -1 s -1 . The rate coefficients for reaction 1 were also measured using a relative rate technique between 296 and 375 K with k 1 (296 K) measured to be (1.22 ± 0.1) × 10 -13 cm 3 molecule -1 s -1 , in agreement with the PLP-LIF results. In addition, the 296 K rate coefficient for the O 3 + ( E)-CF 3 CH═CHCF 3 reaction was determined to be <5.2 × 10 -22 cm 3 molecule -1 s -1 . A theoretical computational analysis is presented to interpret the observed positive temperature dependence for the addition reaction and the significant decrease in OH reactivity compared to the ( Z)-CF 3 CH═CHCF 3 stereoisomer reaction. The estimated atmospheric lifetime of ( E)-CF 3 CH═CHCF 3 , due to loss by reaction with OH, is estimated to be ∼90 days, while the actual lifetime will depend on the location and season of its emission. Infrared absorption spectra of ( E)-CF 3 CH═CHCF 3 were measured and used to estimate the 100 year time horizon global warming potentials (GWP) of 32 (atmospherically well-mixed) and 14 (lifetime-adjusted).

Population pharmacokinetic modelling of non-linear brain distribution of morphine: influence of active saturable influx and P-glycoprotein mediated efflux

PubMed Central

Groenendaal, D; Freijer, J; de Mik, D; Bouw, M R; Danhof, M; de Lange, E C M

2007-01-01

Background and purpose: Biophase equilibration must be considered to gain insight into the mechanisms underlying the pharmacokinetic-pharmacodynamic (PK-PD) correlations of opioids. The objective was to characterise in a quantitative manner the non-linear distribution kinetics of morphine in brain. Experimental approach: Male rats received a 10-min infusion of 4 mg kg−1 of morphine, combined with a continuous infusion of the P-glycoprotein (Pgp) inhibitor GF120918 or vehicle, or 40 mg kg−1 morphine alone. Unbound extracellular fluid (ECF) concentrations obtained by intracerebral microdialysis and total blood concentrations were analysed using a population modelling approach. Key results: Blood pharmacokinetics of morphine was best described with a three-compartment model and was not influenced by GF120918. Non-linear distribution kinetics in brain ECF was observed with increasing dose. A one compartment distribution model was developed, with separate expressions for passive diffusion, active saturable influx and active efflux by Pgp. The passive diffusion rate constant was 0.0014 min−1. The active efflux rate constant decreased from 0.0195 min−1 to 0.0113 min−1 in the presence of GF120918. The active influx was insensitive to GF120918 and had a maximum transport (Nmax/Vecf) of 0.66 ng min−1 ml−1 and was saturated at low concentrations of morphine (C50=9.9 ng ml−1). Conclusions and implications: Brain distribution of morphine is determined by three factors: limited passive diffusion; active efflux, reduced by 42% by Pgp inhibition; low capacity active uptake. This implies blood concentration-dependency and sensitivity to drug-drug interactions. These factors should be taken into account in further investigations on PK-PD correlations of morphine. PMID:17471182

East Coast Fever Caused by Theileria parva Is Characterized by Macrophage Activation Associated with Vasculitis and Respiratory Failure

PubMed Central

Schneider, David A.; Frevert, Charles W.; Nelson, Danielle D.; Morrison, W. Ivan; Knowles, Donald P.

2016-01-01

Respiratory failure and death in East Coast Fever (ECF), a clinical syndrome of African cattle caused by the apicomplexan parasite Theileria parva, has historically been attributed to pulmonary infiltration by infected lymphocytes. However, immunohistochemical staining of tissue from T. parva infected cattle revealed large numbers of CD3- and CD20-negative intralesional mononuclear cells. Due to this finding, we hypothesized that macrophages play an important role in Theileria parva disease pathogenesis. Data presented here demonstrates that terminal ECF in both Holstein and Boran cattle is largely due to multisystemic histiocytic responses and resultant tissue damage. Furthermore, the combination of these histologic changes with the clinical findings, including lymphadenopathy, prolonged pyrexia, multi-lineage leukopenia, and thrombocytopenia is consistent with macrophage activation syndrome. All animals that succumbed to infection exhibited lymphohistiocytic vasculitis of small to medium caliber blood and lymphatic vessels. In pulmonary, lymphoid, splenic and hepatic tissues from Holstein cattle, the majority of intralesional macrophages were positive for CD163, and often expressed large amounts of IL-17. These data define a terminal ECF pathogenesis in which parasite-driven lymphoproliferation leads to secondary systemic macrophage activation syndrome, mononuclear vasculitis, pulmonary edema, respiratory failure and death. The accompanying macrophage phenotype defined by CD163 and IL-17 is presented in the context of this pathogenesis. PMID:27195791

East Coast Fever Caused by Theileria parva Is Characterized by Macrophage Activation Associated with Vasculitis and Respiratory Failure.

PubMed

Fry, Lindsay M; Schneider, David A; Frevert, Charles W; Nelson, Danielle D; Morrison, W Ivan; Knowles, Donald P

2016-01-01

Respiratory failure and death in East Coast Fever (ECF), a clinical syndrome of African cattle caused by the apicomplexan parasite Theileria parva, has historically been attributed to pulmonary infiltration by infected lymphocytes. However, immunohistochemical staining of tissue from T. parva infected cattle revealed large numbers of CD3- and CD20-negative intralesional mononuclear cells. Due to this finding, we hypothesized that macrophages play an important role in Theileria parva disease pathogenesis. Data presented here demonstrates that terminal ECF in both Holstein and Boran cattle is largely due to multisystemic histiocytic responses and resultant tissue damage. Furthermore, the combination of these histologic changes with the clinical findings, including lymphadenopathy, prolonged pyrexia, multi-lineage leukopenia, and thrombocytopenia is consistent with macrophage activation syndrome. All animals that succumbed to infection exhibited lymphohistiocytic vasculitis of small to medium caliber blood and lymphatic vessels. In pulmonary, lymphoid, splenic and hepatic tissues from Holstein cattle, the majority of intralesional macrophages were positive for CD163, and often expressed large amounts of IL-17. These data define a terminal ECF pathogenesis in which parasite-driven lymphoproliferation leads to secondary systemic macrophage activation syndrome, mononuclear vasculitis, pulmonary edema, respiratory failure and death. The accompanying macrophage phenotype defined by CD163 and IL-17 is presented in the context of this pathogenesis.

Construction of exchange repulsion in terms of the wave functions at QM/MM boundary region

DOE Office of Scientific and Technical Information (OSTI.GOV)

Takahashi, Hideaki, E-mail: hideaki@m.tohoku.ac.jp; Umino, Satoru; Morita, Akihiro

2015-08-28

We developed a simple method to calculate exchange repulsion between a quantum mechanical (QM) solute and a molecular mechanical (MM) molecule in the QM/MM approach. In our method, the size parameter in the Buckingham type potential for the QM solute is directly determined in terms of the one-electron wave functions of the solute. The point of the method lies in the introduction of the exchange core function (ECF) defined as a Slater function which mimics the behavior of the exterior electron density at the QM/MM boundary region. In the present paper, the ECF was constructed in terms of the Becke-Rousselmore » (BR) exchange hole function. It was demonstrated that the ECF yielded by the BR procedure can faithfully reproduce the radial behavior of the electron density of a QM solute. The size parameter of the solute as well as the exchange repulsion are, then, obtained using the overlap model without any fitting procedure. To examine the efficiency of the method, it was applied to calculation of the exchange repulsions for minimal QM/MM systems, hydrogen-bonded water dimer, and H{sub 3}O{sup +}–H{sub 2}O. We found that our approach is able to reproduce the potential energy curves for these systems showing reasonable agreements with those given by accurate full quantum chemical calculations.« less

Cystic fibrosis research topics featured at the 14th ECFS Basic Science Conference: Chairman's summary.

PubMed

Mall, Marcus A; Hwang, Tzyh-Chang; Braakman, Ineke

2018-03-01

In recent years, tremendous progress has been made in the development of novel drugs targeting the basic defect in patients with cystic fibrosis (CF). This breakthrough is based on a solid foundation of knowledge on CFTR's function in health and how mutations in CFTR cause CF multi-organ disease. This knowledge has been collected and continuously expanded by an active and persistent CF research community and has paved the way for precision medicine for CF. Since 2004, the European Cystic Fibrosis Society (ECFS) has held an annual Basic Science Conference that has evolved as an international forum for interdisciplinary discussion of hot topics and unsolved questions related to CF research. This Special Issue reviews CF research topics featured at the 14th ECFS Basic Science Conference and provides an up-to-date overview of recent progress in our understanding of CFTR structure and function, disease mechanisms implicated in airway mucus plugging, inflammation and abnormal host-pathogen interactions, and advancements with enhanced cell and animal model systems and breakthrough therapies directed at mutant CFTR or alternative targets. In addition, this Special Issue also identifies a number of fundamental questions and hurdles that still have to be overcome to realize the full potential of precision medicine and develop transformative therapies for all patients with CF. Copyright © 2017 European Cystic Fibrosis Society. Published by Elsevier B.V. All rights reserved.

Complex regulation of AprA metalloprotease in Pseudomonas fluorescens M114: evidence for the involvement of iron, the ECF sigma factor, PbrA and pseudobactin M114 siderophore.

PubMed

Maunsell, Bláithín; Adams, Claire; O'Gara, Fergal

2006-01-01

In the soil bacterium Pseudomonas fluorescens M114, extracellular proteolytic activity and fluorescent siderophore (pseudobactin M114) production were previously shown to be co-ordinately negatively regulated in response to environmental iron levels. An iron-starvation extracytoplasmic function sigma factor, PbrA, required for the transcription of siderophore biosynthetic genes, was also implicated in M114 protease regulation. The current study centred on the characterization and genetic regulation of the gene(s) responsible for protease production in M114. A serralysin-type metalloprotease gene, aprA, was identified and found to encode the major, if not only, extracellular protease produced by this strain. The expression of aprA and its protein product were found to be subject to complex regulation. Transcription analysis confirmed that PbrA was required for full aprA transcription under low iron conditions, while the ferric uptake regulator, Fur, was implicated in aprA repression under high iron conditions. Interestingly, the iron regulation of AprA was dependent on culture conditions, with PbrA-independent AprA-mediated proteolytic activity observed on skim milk agar supplemented with yeast extract, when supplied with iron or purified pseudobactin M114. These effects were not observed on skim milk agar without yeast extract. PbrA-independent aprA expression was also observed from a truncated transcriptional fusion when grown in sucrose asparagine tryptone broth supplied with iron or purified pseudobactin M114. Thus, experimental evidence suggested that iron mediated its effects via transcriptional activation by PbrA under low iron conditions, while an as-yet-unidentified sigma factor(s) may be required for the PbrA-independent aprA expression and AprA proteolytic activity induced by siderophore and iron.

Ependymin as a substrate for outgrowth of axons from cultured explants of goldfish retina.

PubMed

Schmidt, J T; Schmidt, R; Lin, W C; Jian, X Y; Stuermer, C A

1991-01-01

Ependymin, a prominent protein of the brain's extracellular fluid (ECF) was previously implicated in the consolidation of memory and in the activity-driven sharpening of the retinotectal projection. Because both these phenomena probably involve the growth and elaboration of appropriate synapses, we have tested whether ependymin can serve as a substrate for the growth of axons from goldfish retinal ganglion cells in a culture assay. Ependymin (Ep), laminin (LAM), polylysine (PL), and Concanavalin A (Con A) were plated on glass coverslips either uniformly or in striped patterns. Ep alone, either soluble or partly polymerized (by dropping calcium concentration and pH), was a good substrate for axonal outgrowth, as good or better than PL and Con A, but not as good as LAM. Neurites grew faster on LAM (71 microns/h) than on Ep (32 microns/h) or on PL (22 microns/h). Fasciculation was low on LAM, intermediate on Ep, and highest on PL. In exclusive side-by-side stripe assays, axons preferred LAM over Ep, but gave weak or no preference for Ep over Con A or PL. With stripes of LAM + Ep alongside pure LAM, the axons preferred the mixture of LAM + Ep. When antibodies to Ep were plated in stripes over continuous Ep substrate, the axons avoided the antibody-blocked stripes and grew on the Ep stripes. Antibodies to Ep did not, however, block growth on laminin substrates, nor did antibodies to LAM block growth on Ep. Dot blots and western blots showed very little cross recognition between the antibodies. Ependymin is a good substrate for neurite outgrowth, which is normally present in ECF, and adhesion to Ep is independent of LAM and possibly additive to it.

Effect of agro-ecological zone and grazing system on incidence of East Coast Fever in calves in Mbale and Sironko Districts of Eastern Uganda.

PubMed

Rubaire-Akiiki, Christopher M; Okello-Onen, Joseph; Musunga, David; Kabagambe, Edmond K; Vaarst, Mettee; Okello, David; Opolot, Charles; Bisagaya, A; Okori, C; Bisagati, C; Ongyera, S; Mwayi, M T

2006-08-17

Between May 2002 and February 2003 a longitudinal survey was carried out in Mbale and Sironko Districts of Eastern Uganda to determine the influence of agro-ecological zones (AEZ) and grazing systems on tick infestation patterns and incidence of East Coast Fever (ECF) in bovine calves. The study area was stratified into AEZ (lowland, midland and upland) and grazing systems {zero grazing (ZG), restricted-outdoor grazing (ROG) and communal grazing (CG)}, whose strata had previously been shown to influence the prevalence of ECF, babesiosis and anaplasmosis. One hundred and eighty-five smallholder dairy farms with a total of 198 calves of both sexes, between the ages of 1 day and 6 weeks, were purposively selected from the AEZ-grazing system strata. Nine dynamic cohorts (11-51 calves in each) of these calves were examined and sampled monthly. Ticks infesting the calves were counted from one side of the animal body and categorized into the different species, sex and feeding status. Sera were collected at recruitment and monthly thereafter and antibodies against Theileria parva, T. mutans, Babesia bigemina, B. bovis and Anaplasma marginale were measured using ELISA. Tick challenge (total and specific) varied with AEZ and grazing system. The risk of infection with T. parva was higher in the lowland zone compared to the upland zone (hazard ratio (HR)=2.59; 95% CI: 1.00-6.34). The risk of infection with T. parva was higher in the CG system than the ZG system (HR=10.00; 95% CI: 3.61-27.92). The incidence risk for sero-conversion, over the 10 months study period, was 62, 16 and 9% in the lowland, midland and upland zones, respectively. Ninety-eight percent of the calves in lowland-CG stratum sero-converted by the age of 6 months, while 56 and 8% did so in the lowland-ROG and the lowland-ZG stratum, respectively. The results of this study show the need to consider farm circumstances and the variation in ECF risk, both spatially and temporally when designing control strategies for ECF.

Modelling the delay between pharmacokinetics and EEG effects of morphine in rats: binding kinetic versus effect compartment models.

PubMed

de Witte, Wilhelmus E A; Rottschäfer, Vivi; Danhof, Meindert; van der Graaf, Piet H; Peletier, Lambertus A; de Lange, Elizabeth C M

2018-05-18

Drug-target binding kinetics (as determined by association and dissociation rate constants, k on and k off ) can be an important determinant of the kinetics of drug action. However, the effect compartment model is used most frequently instead of a target binding model to describe hysteresis. Here we investigate when the drug-target binding model should be used in lieu of the effect compartment model. The utility of the effect compartment (EC), the target binding kinetics (TB) and the combined effect compartment-target binding kinetics (EC-TB) model were tested on either plasma (EC PL , TB PL and EC-TB PL ) or brain extracellular fluid (ECF) (EC ECF , TB ECF and EC-TB ECF ) morphine concentrations and EEG amplitude in rats. It was also analyzed when a significant shift in the time to maximal target occupancy (Tmax TO ) with increasing dose, the discriminating feature between the TB and EC model, occurs in the TB model. All TB models assumed a linear relationship between target occupancy and drug effect on the EEG amplitude. All three model types performed similarly in describing the morphine pharmacodynamics data, although the EC model provided the best statistical result. The analysis of the shift in Tmax TO (∆Tmax TO ) as a result of increasing dose revealed that ∆Tmax TO is decreasing towards zero if the k off is much smaller than the elimination rate constant or if the target concentration is larger than the initial morphine concentration. The results for the morphine PKPD modelling and the analysis of ∆Tmax TO indicate that the EC and TB models do not necessarily lead to different drug effect versus time curves for different doses if a delay between drug concentrations and drug effect (hysteresis) is described. Drawing mechanistic conclusions from successfully fitting one of these two models should therefore be avoided. Since the TB model can be informed by in vitro measurements of k on and k off , a target binding model should be considered more often for mechanistic modelling purposes.

Time-frequency analyses of fluid-solid interaction under sinusoidal translational shear deformation of the viscoelastic rat cerebrum

NASA Astrophysics Data System (ADS)

Leahy, Lauren N.; Haslach, Henry W.

2018-02-01

During normal extracellular fluid (ECF) flow in the brain glymphatic system or during pathological flow induced by trauma resulting from impacts and blast waves, ECF-solid matter interactions result from sinusoidal shear waves in the brain and cranial arterial tissue, both heterogeneous biological tissues with high fluid content. The flow in the glymphatic system is known to be forced by pulsations of the cranial arteries at about 1 Hz. The experimental shear stress response to sinusoidal translational shear deformation at 1 Hz and 25% strain amplitude and either 0% or 33% compression is compared for rat cerebrum and bovine aortic tissue. Time-frequency analyses aim to correlate the shear stress signal frequency components over time with the behavior of brain tissue constituents to identify the physical source of the shear nonlinear viscoelastic response. Discrete fast Fourier transformation analysis and the novel application to the shear stress signal of harmonic wavelet decomposition both show significant 1 Hz and 3 Hz components. The 3 Hz component in brain tissue, whose magnitude is much larger than in aortic tissue, may result from interstitial fluid induced drag forces. The harmonic wavelet decomposition locates 3 Hz harmonics whose magnitudes decrease on subsequent cycles perhaps because of bond breaking that results in easier fluid movement. Both tissues exhibit transient shear stress softening similar to the Mullins effect in rubber. The form of a new mathematical model for the drag force produced by ECF-solid matter interactions captures the third harmonic seen experimentally.

Influence of organic matter on arsenic removal by continuous flow electrocoagulation treatment of weakly mineralized waters.

PubMed

Pallier, Virginie; Feuillade-Cathalifaud, Geneviève; Serpaud, Bernard

2011-03-01

The aim of this study is to evaluate and understand the electrocoagulation/flocculation (ECF) process to remove arsenic from both model and natural waters with low mineral content and to compare its performances to the coagulation/flocculation (CF) process already optimized. Experiments were thus conducted with iron electrodes in the same specific treatment conditions (4≤current density (mAcm(-2))≤33) to study the influence of organic matter on arsenic removal in conditions avoiding the oxidation step usually required to improve As(III) removal. The process performance was evaluated by combining quantification of arsenic residual concentrations and speciation and dissolved organic carbon residual concentrations with zeta potential and turbidity measurements. When compared to CF, ECF presented several disadvantages: (i) lower As(V) removal yield because of the ferrous iron dissolved from the anode and the subsequent negative zeta potential of the colloidal suspension, (ii) higher residual DOC concentrations because of the fractionation of high molecular weight compounds during the treatment leading to compounds less prone to coagulate and (iii) higher residual turbidities because of the charge neutralization mechanisms involved. However, during this process, As(III) was oxidized to As(V) improving considerably its removal whatever the matrix conditions. ECF thus allowed to improve As(III) removal without applying an oxidation step that could potentially lead to the formation of toxic oxidation by-products. Copyright © 2011 Elsevier Ltd. All rights reserved.

Relationships between coastal bacterioplankton growth rates and biomass production: comparison of leucine and thymidine uptake with single-cell physiological characteristics.

PubMed

Franco-Vidal, Leticia; Morán, Xosé Anxelu G

2011-02-01

Specific growth rates of heterotrophic bacterioplankton have been frequently estimated from in situ bacterial production (BP) to biomass (BB) ratios, using a series of assumptions that may result in serious discrepancies with values obtained from predator-free cultures. Here, we used both types of approaches together with a comprehensive assessment of single-cell physiological characteristics (membrane integrity, nucleic acid content, and active respiration) of coastal bacterioplankton during a complete annual cycle (February 2007-January 2008) in the southern Bay of Biscay off Xixón, Spain. Both leucine and thymidine incorporation rates were used in conjunction with empirical tracer to carbon or cells conversion factors (eCFs) to accurately derive BP. Leu and TdR incorporation rates covaried year-round, as did the corresponding eCFs at 0 and 50 m depth. eCFs peaked in autumn, with mean annual values close to the theoretical ones (3.4 kg C mol Leu(-1) and 2.0 × 10(18) cells mol TdR(-1)). Bacterial abundance (0.2-1.5 × 10(6) cells L(-1)) showed a bimodal distribution with maxima in May and October and minima in March. Live (membrane-intact) cells dominated year-round (79-97%), with high nucleic acid cells (42-88%) and actively respiring bacteria (CTC+, 1-16%) showing distinct surface maxima in April and July, respectively. BB (557-1,558 mg C m(-2)) and BP (7-139 mg C m(-2) day(-1)) presented two distinct peaks in spring and autumn, both of similar size due to a strong upwelling event observed in September. Specific growth rates (0.35-3.8 day(-1)) were one order of magnitude higher in predator-free incubations than bacterial turnover rates derived from integrated BP:BB ratios (0.01-0.16 and 0.01-0.09 day(-1), for Leu and TdR, respectively) and were not correlated, probably due to a significant contribution of low activity cells to total standing stocks. The Leu:TdR molar ratio averaged for the water column (6.6-25.5) decreased significantly with higher integrated BB, indicating that low standing stocks tend to present unbalanced growth. Discrepancies about the true magnitude of specific growth rates must be solved before fully appreciating the role of bacteria in the ocean carbon cycle.

Diversity in ABC transporters: Type I, II and III importers

PubMed Central

Rice, Austin J.; Park, Aekyung

2014-01-01

ATP-binding cassette transporters are multi-subunit membrane pumps that transport substrates across membranes. While significant in the transport process, transporter architecture exhibits a range of diversity that we are only beginning to recognize. This divergence may provide insight into the mechanisms of substrate transport and homeostasis. Until recently, ABC importers have been classified into two types, but with the emergence of energy-coupling factor (ECF) transporters there are potentially three types of ABC importers. In this review, we summarize an expansive body of research on the three types of importers with an emphasis on the basics that underlie ABC importers, such as structure, subunit composition and mechanism. PMID:25155087

Factors Affecting the Inter-annual to Centennial Time Scale Variability of All Indian Summer Monsoon Rainfall

NASA Astrophysics Data System (ADS)

Malik, Abdul; Brönnimann, Stefan

2016-04-01

The All Indian Summer Monsoon Rainfall (AISMR) is highly important for the livelihood of more than 1 billion people living in the Indian sub-continent. The agriculture of this region is heavily dependent on seasonal (JJAS) monsoon rainfall. An early start or a slight delay of monsoon, or an early withdrawal or prolonged monsoon season may upset the farmer's agricultural plans, can cause significant reduction in crop yield, and hence economic loss. Understanding of AISMR is also vital because it is a part of global atmospheric circulation system. Several studies show that AISMR is influenced by internal climate forcings (ICFs) viz. ENSO, AMO, PDO etc. as well as external climate forcings (ECFs) viz. Greenhouse Gases, volcanic eruptions, and Total Solar Irradiance (TSI). We investigate the influence of ICFs and ECFs on AISMR using recently developed statistical technique called De-trended Partial-Cross-Correlation Analysis (DPCCA). DPCCA can analyse a complex system of several interlinked variables. Often, climatic variables, being cross correlated, are simultaneously tele-connected with several other variables and it is not easy to isolate their intrinsic relationship. In the presence of non-stationarities and background signals the calculated correlation coefficients can be overestimated and erroneous. DPCCA method removes the non-stationarities and partials out the influence of background signals from the variables being cross correlated and thus give a robust estimate of correlation. We have performed the analysis using NOAA Reconstructed SSTs and homogenised instrumental AISMR data set from 1854-1999. By employing the DPCCA method we find that there is a statistically insignificant negative intrinsic relation (by excluding the influence of ICFs, and ECFs except TSI) between AISMR and TSI on decadal to centennial time scale. The ICFs considerably modulate the relation between AISMR and solar activity between 50-80 year time scales and transform this relationship to statistically significant positive. We conclude that the positive relation between AISMR and solar activity, as found by other authors, is due to the combined effect of AMO, PDO and multi-decadal ENSO variability on AISMR. The solar activity influences the ICFs and this influence is then transmitted to AISMR. Further, we find that there is statistically positive intrinsic relation between AISMR and AMO from 26 to 100 year time scales which is modulated by ICFs (PDO, ENSO) and ECFs. PDO, ENSO, and solar activity weaken this intrinsic relationship whereas the combined effect of ECFc (solar activity, volcanic eruptions, CO2, & tropospheric aerosol optical depth) results in strengthening of this relationship from 70 to 100 year time scales. There is a negative intrinsic relation between AISMR and PDO which is not statistically significant at any time scale. However this relationship becomes statistically significant only in the presence of combined effect of North Atlantic SSTs and ENSO (4-39 year time scale) and individual effect of TSI (3-26 year time scale) on AISMR. We also find that there is statistical significant negative relationship between AISMR and ENSO on inter-annual to centennial time scale and the strength of this relationship is modulated by solar activity from 3 to 40 year time scale.

Extracolonic findings (ECF) on CT colonography (CTC) in patients presenting with colorectal symptoms.

PubMed

Badiani, Sarit; Tomas-Hernandez, Silvia; Karandikar, Sharad; Roy-Choudhury, Shuvro

2013-10-01

Computed tomographic colonography (CTC) is now an established method for imaging the colon and rectum in the screening and symptomatic setting. Additional benefit of CTC is the ability to assess for extracolonic findings especially in patients presenting with colorectal symptoms. To determine prevalence of extracolonic findings (ECF) in symptomatic patients undergoing CTC and determine accuracy of CTC for exclusion of significant abdominal disease and extracolonic malignancy (ECM). A total of 1359 unenhanced prone and postcontrast supine CTC studies were performed between March 2002 and December 2007. ECF were retrospectively classified according to C-RADS criteria into E1 to E4 findings. For ECM, a gold standard of clinical and/or radiological follow-up supplemented with data from the regional cancer registry with a median follow-up of 42 months was created. Sensitivity and negative predictive values for ECM was calculated. Following exclusions, 1177 CTCs were analyzed. Of 1423 extracolonic findings reported, 328/1423 (23%) E3 and 100/1423 (7%) E4 (including six eventual FP studies) findings were identified. Thirty-two ECMs were confirmed following further investigations. Seven further small ECMs were detected during the entire follow-up, of which two were potentially visible in retrospect (false-negative studies). Additional tests were generated from 55/1177 (4.7%) studies. Sensitivity and negative predictive value for ECM was 94.1% (95% CI 78.9-98.9%) and 99.8% (95% CI 99.3-99.9%), respectively. One in 37 patients were found to have an ECM. Two potentially detectable cancers were missed. Only a small proportion of patients underwent additional work-up.

Exploring knowledge and management practices on ticks and tick-borne diseases among agro-pastoral communities in Southern Highlands, Tanzania.

PubMed

Kerario, Isack Ibrahim; Simuunza, Martin; Laisser, Emmanuel L K; Chenyambuga, Sebastian

2018-01-01

The current study was conducted to assess the farmers' knowledge and management practices on ticks and tick-borne diseases (TBDs) through individual interview using a structured questionnaire in Mbarali and Momba districts of Mbeya region. A total of 240 households, 120 from each district were asked to mention TBDs of cattle which they thought were the most important in their localities and period of the year when the diseases occurred more frequently. In addition, farmers were asked to describe clinical signs and management practices associated with the common TBDs that they knew. The majority of respondents (46.2%) reported that East Coast fever (ECF) was the most important disease of cattle in the region, followed by anaplasmosis (33.8%), heartwater (15.4%), and babesiosis (4.6%). According to the farmers, ECF and anaplasmosis occurred more frequently during the dry season, while babesiosis and heartwater occurred more frequently during the rainy season. The majority of farmers were able to describe properly the signs of the common TBDs. Most farmers (80.4%) reported that they used acaricide to control ticks at a frequency of after every 2 weeks and a small proportion (15.8%) vaccinated their animals against ECF. It can be concluded that farmers in Mbeya have considerable knowledge on tick species and clinical signs of TBDs affecting their cattle. Based on the findings of the current study, it is recommended that integrated approach to the control of ticks and TBDs be adopted in the study area and many other areas that utilize agro-pastoral and pastoral cattle production systems.

Governing equations for electro-conjugate fluid flow

NASA Astrophysics Data System (ADS)

Hosoda, K.; Takemura, K.; Fukagata, K.; Yokota, S.; Edamura, K.

2013-12-01

An electro-conjugation fluid (ECF) is a kind of dielectric liquid, which generates a powerful flow when high DC voltage is applied with tiny electrodes. This study deals with the derivation of the governing equations for electro-conjugate fluid flow based on the Korteweg-Helmholtz (KH) equation which represents the force in dielectric liquid subjected to high DC voltage. The governing equations consist of the Gauss's law, charge conservation with charge recombination, the KH equation, the continuity equation and the incompressible Navier-Stokes equations. The KH equation consists of coulomb force, dielectric constant gradient force and electrostriction force. The governing equation gives the distribution of electric field, charge density and flow velocity. In this study, direct numerical simulation (DNS) is used in order to get these distribution at arbitrary time. Successive over-relaxation (SOR) method is used in analyzing Gauss's law and constrained interpolation pseudo-particle (CIP) method is used in analyzing charge conservation with charge recombination. The third order Runge-Kutta method and conservative second-order-accurate finite difference method is used in analyzing the Navier-Stokes equations with the KH equation. This study also deals with the measurement of ECF ow generated with a symmetrical pole electrodes pair which are made of 0.3 mm diameter piano wire. Working fluid is FF-1EHA2 which is an ECF family. The flow is observed from the both electrodes, i.e., the flow collides in between the electrodes. The governing equation successfully calculates mean flow velocity in between the collector pole electrode and the colliding region by the numerical simulation.

Aetiology, management and outcome of entero-cutaneous fistula in Maiduguri, Nigeria.

PubMed

Eni, U E; Na'aya, H U; Gali, B M

2007-03-01

Enterocutaneous fistula (ECF) remains an important surgical problem with significant morbidity and mortality. This study aims to review the aetiology and management outcome in a depressed economy like ours. A retrospective review of 54 patients with ECF admitted into the surgical wards of the University of Maiduguri Teaching Hospital (UMTH) between January 1994 and December 2004 (11 year period). Thirty two (59.3%) were males and 22 (40.7%) were females giving a male/female ratio of 1.5 to 1. The age ranged from 1 to 58 years with two peak incidents of 20-29 years and 40-49 years. Eighteen cases (33%) occurred following appendicectomy, 12 (22%) following laparotomy for intestinal obstruction, 10 (18.5%) following laparotomy for abdominal malignancies, 7 (13%) followed laparotomy for penetrating abdominal injuries, 3 (5.5%) followed laparotomy for perforated typhoid enteritis, 2 (3.7%) cases were due to spontaneous rupture of strangulated and neglected inguinal hernia, 1 (1.9%) case followed chest tube insertion for pleural effusion in a PTB patient and 1 (1.9%) case followed a native healer's incision on a lumber hernia. Altogether,45 (83.3%) were referred cases from peripheral hospitals. Fourty one (76%) were high output type, while 13 (24%) were low output type. Most patients 32 (59.3%) healed spontaneously on conservative management. Eighteen (33%) had surgical intervention. Eight patients demised giving a mortality rate of 15%. The average hospital stay was 56 days. The main cause of ECF in our environment is postoperative (94.4%) with post appendicectomy cases alone accounting for 33%. Majority of our patients (66.7%) were managed conservatively.

Ba, B, and U element partitioning in magnesian calcite skeletons of Octocorallia corals

NASA Astrophysics Data System (ADS)

Yoshimura, T.; Suzuki, A.; Iwasaki, N.

2015-01-01

Barium, boron and uranium element partitioning and oxygen and carbon isotope fractionation of high-Mg calcite skeletons of Octocorallia corals were investigated. The dissolved Ba concentration in seawater and the coral Ba/Ca ratio showed a clear positive correlation. The empirically derived barium partition coefficient is comparable to previous data for not only calcitic corals but also intermediate- to deep-water-dwelling scleractinian corals whose skeletons are composed of aragonite. Octocorallia corals are geologically important producers of biominerals, and they provide long-term records (up to hundreds of years) of environmental conditions in the deep ocean. Our data suggest that Ba/Ca ratios in Octocorallia corals may be a useful proxy for nutrients in intermediate and deep waters. The Ba/Ca ratio, a possible proxy for pH or carbonate ion concentration in seawater, showed the largest correlation with δ13C among the examined parameters. This result implies that the pH of the extracytoplasmic calcifying fluid (ECF) simultaneously influences δ18O, δ13C, and Ba/Ca by influencing the relative contributions of dissolved carbon sources in the ECF. Positive correlations of Ba/Ca with δ18 and δ13C suggest that δ18 and δ13C are enriched in light isotopes when conditions are less alkaline, suggesting a potential role of biological alkalinity pumping becomes more favorable with decreasing calcifying fluid pH. Substantial inter- and intra-specimen variations in Ba/Ca suggest that physicochemical factors do not exert a dominant systematic control on U incorporation.

Functional Characterization of Corynebacterium glutamicum Mycothiol S-Conjugate Amidase

PubMed Central

Si, Meiru; Long, Mingxiu; Chaudhry, Muhammad Tausif; Xu, Yixiang; Zhang, Pan; Zhang, Lei; Shen, Xihui

2014-01-01

The present study focuses on the genetic and biochemical characterization of mycothiol S-conjugate amidase (Mca) of Corynebacterium glutamicum. Recombinant C. glutamicum Mca was heterologously expressed in Escherichia coli and purified to apparent homogeneity. The molecular weight of native Mca protein determined by gel filtration chromatography was 35 kDa, indicating that Mca exists as monomers in the purification condition. Mca showed amidase activity with mycothiol S-conjugate of monobromobimane (MSmB) in vivo while mca mutant lost the ability to cleave MSmB. In addition, Mca showed limited deacetylase activity with N-acetyl-D-glucosamine (GlcNAc) as substrate. Optimum pH for amidase activity was between 7.5 and 8.5, while the highest activity in the presence of Zn2+ confirmed Mca as a zinc metalloprotein. Amino acid residues conserved among Mca family members were located in C. glutamicum Mca and site-directed mutagenesis of these residues indicated that Asp14, Tyr137, His139 and Asp141 were important for activity. The mca deletion mutant showed decreased resistance to antibiotics, alkylating agents, oxidants and heavy metals, and these sensitive phenotypes were recovered in the complementary strain to a great extent. The physiological roles of Mca in resistance to various toxins were further supported by the induced expression of Mca in C. glutamicum under various stress conditions, directly under the control of the stress-responsive extracytoplasmic function-sigma (ECF-σ) factor SigH. PMID:25514023

75 FR 10692 - Television Broadcasting Services; Birmingham, AL

Federal Register 2010, 2011, 2012, 2013, 2014

2010-03-09

... FEDERAL COMMUNICATIONS COMMISSION 47 CFR Part 73 [DA 10-340; MB Docket No. 10-21; RM-11590] Television Broadcasting Services; Birmingham, AL AGENCY: Federal Communications Commission. ACTION: Final... ( http://fjallfoss.fcc.gov/ecfs/ ). This document may be purchased from the Commission's duplicating...

75 FR 1546 - Television Broadcasting Services; Bangor, ME

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2010-01-12

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2010-04-16

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76 FR 48789 - Adoption of Recommendations

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2011-08-09

..., contractor ethics, and video hearings. FOR FURTHER INFORMATION CONTACT: For Recommendation 2011-1, Emily... hosting public meetings online or using social media, blogs, and other web applications to promote public... Communications Commission, which has its own electronic comment filing system ( http://fjallfoss.fcc.gov/ecfs...

Extracellular calcium sensing and extracellular calcium signaling

NASA Technical Reports Server (NTRS)

Brown, E. M.; MacLeod, R. J.; O'Malley, B. W. (Principal Investigator)

2001-01-01

The cloning of a G protein-coupled extracellular Ca(2+) (Ca(o)(2+))-sensing receptor (CaR) has elucidated the molecular basis for many of the previously recognized effects of Ca(o)(2+) on tissues that maintain systemic Ca(o)(2+) homeostasis, especially parathyroid chief cells and several cells in the kidney. The availability of the cloned CaR enabled the development of DNA and antibody probes for identifying the CaR's mRNA and protein, respectively, within these and other tissues. It also permitted the identification of human diseases resulting from inactivating or activating mutations of the CaR gene and the subsequent generation of mice with targeted disruption of the CaR gene. The characteristic alterations in parathyroid and renal function in these patients and in the mice with "knockout" of the CaR gene have provided valuable information on the CaR's physiological roles in these tissues participating in mineral ion homeostasis. Nevertheless, relatively little is known about how the CaR regulates other tissues involved in systemic Ca(o)(2+) homeostasis, particularly bone and intestine. Moreover, there is evidence that additional Ca(o)(2+) sensors may exist in bone cells that mediate some or even all of the known effects of Ca(o)(2+) on these cells. Even more remains to be learned about the CaR's function in the rapidly growing list of cells that express it but are uninvolved in systemic Ca(o)(2+) metabolism. Available data suggest that the receptor serves numerous roles outside of systemic mineral ion homeostasis, ranging from the regulation of hormonal secretion and the activities of various ion channels to the longer term control of gene expression, programmed cell death (apoptosis), and cellular proliferation. In some cases, the CaR on these "nonhomeostatic" cells responds to local changes in Ca(o)(2+) taking place within compartments of the extracellular fluid (ECF) that communicate with the outside environment (e.g., the gastrointestinal tract). In others, localized changes in Ca(o)(2+) within the ECF can originate from several mechanisms, including fluxes of calcium ions into or out of cellular or extracellular stores or across epithelium that absorb or secrete Ca(2+). In any event, the CaR and other receptors/sensors for Ca(o)(2+) and probably for other extracellular ions represent versatile regulators of numerous cellular functions and may serve as important therapeutic targets.

Coagulation-flocculation as pre-treatment for micro-scale Fe/Cu/O3 process (CF-mFe/Cu/O3) treatment of the coating wastewater from automobile manufacturing.

PubMed

Xiong, Zhaokun; Cao, Jinyan; Yang, Dan; Lai, Bo; Yang, Ping

2017-01-01

A coagulation-flocculation as pre-treatment combined with mFe/Cu/O 3 (CF-mFe/Cu/O 3 ) process was developed to degrade the pollutants in automobile coating wastewater (ACW). In coagulation-flocculation (CF) process, high turbidity removal efficiency (97.1%) and low COD removal efficiency (10.5%) were obtained under the optimal conditions using Al 2 (SO 4 ) 3 ·18H 2 O and CaO. The effluent of CF process (ECF) was further disposed by mFe/Cu/O 3 process, and its key operating parameters were optimized by batch experiments. Optimally, COD removal efficiency of ECF obtained by the mFe/Cu/O 3 process (i.e., 87.6% after 30 min treatment) was much higher than those of mFe/Cu alone (8.3%), ozone alone (46.6%), and mFe/Cu/air (6.1%), which confirms the superiority of the mFe/Cu/O 3 process. In addition, the analysis results of UV-vis, excitation-emission matrix (EEM) fluorescence spectra and GC/MS further confirm that the phenol pollutants of ECF had been effectively decomposed or transformed after CF-mFe/Cu/O 3 process treatment. Meanwhile, B/C ratio of ACW increased from 0.19 to 0.56, which suggests the biodegradability was improved significantly. Finally, the operating cost of CF-mFe/Cu/O 3 process was about 1.83 USD t -1 for ACW treatment. Therefore, the combined process is a promising treatment technology for the coating wastewater from automobile manufacturing. Copyright © 2016 Elsevier Ltd. All rights reserved.

DUODENECTOMY: MANAGING THE FISTULA.

PubMed

Govender, M; Matsevych, O; Ghoor, F; Singh, N; Molenaar, C

2017-09-01

Duodenectomy is rarely indicated, however, in certain circumstances may have be performed. Enterocutaneous fistula (ECF) is the main cause of serious adverse outcomes. Its management remains challenging. Two cases of emergency duodenectomy are presented. The management of ECF is described and discussed. Case 1: A 22-year-old male presented in septic shock with perforated duodenal ulcer, suffered two cardiac arrests before index surgery. During re-laparotomy for leak, mobilisation resulted in an extensive injury of the duodenum extending to the ampula. A drainage procedure with complete duodenectomy and gastrojejenostomy was performed. The bile and pancreatic ducts were cannulated with infant feeding catheters and were separately pumped in the gastrostomy with a feeding pump. In one week, the patient had oral intake in addition to infused feeds. He remained in hospital for six months, suffered six episodes of gram-negative sepsis requiring antibiotics. Three episodes resulted from blocked catheters and cholangitis. Other were central line sepsis. A reconstruction with separate limbs of jejunum to the bile and pancreatic ducts was performed. He was well at 18 months post final surgery. Case 2: A 63-year-old male presented with a perforated hepatic flexure colonic carcinoma. Intraoperatively, the tumour invaded and partly obstructed the duodenum which was injured during mobilisation. A duodenectomy with the right hemicolectomy was performed. The jejunum was anastomosed to the ampulla. Leaking effluent from a drain was pumped into the gastrostomy. He was discharged in three weeks and continued to transfer the effluent himself with a 60 ml syringe until the leak ceased. Three months post-surgery, he developed metastasis to the drain tract and died in two months. Duodenectomy is feasible but complications are difficult to manage. ECF is common and should be managed by pumping the effluent in the gastrostomy until definitive surgery or spontaneous closure of the fistula.

Immunisation against East Coast fever by the infection and treatment method: evaluation of the use of ice baths for field delivery and appraisal of an acid formulation of long-acting tetracycline.

PubMed

Marcotty, T; Billiouw, M; Chaka, G; Berkvens, D; Losson, B; Brandt, J

2001-08-20

Immunisation by the infection and treatment method using the Katete strain is currently the most efficient prophylactic technique to control East Coast fever (ECF) in the endemic areas of the Eastern Province of Zambia. The maintenance of the cold chain in liquid nitrogen up to the time of inoculation and the cost of the reference long-acting oxytetracycline (Terramycin LA, Pfizer) are the main drawbacks of the method. The work presented in this paper aims at reducing the cost of immunisation against ECF by using an ice bath for the field delivery and a cheaper long-acting oxytetracycline formulation as chemotherapeutic agent. In experimental conditions, the results from 40 calves immunised after various periods of storage on ice ranging from 4 to 32 h indicate that deferred immunisation performed with a stabilate kept on ice for up to 6h after thawing has an efficiency of 90%. Moreover, sporozoites kept on ice were still surviving 32 h after thawing. In a field trial, 91 calves were inoculated with a stabilate kept for 3.5-5.5 h after thawing and dilution whereas 86 calves were immunised using the standard method. Clinical and parasitological reactions to immunisation were monitored as well as the seroconversion. In the field trial, the deferred immunisation was more efficient than the standard method. The acid formulation of oxytetracycline that was tested was found as suitable as the reference alkaline formulation for the chemotherapeutic control of the Katete strain in ECF immunisation. One indoor trial was carried out on 10 animals and a field trial involved 93 calves.

Exploring knowledge and management practices on ticks and tick-borne diseases among agro-pastoral communities in Southern Highlands, Tanzania

PubMed Central

Kerario, Isack Ibrahim; Simuunza, Martin; Laisser, Emmanuel L. K.; Chenyambuga, Sebastian

2018-01-01

Aim: The current study was conducted to assess the farmers’ knowledge and management practices on ticks and tick-borne diseases (TBDs) through individual interview using a structured questionnaire in Mbarali and Momba districts of Mbeya region. Materials and Methods: A total of 240 households, 120 from each district were asked to mention TBDs of cattle which they thought were the most important in their localities and period of the year when the diseases occurred more frequently. In addition, farmers were asked to describe clinical signs and management practices associated with the common TBDs that they knew. Results: The majority of respondents (46.2%) reported that East Coast fever (ECF) was the most important disease of cattle in the region, followed by anaplasmosis (33.8%), heartwater (15.4%), and babesiosis (4.6%). According to the farmers, ECF and anaplasmosis occurred more frequently during the dry season, while babesiosis and heartwater occurred more frequently during the rainy season. The majority of farmers were able to describe properly the signs of the common TBDs. Most farmers (80.4%) reported that they used acaricide to control ticks at a frequency of after every 2 weeks and a small proportion (15.8%) vaccinated their animals against ECF. Conclusion: It can be concluded that farmers in Mbeya have considerable knowledge on tick species and clinical signs of TBDs affecting their cattle. Based on the findings of the current study, it is recommended that integrated approach to the control of ticks and TBDs be adopted in the study area and many other areas that utilize agro-pastoral and pastoral cattle production systems. PMID:29479157

The Influence of Exogenous Fat and Water on Lumbar Spine Bone Mineral Density in Healthy Volunteers

PubMed Central

Kim, Kyu-Nam; Kim, Bom-Taeck; Kim, Kwang-Min; Park, Sat-Byul; Joo, Nam-Seok; Je, Sang Hyeon; Kim, Young-Sang

2012-01-01

Purpose Changes in human body composition can affect the accuracy of spine bone mineral density (BMD) measurements. The purpose of this study was to evaluate whether fat and water in the soft tissue of the abdomen influence lumbar spine BMD measurements obtained using dual energy X-ray absorptiometry (DXA). Materials and Methods Duplicate BMD measurements were carried out on healthy volunteers (10 men and 10 women) and the Hologic anthropomorphic spine phantom had on the same day before and after placement of following 3 materials in the abdominal area: lard 900 g, 1.5 cm thick; oil 1.4 liters in a vinyl bag; and water 1.2 liters in a vinyl bag. Results In the case of human participants, following the placement of exogenous water to mimic extracellular fluid (ECF), there was a significant decrease in lumbar spine BMD (-0.012 g/cm2, p=0.006), whereas the placement of exogenous lard and oil to mimic abdominal fat produced a slight increase in lumbar spine BMD (0.006 g/cm2, p=0.301; 0.008 g/cm2, p=0.250, respectively). The average percentage of lumbar spine BMD change with and without exogenous lard, oil, and water showed increase of 0.51%, and 0.67%, and decrease of 1.02%, respectively. Using the phantom, BMD decreased with the placement of both lard (-0.002 g/cm2, p=0.699) and water (-0.006 g/cm2, p=0.153); however, there was no difference in BMD after oil placement. Conclusion These results suggest that in cases where changes in fat and ECF volume are similar, ECF exerts a greater influence than fat on DXA lumbar BMD measurements. PMID:22318815

77 FR 64446 - Wireless Microphones Proceeding

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2012-10-22

... FEDERAL COMMUNICATIONS COMMISSION 47 CFR Parts 15, 74, and 90 [WT Docket Nos. 08-166, 08-167, ET Docket No. 10-24; DA 12-1570] Wireless Microphones Proceeding AGENCY: Federal Communications Commission.... [ssquf] Federal Communications Commission's Web site: http://www.fcc.gov/cgb/ecfs2/ . Follow the...

77 FR 16470 - Implementation of the Commercial Spectrum Enhancement Act and Modernization of the Commission's...

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2012-03-21

... purchased from the Commission's duplicating contractor, Best Copy and Printing, Inc. (BCPI), 445 12th Street.../ecfs/ . I. Background 1. In Council Tree Communications, Inc. v. FCC, 619 F.3d 235 (3d Cir. 2010), cert...

78 FR 78809 - Rates for Interstate Inmate Calling Services

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2013-12-27

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Designing bovine T-cell vaccines via reverse immunology

USDA-ARS?s Scientific Manuscript database

T-cell responses contribute to immunity against many intra-cellular infections. There is, for example, strong evidence that major histocompatibility complex (MHC) class I restricted cytotoxic T lymphocytes (CTLs) play an essential role in mediating immunity to East Coast fever (ECF), a fatal lymphop...

78 FR 1188 - Commercial Operations in the 3550-3650 MHz Band

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2013-01-08

... Operations in the 3550-3650 MHz Band AGENCY: Federal Communications Commission. ACTION: Proposed rule... the following methods: Federal Communications Commission's Web Site: http://fjallfoss.fcc.gov/ecfs2... Federal Communications Commission (Commission) propose to create a new Citizens Broadband Service in the...

Volume versus wiring transmission in the brain: a new theoretical frame for neuropsychopharmacology.

PubMed

Agnati, L F; Bjelke, B; Fuxe, K

1995-01-01

A volume transmission mode of communication in brain was implicit already in the early work of Golgi, who postulated the existence of electrical signals in the extracellular fluid (ECF) based on Volta's "wet conductor" made by solutions. The term volume transmission is taken from the term volume conduction describing the flow of ionic currents in the ECF as a basis for the electrocorticogram. The slow VT mode includes also chemical signals and is opposed to the fast synaptic (wiring) transmission. Every neuron may function in a dual mode, the synaptic and the volume transmission mode, when considering the autocrine and synaptic classes of communication. The paracrine- and neuroendocrine-like classes only involve the VT mode in the latter case including the CSF as a route. The chemical signals for VT are the neuropeptides, but also the classical transmitters, the monoamines, acetylcholine, GABA, and glutamate can participate, when they operate via slow, high affinity G protein coupled receptors. Ions such as K+, Ca++, and H+ also function as VT signals. The hypothesis is also introduced that CO2 can act as a multifacit long-distance VT and WT regulator besides being part of the CO2/HCO3 buffer. CO2 via regulating NMDA receptor sensitivity can also regulate NO formation, which represents a paracrine and fast VT signal. The therapy of CNS disorders is also discussed in the frame of the wiring and VT concept. Two therapeutical approaches can therefore be developed, one based on increasing WT and one based on increasing VT. In contrast to the WT therapy, which must preserve the electrotemporal code, the VT therapy can operate also with postsynaptic agonists. Therefore, a therapeutic effect with such a drug indicates that the deficiency in the communication process operates via VT. In view of the lack of very effective negative feedbacks in VT vs. WT, VT therapy may produce less tolerance and drug dependency.

East Coast fever as a continued constraint to livestock improvement in Tanzania: a case study.

PubMed

Kambarage, D M

1995-08-01

An investigation was carried out into the cause of deaths in a recently established dairy farm with 211 animals. Clinical examination revealed that 14 out of 15 sick animals were depressed, pyrexic, anorexic and had variable degrees of respiratory distress and enlarged lymph nodes. These clinical signs were suggestive of East Coast fever (ECF). This was confirmed on positive demonstration of piroplasms and macroschizonts in blood and lymph node smears respectively, and on post-mortem examination. Parasites were also demonstrated in smears taken from 5 other animals which were pyrexic and had enlarged lymph nodes. Epidemiological investigation revealed that the occurrence of the disease was associated with contact with tick-infested pastures and unsatisfactory tick control due to improper dipping of the herd. The use of pasture by pastoral cattle which are rarely dipped also increased tick infestation. It is concluded that, unless effective disease control is applied ECF will continue to be a major killer disease of cattle in Tanzania.

Second Announcement - ESO/ST-ECF Workshop on NICMOS and the VLT: A New Era of High-Resolution Near-Infrared Imaging and Spectroscopy - May 26-27, 1998 - Hotel Baia di Nora, Pula, Sardinia, Italy

NASA Astrophysics Data System (ADS)

1998-03-01

ST-ECF and ESO are organising in collaboration with the NICMOS IDT and STScI a workshop on near infrared imaging from space and ground. The purpose of the workshop is to review what has been achieved with the Near Infrared and Multi Object Spectrograph (NICMOS) on board of HST, what can be achieved in the remaining lifetime of the instrument, and how NICMOS observations can be optimised taking into account the availability of IR imaging and spectroscopy on ESO's Very large Telescope (VLT) in the near future. The meeting will be held in May 1998, about one year after science observations started with NICMOS, and about half a year before the Infrared Spectrometer and Array Camera (ISAAC) starts to operate on the VLT. Currently, it is expected that NICMOS will operate until the end of 1998.

Low-latitude variability of ice cloud properties and cloud thermodynamic phase observed by the Atmospheric Infrared Sounder (AIRS)

NASA Astrophysics Data System (ADS)

Kahn, B. H.; Yue, Q.; Davis, S. M.; Fetzer, E. J.; Schreier, M. M.; Tian, B.; Wong, S.

2016-12-01

We will quantify the time and space dependence of ice cloud effective radius (CER), optical thickness (COT), cloud top temperature (CTT), effective cloud fraction (ECF), and cloud thermodynamic phase (ice, liquid, or unknown) with the Version 6 Atmospheric Infrared Sounder (AIRS) satellite observational data set from September 2002 until present. We show that cloud frequency, CTT, COT, and ECF have substantially different responses to ENSO variations. Large-scale changes in ice CER are also observed with a several micron tropics-wide increase during the 2015-2016 El Niño and similar decreases during the La Niña phase. We show that the ice CER variations reflect fundamental changes in the spatial distributions and relative frequencies of different ice cloud types. Lastly, the high spatial and temporal resolution variability of the cloud fields are explored and we show that these data capture a multitude of convectively coupled tropical waves such as Kelvin, westward and eastward intertio-gravity, equatorial Rossby, and mixed Rossby-gravity waves.

Three Good Reasons for Celebrating at the ESO/ST-ECF Science Archive Facility

NASA Astrophysics Data System (ADS)

2000-12-01

Great Demand for Data from New "Virtual Observatory" Summary Due to a happy coincidence, the ESO/ST-ECF Science Archive Facility is celebrating three different milestones at the same time: * its 10th anniversary since the establishment in 1991 * the 10,000th request for data , and * the signing-up of active user number 2000 . This Archive contains over 8 Terabytes (1 Terabyte = 1 million million bytes) of valuable observational data from the NASA/ESA Hubble Space Telescope (HST), the ESO Very Large Telescope (VLT) and other ESO telescopes . Its success paves the way for the establishment of "Virtual Observatories" from which first-class data can be obtained by astronomers all over the world. This greatly enhances the opportunities for more (young) scientists to participate in front-line research. PR Photo 34/00 : Front-page of a new brochure, describing the ESO/ST-ECF Science Archive Facility. Just 10 years ago, on the 1st of January 1991, the ESO/ST-ECF (European Southern Observatory/Space Telescope-European Coordinating Facility) Science Archive Facility opened. It has since served the astronomical community with gigabyte after gigabyte of high-quality astronomical data from some of the world's leading telescopes. The Archive, which is located in Garching, just outside Munich (Germany), contains data from the 2.4-m NASA/ESA Hubble Space Telescope , as well as from several ESO telescopes: the four 8.2-m Unit Telescopes of the Very Large Telescope (VLT) at the Paranal Observatory , and the 3.5-m New Technology Telescope (NTT) , the 3.6-m telescope and the MPG/ESO 2.2-m telescope at La Silla. The Archive is a continuously developing project - in terms of amounts of data stored, the number of users and in particular because of the current dramatic development of innovative techniques for data handling and storage. In the year 2000 more than 2 Terabytes (2000 Gigabytes) of data were distributed to users worldwide. The archiving of VLT data has been described in ESO PR 10/99. Celebrating the 10th anniversary Due to a happy coincidence, the Archive passes two other milestones almost exactly at the time of its ten-year anniversary: the 10,000th request for data has just arrived, and active user number 2000 has just signed up to start using the Archive . Dataset number 10000 was requested by Danish astronomer Søren Larsen who works at the University of California (USA). He asked for images of galaxies taken with the Hubble Space Telescope and expressed great satisfaction with the material: "The extremely sharp images from Hubble have provided a quantum leap forward in our ability to study star clusters in external galaxies. We now know that some galaxies contain extremely bright young star clusters. These might constitute a "link" between open and globular clusters as we know them in the Milky Way galaxy in which we live. We are now trying to understand whether all these clusters really form in the same basic way." Active user number 2000 is Swiss astronomer Frédéric Pont , working at the Universidad de Chile: "We use observations from the ESO VLT Unit Telescopes to map the chemical and star-formation history of dwarf galaxies in the Local Group. The stars we are looking at are very faint and we simply need the large size and excellent quality of VLT to observe them in detail. With the new data, we can really move forward in this fundamental research field." ESO PR Photo 34/00 ESO PR Photo 34/00 [Preview - JPEG: 400 x 281 pix - 63k] [Normal - JPEG: 800 x 562 pix - 224k] [Full-Res - JPEG: 1024 x 714 pix - 336k] Caption : PR Photo 34/00 shows the frontpage of the new brochure that describes the ESO/ST-ECF Science Archive Facility (available in PDF version on the web). The collage shows the Hubble Space Telescope above the world's largest optical/infrared telescope, the Very Large Telescope (VLT). To celebrate this special occasion, a 4-page brochure has been prepared that describes the Archive and its various services. The brochure can be requested from ESO or ST-ECF and is now available in PDF format on the web. As a small token, the two astronomers will receive a commemorative version of the photo that accompanies this release. The ASTROVIRTEL initiative One of the major new initiatives undertaken by ESO and ST-ECF in connection with the ESO/ST-ECF Science Archive is ASTROVIRTEL (Accessing Astronomical Archives as Virtual Telescopes) , cf. ESO PR 09/00. It is a project aimed at helping scientists to cope efficiently with the massive amounts of data now becoming available from the world's leading telescopes and so to exploit the true potential of the Archive treasures. ASTROVIRTEL represents the European effort in an area that many astronomers considers one of the most important developments within observing astronomy in the past decade. The future The head of the ESO/ST-ECF Science Archive Facility , Benoît Pirenne , believes that the future holds exciting challenges: "Due to the many improvements of the ESO, NASA and ESA telescopes and instruments expected in the coming years, we anticipate a tremendous increase in the amount of data to be archived and re-distributed. It will not be too long before we will have to start counting storage space in Petabytes (1 Petabyte = 1,000 Terabytes). We are now trying to figure out how to best prepare for this new era." But he is also concerned with maintaining and further enhancing the astronomical value of the data that are made available to the users: "Apart from improving the data storage, we need to invest much effort in building automatic software that will help users with the tedious pre-processing and 'cleaning' of the data, thereby allowing them to focus more on scientific than technical problems."

Influence of biophase distribution and P-glycoprotein interaction on pharmacokinetic-pharmacodynamic modelling of the effects of morphine on the EEG

PubMed Central

Groenendaal, D; Freijer, J; de Mik, D; Bouw, M R; Danhof, M; de Lange, E C M

2007-01-01

Background and purpose: The aim was to investigate the influence of biophase distribution including P-glycoprotein (Pgp) function on the pharmacokinetic-pharmacodynamic correlations of morphine's actions in rat brain. Experimental approach: Male rats received a 10-min infusion of morphine as 4 mg kg−1, combined with a continuous infusion of the Pgp inhibitor GF120918 or vehicle, 10 or 40 mg kg−1. EEG signals were recorded continuously and blood samples were collected. Key results: Profound hysteresis was observed between morphine blood concentrations and effects on the EEG. Only the termination of the EEG effect was influenced by GF120918. Biophase distribution was best described with an extended catenary biophase distribution model, with a sequential transfer and effect compartment. The rate constant for transport through the transfer compartment (k1e) was 0.038 min−1, being unaffected by GF120918. In contrast, the rate constant for the loss from the effect compartment (keo) decreased 60% after GF120918. The EEG effect was directly related to concentrations in the effect compartment using the sigmoidal Emax model. The values of the pharmacodynamic parameters E0, Emax, EC50 and Hill factor were 45.0 μV, 44.5 μV, 451 ng ml−1 and 2.3, respectively. Conclusions and implications: The effects of GF120918 on the distribution kinetics of morphine in the effect compartment were consistent with the distribution in brain extracellular fluid (ECF) as estimated by intracerebral microdialysis. However, the time-course of morphine concentrations at the site of action in the brain, as deduced from the biophase model, is distinctly different from the brain ECF concentrations. PMID:17471181

Breast Cancer-Related Arm Lymphedema: Fluctuation over Six Months and the Effect of the Weather.

PubMed

Czerniec, Sharon A; Ward, Leigh C; Kilbreath, Sharon L

2016-09-01

An understanding of normal fluctuation of lymphedema over time is important to identify real change, whether it is from response to treatment or worsening of the condition. The weather is another factor that possibly influences lymphedema but has had minimal investigation to date. The purpose of this study was to determine the extent to which breast cancer-related lymphedema (BCRL) fluctuated over a 6-month period and the influence of temperature, humidity, and barometric pressure. Women with unilateral BCRL (n = 26) and without BCRL (n = 17) were measured on nine occasions over 6 months using a standardized protocol. Measures included self-reported arm swelling, arm volume, and extracellular fluid with bioimpedance. Daily weather data were obtained for analysis of effects on lymphedema. Neither arm volume nor extracellular fluid varied significantly for women with lymphedema; coefficients of variation were 2.3% and 3.7%, respectively. Women without lymphedema had even less fluctuation, with coefficient of variation of 1.9% for arm volume and 2.9% for ECF. Correlation of weather and lymphedema data showed that temperature was the only aspect of the weather to have any effect on BCRL, with the maximum temperature on the day before measurement slightly affecting ECF (r = 0.27, p < 0.001), arm volume (r = 0.23, p < 0.001), and self-reported swelling (r = 0.26, p < 0.001). For women without lymphedema, the weather did not affect any measure. Established BCRL is relatively stable over a 6-month period. Temperature was the only aspect of the weather found to impact lymphedema.

77 FR 60883 - Special Conditions: Eurocopter France (ECF) Model EC225LP Helicopter, Installation of a Search...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-10-05

...; Sec. 29.923(a)(2), Rotor drive system and control mechanism tests. In addition to the applicable... State: Wave height of 2.5 meters (8.2 feet), considering both short and long swells. (ii) Wind: 25 knots...

78 FR 50415 - Wireline Competition Bureau Seeks Comment on Proposed Sample Reseller Certification Language for...

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2013-08-19

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78 FR 14957 - Wireline Competition Bureau Releases Further Discussion Topics and Seeks Additional Comment in...

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East coast fever caused by Theileria parva is characterized by macrophage activation associated with vasculitis and respiratory failure

USDA-ARS?s Scientific Manuscript database

Respiratory failure and death in East Coast Fever (ECF), a clinical syndrome of African cattle caused by the apicomplexan parasite Theileria parva, has historically been attributed to pulmonary infiltration by infected lymphocytes. However, immunohistochemical staining of tissue from T. parva infect...

76 FR 79607 - Local Number Portability Porting Interval and Validation Requirements; Telephone Number Portability

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2011-12-22

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47 CFR 27.20 - Digital television transition education reports.

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2013-10-01

... filed electronically using the Commission's Electronic Comment File System (ECFS), the “Document Type...—the licensee holding such authorization must file a report with the Commission indicating whether, in... April-June quarter of 2008, the licensee must file its first report by July 10, 2008. Each quarterly...

47 CFR 27.20 - Digital television transition education reports.

Code of Federal Regulations, 2012 CFR

2012-10-01

... filed electronically using the Commission's Electronic Comment File System (ECFS), the “Document Type...—the licensee holding such authorization must file a report with the Commission indicating whether, in... April-June quarter of 2008, the licensee must file its first report by July 10, 2008. Each quarterly...

Regulatory Response to Carbon Starvation in Caulobacter crescentus

PubMed Central

Britos, Leticia; Abeliuk, Eduardo; Taverner, Thomas; Lipton, Mary; McAdams, Harley; Shapiro, Lucy

2011-01-01

Bacteria adapt to shifts from rapid to slow growth, and have developed strategies for long-term survival during prolonged starvation and stress conditions. We report the regulatory response of C. crescentus to carbon starvation, based on combined high-throughput proteome and transcriptome analyses. Our results identify cell cycle changes in gene expression in response to carbon starvation that involve the prominent role of the FixK FNR/CAP family transcription factor and the CtrA cell cycle regulator. Notably, the SigT ECF sigma factor mediates the carbon starvation-induced degradation of CtrA, while activating a core set of general starvation-stress genes that respond to carbon starvation, osmotic stress, and exposure to heavy metals. Comparison of the response of swarmer cells and stalked cells to carbon starvation revealed four groups of genes that exhibit different expression profiles. Also, cell pole morphogenesis and initiation of chromosome replication normally occurring at the swarmer-to-stalked cell transition are uncoupled in carbon-starved cells. PMID:21494595

Conserved ABC Transport System Regulated by the General Stress Response Pathways of Alpha- and Gammaproteobacteria.

PubMed

Herrou, Julien; Willett, Jonathan W; Czyż, Daniel M; Babnigg, Gyorgy; Kim, Youngchang; Crosson, Sean

2017-03-01

Brucella abortus σ E1 is an EcfG family sigma factor that regulates the transcription of dozens of genes in response to diverse stress conditions and is required for maintenance of chronic infection in a mouse model. A putative ATP-binding cassette transporter operon, bab1_0223-bab1_0226 , is among the most highly activated gene sets in the σ E1 regulon. The proteins encoded by the operon resemble quaternary ammonium-compatible solute importers but are most similar in sequence to the broadly conserved YehZYXW system, which remains largely uncharacterized. Transcription of yehZYXW is activated by the general stress sigma factor σ S in Enterobacteriaceae , which suggests a functional role for this transport system in bacterial stress response across the classes Alphaproteobacteria and Gammaproteobacteria We present evidence that B. abortus YehZYXW does not function as an importer of known compatible solutes under physiological conditions and does not contribute to the virulence defect of a σ E1 -null strain. The sole in vitro phenotype associated with genetic disruption of this putative transport system is reduced growth in the presence of high Li + ion concentrations. A crystal structure of B. abortus YehZ revealed a class II periplasmic binding protein fold with significant structural homology to Archaeoglobus fulgidus ProX, which binds glycine betaine. However, the structure of the YehZ ligand-binding pocket is incompatible with high-affinity binding to glycine betaine. This is consistent with weak measured binding of YehZ to glycine betaine and related compatible solutes. We conclude that YehZYXW is a conserved, stress-regulated transport system that is phylogenetically and functionally distinct from quaternary ammonium-compatible solute importers. IMPORTANCE Brucella abortus σ E1 regulates transcription in response to stressors encountered in its mammalian host and is necessary for maintenance of chronic infection in a mouse model. The functions of the majority of genes regulated by σ E1 remain undefined. We present a functional/structural analysis of a conserved putative membrane transport system (YehZYXW) whose expression is strongly activated by σ E1 Though annotated as a quaternary ammonium osmolyte uptake system, experimental physiological studies and measured ligand-binding properties of the periplasmic binding protein (PBP), YehZ, are inconsistent with this function. A crystal structure of B. abortus YehZ provides molecular insight into differences between bona fide quaternary ammonium osmolyte importers and YehZ-related proteins, which form a distinct phylogenetic and functional group of PBPs. Copyright © 2017 American Society for Microbiology.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Herrou, Julien; Willett, Jonathan W.; Czyż, Daniel M.

ABSTRACT Brucella abortusσ E1is an EcfG family sigma factor that regulates the transcription of dozens of genes in response to diverse stress conditions and is required for maintenance of chronic infection in a mouse model. A putative ATP-binding cassette transporter operon,bab1_0223-bab1_0226, is among the most highly activated gene sets in the σ E1regulon. The proteins encoded by the operon resemble quaternary ammonium-compatible solute importers but are most similar in sequence to the broadly conserved YehZYXW system, which remains largely uncharacterized. Transcription ofyehZYXWis activated by the general stress sigma factor σ SinEnterobacteriaceae, which suggests a functional role for this transport systemmore » in bacterial stress response across the classesAlphaproteobacteriaandGammaproteobacteria. We present evidence thatB. abortusYehZYXW does not function as an importer of known compatible solutes under physiological conditions and does not contribute to the virulence defect of a σ E1-null strain. The solein vitrophenotype associated with genetic disruption of this putative transport system is reduced growth in the presence of high Li +ion concentrations. A crystal structure ofB. abortusYehZ revealed a class II periplasmic binding protein fold with significant structural homology toArchaeoglobus fulgidusProX, which binds glycine betaine. However, the structure of the YehZ ligand-binding pocket is incompatible with high-affinity binding to glycine betaine. This is consistent with weak measured binding of YehZ to glycine betaine and related compatible solutes. We conclude that YehZYXW is a conserved, stress-regulated transport system that is phylogenetically and functionally distinct from quaternary ammonium-compatible solute importers. IMPORTANCEBrucella abortusσ E1regulates transcription in response to stressors encountered in its mammalian host and is necessary for maintenance of chronic infection in a mouse model. The functions of the majority of genes regulated by σ E1remain undefined. We present a functional/structural analysis of a conserved putative membrane transport system (YehZYXW) whose expression is strongly activated by σ E1. Though annotated as a quaternary ammonium osmolyte uptake system, experimental physiological studies and measured ligand-binding properties of the periplasmic binding protein (PBP), YehZ, are inconsistent with this function. A crystal structure ofB. abortusYehZ provides molecular insight into differences between bona fide quaternary ammonium osmolyte importers and YehZ-related proteins, which form a distinct phylogenetic and functional group of PBPs.« less

Conserved ABC Transport System Regulated by the General Stress Response Pathways of Alpha- and Gammaproteobacteria

DOE Office of Scientific and Technical Information (OSTI.GOV)

Herrou, Julien; Willett, Jonathan W.; Czyż, Daniel M.

ABSTRACT Brucella abortusσ E1is an EcfG family sigma factor that regulates the transcription of dozens of genes in response to diverse stress conditions and is required for maintenance of chronic infection in a mouse model. A putative ATP-binding cassette transporter operon,bab1_0223-bab1_0226, is among the most highly activated gene sets in the σ E1regulon. The proteins encoded by the operon resemble quaternary ammonium-compatible solute importers but are most similar in sequence to the broadly conserved YehZYXW system, which remains largely uncharacterized. Transcription ofyehZYXWis activated by the general stress sigma factor σ SinEnterobacteriaceae, which suggests a functional role for this transport systemmore » in bacterial stress response across the classesAlphaproteobacteriaandGammaproteobacteria. We present evidence thatB. abortusYehZYXW does not function as an importer of known compatible solutes under physiological conditions and does not contribute to the virulence defect of a σ E1-null strain. The solein vitrophenotype associated with genetic disruption of this putative transport system is reduced growth in the presence of high Li +ion concentrations. A crystal structure ofB. abortusYehZ revealed a class II periplasmic binding protein fold with significant structural homology toArchaeoglobus fulgidusProX, which binds glycine betaine. However, the structure of the YehZ ligand-binding pocket is incompatible with high-affinity binding to glycine betaine. This is consistent with weak measured binding of YehZ to glycine betaine and related compatible solutes. We conclude that YehZYXW is a conserved, stress-regulated transport system that is phylogenetically and functionally distinct from quaternary ammonium-compatible solute importers. IMPORTANCEBrucella abortusσ E1regulates transcription in response to stressors encountered in its mammalian host and is necessary for maintenance of chronic infection in a mouse model. The functions of the majority of genes regulated by σ E1remain undefined. We present a functional/structural analysis of a conserved putative membrane transport system (YehZYXW) whose expression is strongly activated by σ E1. Though annotated as a quaternary ammonium osmolyte uptake system, experimental physiological studies and measured ligand-binding properties of the periplasmic binding protein (PBP), YehZ, are inconsistent with this function. A crystal structure ofB. abortusYehZ provides molecular insight into differences between bona fide quaternary ammonium osmolyte importers and YehZ-related proteins, which form a distinct phylogenetic and functional group of PBPs.« less

Quantification of Theileria parva in Rhipicephalus appendiculatus (Acari: Ixodidae) Confirms Differences in Infection Between Selected Tick Strains

USDA-ARS?s Scientific Manuscript database

Theileria parva is the etiologic agent of East Coast fever (ECF), an economically important disease of cattle in sub-Saharan Africa. This protozoan parasite is biologically transmitted by Rhipicephalus appendiculatus. An understanding of the details of the vector-parasite interaction may aid the d...

76 FR 53326 - Airworthiness Directives; Eurocopter France (ECF) Model EC120B Helicopters

Federal Register 2010, 2011, 2012, 2013, 2014

2011-08-26

... also requires modifying the emergency switch electrical wiring and performing tests to ensure correct... the RFM after modifying the emergency switch electrical wiring and performing tests to ensure correct... likely to exist or develop on other helicopters of the same type design. Differences Between This AD and...

75 FR 22510 - Airworthiness Directives; Eurocopter France (ECF) Model EC120B Helicopters

Federal Register 2010, 2011, 2012, 2013, 2014

2010-04-29

...-0410; Directorate Identifier 2010-SW-024-AD; Amendment 39-16265; AD 2010-05-51] RIN 2120-AA64... Register an amendment adopting Airworthiness Directive (AD) 2010-05-51, which was sent previously to all... scoring, paint flaking or left-over identification plate adhesive, the AD requires sanding the area until...

47 CFR 27.20 - Digital television transition education reports.

Code of Federal Regulations, 2014 CFR

2014-10-01

... File System (ECFS), the “Document Type” on the cover sheet should indicate “REPORT.” (c) The reporting... thereafter as specified in paragraph (c) of this section—the licensee holding such authorization must file a... the license authorization is granted during the April-June quarter of 2008, the licensee must file its...

76 FR 22103 - The Consumer and Governmental Affairs Bureau Seeks Comment on “Need for Speed” Information for...

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2011-04-20

... to make informed choices among competing broadband Internet access services. According to the most... speed and performance required for the range of Internet applications consumers intend to use, and how... electronically using the Internet by accessing the Commission's Electronic Comment Filing System (ECFS): http...

Evaluation of various harvesting methods for high-density microalgae, Aurantiochytrium sp. KRS101.

PubMed

Kim, Kyochan; Shin, Heewon; Moon, Myounghoon; Ryu, Byung-Gon; Han, Jong-In; Yang, Ji-Won; Chang, Yong Keun

2015-12-01

Five technologies, coagulation, electro-flotation (EF), electro-coagulation-flotation (ECF), centrifugation, and membrane filtration, were systematically assessed for their adequacy of harvesting Aurantiochytrium sp. KRS101, a heterotrophic microalgal species that has much higher biomass concentration than photoautotrophic species. Coagulation, EF, and ECF were found to have limited efficiency. Centrifugation was overly powerful to susceptible cells like Aurantiochytrium sp. KRS101, inducing cell rupture and consequently biomass loss of over 13%. Membrane filtration, in particular equipped with an anti-fouling turbulence generator, turned out to be best suited: nearly 100% of harvesting efficiency and low water content in harvested biomass were achieved. With rotation rate increased, high permeate fluxes could be attained even with extremely concentrated biomass: e.g., 219.0 and 135.0 L/m(2)/h at 150.0 and 203.0 g/L, respectively. Dynamic filtration appears to be indeed a suitable means especially to obtain highly concentrated biomass that have no need of dewatering and can be directly processed. Copyright © 2015 Elsevier Ltd. All rights reserved.

Denitrification using a monopolar electrocoagulation/flotation (ECF) process.

PubMed

Emamjomeh, Mohammad M; Sivakumar, Muttucumaru

2009-01-01

Nitrate levels are limited due to health concerns in potable water. Nitrate is a common contaminant in water supplies, and especially prevalent in surface water supplies and shallow wells. Nitrate is a stable and highly soluble ion with low potential for precipitation or adsorption. These properties make it difficult to remove using conventional water treatment methods. A laboratory batch electrocoagulation/flotation (ECF) reactor was designed to investigate the effects of different parameters such as electrolysis time, electrolyte pH, initial nitrate concentration, and current rate on the nitrate removal efficiency. The optimum nitrate removal was observed at a pH range of between 9 and 11. It appeared that the nitrate removal rate was 93% when the initial nitrate concentration and electrolysis time respectively were 100 mg L(-1)-NO(3)(-) and 40 min. The results showed a linear relationship between the electrolysis time for total nitrate removal and the initial nitrate concentration. It is concluded that the electrocoagulation technology for denitrification can be an effective preliminary process when the ammonia byproduct must be effectively removed by the treatment facilities.

Brief Report: Randomized, Double-Blind Comparison of Tenofovir Alafenamide (TAF) vs Tenofovir Disoproxil Fumarate (TDF), Each Coformulated With Elvitegravir, Cobicistat, and Emtricitabine (E/C/F) for Initial HIV-1 Treatment: Week 144 Results.

PubMed

Arribas, José R; Thompson, Melanie; Sax, Paul E; Haas, Bernhard; McDonald, Cheryl; Wohl, David A; DeJesus, Edwin; Clarke, Amanda E; Guo, Susan; Wang, Hui; Callebaut, Christian; Plummer, Andrew; Cheng, Andrew; Das, Moupali; McCallister, Scott

2017-06-01

In 2 double-blind phase 3 trials, 1733 antiretroviral-naive adults were randomized to tenofovir alafenamide (TAF) or tenofovir disoproxil fumarate (TDF), each coformulated with elvitegravir/cobicistat/emtricitabine (E/C/F). At 144 weeks, TAF was superior to TDF in virologic efficacy, with 84.2% vs 80.0% having HIV-1 RNA <50 copies/mL (difference 4.2%; 95% confidence interval: 0.6% to 7.8%). TAF had less impact than TDF on bone mineral density and renal biomarkers. No participants on TAF had renal-related discontinuations vs 12 on TDF (P < 0.001), with no cases of proximal tubulopathy for TAF vs 4 for TDF. There were greater increases in lipids with TAF vs TDF, with no difference in the total cholesterol to high-density lipoprotein ratio. For initial HIV therapy, E/C/F/TAF is superior to E/C/F/TDF in efficacy and bone and renal safety.

Sessile serrated adenoma (SSA) vs. traditional serrated adenoma (TSA).

PubMed

Torlakovic, Emina Emilia; Gomez, Jose D; Driman, David K; Parfitt, Jeremy R; Wang, Chang; Benerjee, Tama; Snover, Dale C

2008-01-01

The morphologic distinction between various serrated polyps of the colorectum may be challenging. The distinction between sessile serrated adenoma (SSA) and traditional serrated adenoma (TSA) may be difficult using currently available criteria mostly based on cytologic characteristics. We have evaluated 66 serrated polyps including 29 SSA, 18 TSA, and 19 hyperplastic polyps for overall shape of the polyps, architectural features of individual crypts, the presence of eosinophilic cytoplasm, size and distribution of the proliferation and maturation zones, as well as Ki-67 and CK20 expression. The extent of the expression of CK20 and Ki-67 could not distinguish between the 3 types of serrated polyps, but the distribution of their expression was very helpful and differences were statistically significant. The distribution of Ki-67+ cells was the single most helpful distinguishing feature of the serrated polyp type (P<0.0001, chi test). Hyperplastic polyps had regular, symmetric, and increased Ki-67 expression. SSA had irregular, asymmetric, and highly variable expression of Ki-67. TSA had low Ki-67 expression, which was limited to "ectopic crypts" and admixed tubular adenomalike areas. In serrated polyps, ectopic crypt formation (ECF) defined by the presence of ectopic crypts with their bases not seated adjacent to the muscularis mucosae was nearly exclusive to TSA and was found in all cases, while the presence of cytologic atypia and eosinophilia of the cytoplasm were characteristic, but not limited to TSA. No evidence of ECF, but nevertheless abnormal distribution of proliferation zone was characteristic of SSA, whereas HP had neither. The presence of the ECF defines TSA in a more rigorous fashion than previous diagnostic criteria and also explains the biologic basis of exuberant protuberant growth associated with TSA and the lack of such growth in SSA. Recognition of this phenomenon may also help in exploring the genetic and molecular basis for differences between SSA and TSA, because these architectural abnormalities may well be a reflection of abnormalities in genetically programmed mucosal development.

Evaluation of quantitative PCR for early diagnosis of Pseudomonas aeruginosa infection in cystic fibrosis: a prospective cohort study.

PubMed

Héry-Arnaud, G; Nowak, E; Caillon, J; David, V; Dirou, A; Revert, K; Munck, M-R; Frachon, I; Haloun, A; Horeau-Langlard, D; Le Bihan, J; Danner-Boucher, I; Ramel, S; Pelletier, M-P; Rosec, S; Gouriou, S; Poulhazan, E; Payan, C; Férec, C; Rault, G; Le Gal, G; Le Berre, R

2017-03-01

Early detection of Pseudomonas aeruginosa lung positivity is a key element in cystic fibrosis (CF) management. PCR has increased the accuracy of detection of many microorganisms. Clinical relevance of P. aeruginosa quantitative PCR (qPCR) in this context is unclear. Our aim was to determine P. aeruginosa qPCR sensitivity and specificity, and to assess the possible time saved by qPCR in comparison with standard practice (culture). A multicentre cohort study was conducted over a 3-year period in 96 patients with CF without chronic P. aeruginosa colonization. Sputum samples were collected at each visit. Conventional culture and two-step qPCR (oprL qPCR and gyrB/ecfX qPCR) were performed for 707 samples. The positivity criteria were based on the qPCR results, defined in a previous study as follow: oprL qPCR positivity alone if bacterial density was <730 CFU/mL or oprL qPCR combined with gyrB/ecfX qPCR if bacterial density was ≥730 CFU/mL. During follow up, 36 of the 96 patients with CF were diagnosed on culture as colonized with P. aeruginosa. This two-step qPCR displayed a sensitivity of 94.3% (95% CI 79.7%-98.6%), and a specificity of 86.3% (95% CI 83.4%-88.7%). It enabled P. aeruginosa acquisition to be diagnosed earlier in 20 patients, providing a median detection time gain of 8 months (interquartile range 3.7-17.6) for them. Implementing oprL and gyrB/ecfX qPCR in the management of patients with CF allowed earlier detection of first P. aeruginosa lung positivity than culture alone. Copyright © 2016 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Diadenosine tetraphosphate stimulates atrial ANP release via A(1) receptor: involvement of K(ATP) channel and PKC.

PubMed

Yuan, Kuichang; Cao, Chunhua; Bai, Guang Yi; Kim, Sung Zoo; Kim, Suhn Hee

2007-07-01

Diadenosine polyphosphates (APnAs) are endogenous compounds and exert diverse cardiovascular functions. However, the effects of APnAs on atrial ANP release and contractility have not been studied. In this study, the effects of diadenosine tetraphosphate (AP4A) on atrial ANP release and contractility, and their mechanisms were studied using isolated perfused rat atria. Treatment of atria with AP4A resulted in decreases in atrial contractility and extracellular fluid (ECF) translocation whereas ANP secretion and cAMP levels in perfusate were increased in a dose-dependent manner. These effects of AP4A were attenuated by A(1) receptor antagonist but not by A(2A) or A(3) receptor antagonist. Other purinoceptor antagonists also did not show any effects on AP4A-induced ANF release and contractility. The increment of ANP release and negative inotropy induced by AP4A was similar to those induced by AP3A, AP5A, and AP6A. Protein kinase A inhibitors accentuated AP4A-induced ANP secretion. In contrast, an inhibitor of phospholipase C, protein kinase C or sarcolemma K(ATP) channel completely blocked AP4A-induced ANP secretion. However, an inhibitor of adenylyl cyclase or mitochondria K(ATP) channel had no significant modification of AP4A effects. These results suggest that AP4A regulates atrial inotropy and ANP release mainly through A(1) receptor signaling involving phospholipase C-protein kinase C and sarcolemmal K(ATP) channel and that protein kinase A negatively modulates the effects of AP4A.

75 FR 43801 - Airworthiness Directives; Eurocopter France (ECF) Model EC225LP Helicopters

Federal Register 2010, 2011, 2012, 2013, 2014

2010-07-27

... time. Also, we use inspect rather than check when referring to an action required by a mechanic as... the various levels of government. Therefore, I certify this AD: 1. Is not a ``significant regulatory... compliance time. Also, we use inspect rather than check when referring to an action required by a mechanic as...

75 FR 79312 - Special Conditions: Eurocopter France (ECF) Model EC225LP Helicopter, Installation of a Search...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-12-20

....923(a)(2), Rotor drive system and control mechanism tests. In addition to the applicable airworthiness... stems from the likelihood of encountering hazards such as inconsistent wave heights, floating debris...-state and wind conditions: (i) Sea-State: Wave height of 2.5 meters (8.2 feet), considering both short...

Executive Cognitive Functions and Impulsivity as Correlates of Risk Taking and Problem Behavior in Preadolescents

ERIC Educational Resources Information Center

Romer, Daniel; Betancourt, Laura; Giannetta, Joan M.; Brodsky, Nancy L.; Farah, Martha; Hurt, Hallam

2009-01-01

Initiation of drug use and other risky behavior in preadolescence is associated with poor developmental outcomes. In this research, we examine models that ascribe the trajectory to (a) weak executive cognitive function (ECF), (b) early manifestation of externalizing problems, or (c) heightened levels of trait impulsivity. We test the explanatory…

76 FR 49401 - Universal Service-Intercarrier Compensation Transformation

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2011-08-10

... comments. Federal Communications Commission's Web Site: http://fjallfoss.fcc.gov/ecfs2/ . Follow the... available on the Commission's Web site at http://www.fcc.gov . Pursuant to Sec. Sec. 1.415 and 1.419 of the... 12th Street, SW., Room CY-B402, Washington, DC 20554; Web site: http://www.bcpiweb.com ; phone: 1-800...

78 FR 26777 - Technological Advisory Council Recommendation for Improving Receiver Performance

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2013-05-08

... paper makes note that an interference limits policy approach may not be appropriate in all cases. Are.... SUMMARY: The FCC's Technological Advisory Council (TAC) has been tasked to study the role of receivers in.../ecfs2/ . [ssquf] Paper Filers: Parties who choose to file by paper must file an original and one copy of...

75 FR 33748 - Amateur Radio Use of the Allocation at 5 MHz

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2010-06-15

... envelope power (PEP). 3. The existing amateur radio use of the 60 meter band represents a balancing of... Comment Filing System (ECFS), (2) the Federal Government's eRulemaking Portal, or (3) by filing paper... transmitter output power in modern amateur radio transceivers is 100 W PEP, and that the present 50 W PEP...

Traumatic colon injury in damage control laparotomy-A multicenter trial: Is it safe to do a delayed anastomosis?

PubMed

Tatebe, Leah Carey; Jennings, Andrew; Tatebe, Ken; Handy, Alexandra; Prajapati, Purvi; Smith, Michael; Do, Tai; Ogola, Gerald O; Gandhi, Rajesh R; Duane, Therese M; Luk, Stephen; Petrey, Laura Bruce

2017-04-01

Delayed colonic anastomosis after damage control laparotomy (DCL) is an alternative to colostomies during a single laparotomy (SL) in high-risk patients. However, literature suggests increased colonic leak rates up to 27% with DCL, and various reported risk factors. We evaluated our regional experience to determine if delayed colonic anastomosis was associated with worse outcomes. A multicenter retrospective cohort study was performed across three Level I trauma centers encompassing traumatic colon injuries from January 2006 through June 2014. Patients with rectal injuries or mortality within 24 hours were excluded. Patient and injury characteristics, complications, and interventions were compared between SL and DCL groups. Regional readmission data were utilized to capture complications within 6 months of index trauma. Of 267 patients, 69% had penetrating injuries, 21% underwent DCL, and the mortality rate was 4.9%. Overall, 176 received primary repair (26 in DCL), 90 had resection and anastomosis (28 in DCL), and 26 had a stoma created (10 end colostomies and 2 loop ileostomies in DCL). Thirty-five of 56 DCL patients had definitive colonic repair subsequent to their index operation. DCL patients were more likely to be hypotensive; require more resuscitation; and suffer acute kidney injury, pneumonia, adult respiratory distress syndrome, and death. Five enteric leaks (1.9%) and three enterocutaneous fistulas (ECF, 1.1%) were identified, proportionately distributed between DCL and SL (p = 1.00, p = 0.51). No difference was seen in intraperitoneal abscesses (p = 0.13) or surgical site infections (SSI, p = 0.70) between cohorts. Among SL patients, pancreas injuries portended an increased risk of intraperitoneal abscesses (p = 0.0002), as did liver injuries in DCL patients (p = 0.06). DCL was not associated with increased enteric leaks, ECF, SSI, or intraperitoneal abscesses despite nearly two-thirds having delayed repair. Despite this being a multicenter study, it is underpowered, and a prospective trial would better demonstrate risks of DCL in colon trauma. Therapeutic study, level IV.

75 FR 28478 - Airworthiness Directives; Eurocopter France (ECF) Model AS332L1 and AS332L2 Helicopters

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-21

... fully tilted the seat shoulder harness could become jammed between the seat and bulkhead. This condition, if not corrected, could result in the shoulder harness binding and causing the inertial reel to... seat backrest is fully tilted, there is a risk of the shoulder harness jamming between the seat and...

75 FR 38956 - Airworthiness Directives; Eurocopter France (ECF) Model SA330F, G, and J; and AS332C, L, L1, and...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-07-07

... control of the helicopter. DATES: We must receive comments on this proposed AD by August 6, 2010..., Aviation Safety Engineer, Rotorcraft Directorate, Regulations and Guidance Group, 2601 Meacham Blvd, Fort.... We will post all comments we receive, without change, to http://www.regulations.gov , including any...

75 FR 57659 - Airworthiness Directives Eurocopter France (ECF) Model SA-365N1, AS-365N2, AS 365 N3, EC 155B...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-09-22

... aviation authority of the European Aviation Safety Agency (EASA). The MCAI AD reports the separation and...: DOT/FAA Southwest Region, Gary Roach, ASW-111, Aviation Safety Engineer, Rotorcraft Directorate... Rulemaking Title 49 of the United States Code specifies the FAA's authority to issue rules on aviation safety...

Patterns in Entrepreneurial Competences as the Perceived Learning Outcomes of Entrepreneurship Education: The Case of Estonian HEIs

ERIC Educational Resources Information Center

Mets, Tõnis; Kozlinska, Inna; Raudsaar, Mervi

2017-01-01

The importance of evaluating the outcomes of entrepreneurship education (EE) has been widely acknowledged, but how to approach the evaluation and what models and measures to use are still subjected to academic debate. In this article, the authors present an application of the European Competence Framework (ECF)--the knowledge-skills-attitude triad…

Suppression of abnormal morphology and extracytoplasmic function sigma activity in Bacillus subtilis ugtP mutant cells by expression of heterologous glucolipid synthases from Acholeplasma laidlawii.

PubMed

Matsuoka, Satoshi; Seki, Takahiro; Matsumoto, Kouji; Hara, Hiroshi

2016-12-01

Glucolipids in Bacillus subtilis are synthesized by UgtP processively transferring glucose from UDP-glucose to diacylglycerol. Here we conclude that the abnormal morphology of a ugtP mutant is caused by lack of glucolipids, since the same morphology arises after abolition of glucolipid production by disruption of pgcA and gtaB, which are involved in UDP-glucose synthesis. Conversely, expression of a monoglucosyldiacylglycerol (MGlcDG) produced by 1,2-diacylglycerol 3-glucosyltransferase from Acholeplasma laidlawii (alMGS) almost completely suppressed the ugtP disruptant phenotype. Activation of extracytoplasmic function (ECF) sigmas (SigM, SigV, and SigX) in the ugtP mutant was decreased by alMGS expression, and was suppressed to low levels by MgSO 4 addition. When alMGS and alDGS (A. laidlawii 1,2-diacylglycerol-3-glucose (1-2)-glucosyltransferase producing diglucosyldiacylglycerol (DGlcDG)) were simultaneously expressed, SigX activation was repressed to wild type level. These observations suggest that MGlcDG molecules are required for maintenance of B. subtilis cell shape and regulation of ECF sigmas, and DGlcDG regulates SigX activity.

Performance assessment and optimization of an irreversible nano-scale Stirling engine cycle operating with Maxwell-Boltzmann gas

NASA Astrophysics Data System (ADS)

Ahmadi, Mohammad H.; Ahmadi, Mohammad-Ali; Pourfayaz, Fathollah

2015-09-01

Developing new technologies like nano-technology improves the performance of the energy industries. Consequently, emerging new groups of thermal cycles in nano-scale can revolutionize the energy systems' future. This paper presents a thermo-dynamical study of a nano-scale irreversible Stirling engine cycle with the aim of optimizing the performance of the Stirling engine cycle. In the Stirling engine cycle the working fluid is an Ideal Maxwell-Boltzmann gas. Moreover, two different strategies are proposed for a multi-objective optimization issue, and the outcomes of each strategy are evaluated separately. The first strategy is proposed to maximize the ecological coefficient of performance (ECOP), the dimensionless ecological function (ecf) and the dimensionless thermo-economic objective function ( F . Furthermore, the second strategy is suggested to maximize the thermal efficiency ( η), the dimensionless ecological function (ecf) and the dimensionless thermo-economic objective function ( F). All the strategies in the present work are executed via a multi-objective evolutionary algorithms based on NSGA∥ method. Finally, to achieve the final answer in each strategy, three well-known decision makers are executed. Lastly, deviations of the outcomes gained in each strategy and each decision maker are evaluated separately.

75 FR 4475 - Airworthiness Directives; Eurocopter France Model AS332L1, AS332L2, and EC225LP Helicopters

Federal Register 2010, 2011, 2012, 2013, 2014

2010-01-28

... Airworthiness Directives; Eurocopter France Model AS332L1, AS332L2, and EC225LP Helicopters AGENCY: Federal.... SUMMARY: We are adopting a new airworthiness directive (AD) for the specified Eurocopter France (ECF... France and are approved for operation in the United States. Pursuant to our bilateral [[Page 4476...

Projet de classification de spectres stellaires IUE à basse résolution par système expert.

NASA Astrophysics Data System (ADS)

Imadache, A.

Le project d'étude porte sur l'utilisation de l'intelligence artificielle en vue d'établir une classification de spectres IUE. Pour la réalisation de ce projet, des liens de collaboration ont été établis entre l'Observatoire Astronomique de Strasbourg et l'équipe ST-ECF à l'ESO.

Rapid and simultaneous determination of twenty amino acids in complex biological and food samples by solid-phase microextraction and gas chromatography-mass spectrometry with the aid of experimental design after ethyl chloroformate derivatization.

PubMed

Mudiam, Mohana Krishna Reddy; Ratnasekhar, Ch; Jain, Rajeev; Saxena, Prem Narain; Chauhan, Abhishek; Murthy, R C

2012-10-15

Amino acids play a vital role as intermediates in many important metabolic pathways such as the biosynthesis of nucleotides, vitamins and secondary metabolites. A sensitive and rapid analytical method has been proposed for the first time for the simultaneous determination of twenty amino acids using solid-phase microextraction (SPME). The protein samples were hydrolyzed by 6M HCl under microwave radiation for 120 min. Then the amino acids were derivatized by ethyl chloroformate (ECF) and the ethoxy carbonyl ethyl esters of amino acids formed were extracted using SPME by direct immersion. Finally the extracted analytes on the SPME fiber were desorbed at 260°C and analyzed by gas chromatography-mass spectrometer (GC-MS) in electron ionization mode. Factors which affect the SPME efficiency were screened by Plackett-Burmann design; most significant factors were optimized with response surface methodology. The optimum conditions for SPME are as follows: pH of 1.7, ionic strength of 733 mg, extraction time of 30 min and fiber of divinyl benzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS). The recovery of all the amino acids was found to be in the range of 89.17-100.98%. The limit of detection (LOD) of all derivatized amino acids in urine, hair and soybean was found to be in the range of 0.20-7.52 μg L(-1), 0.21-8.40 μg L(-1) and 0.18-5.62 μg L(-1), respectively. Finally, the proposed technique was successfully applied for the determination of amino acids in complex biological (hair, urine) and food samples (soybean). The method can find wide applications in the routine analysis of amino acids in any biological as well as food samples. Copyright © 2012 Elsevier B.V. All rights reserved.

On Cirrus Cloud Fields Measured by the Atmospheric Infrared Sounder

NASA Technical Reports Server (NTRS)

Kahn, Brian H.; Eldering, Annmarie; Liou, Kuo Nan

2006-01-01

A viewgraph presentation showing trends in clouds measured by the Atmospheric Infrared Sounder (AIRS) is given. The topics include: 1) Trends in clouds measured by AIRS: Are they reasonable? 2) Single and multilayered cloud trends; 3) Retrievals of thin cirrus D(sub e) and tau: Single-layered cloud only; 4) Relationships between ECF, D(sub e), tau, and T(sub CLD); and 5) MODIS vs. AIRS retrievals.

75 FR 20931 - Airworthiness Directives; Eurocopter France (ECF) Model SA-365N1, AS-365N2, AS 365 N3, EC 155B...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-04-22

... Community. The MCAI AD reports the separation and loss of a stainless steel ring (75 millimeter (mm) in... of a stainless steel ring (75 mm in diameter) from a blade sleeve resulting in severe, high-frequency... information specifies checking the blade sleeve for slippage of the stainless steel ring (75 mm in diameter...

Theoretical study on the mechanism of the gas-phase elimination kinetics of alkyl chloroformates

NASA Astrophysics Data System (ADS)

Alcázar, Jackson J.; Marquez, Edgar; Mora, José R.; Cordova-Sintjago, Tania; Chuchani, Gabriel

2016-03-01

The theoretical calculations on the mechanism of the homogeneous and unimolecular gas-phase elimination kinetics of alkyl chloroformates- ethyl chloroformate (ECF), isopropyl chloroformate (ICF), and sec-butyl chloroformate (SCF) - have been carried out by using CBS-QB3 level of theory and density functional theory (DFT) functionals CAM-B3LYP, M06, MPW1PW91, and PBE1PBE with the basis sets 6-311++G(d,p) and 6-311++G(2d,2p). The chlorofomate compounds with alkyl ester Cβ-H bond undergo thermal decomposition producing the corresponding olefin, HCl and CO2. These homogeneous eliminations are proposed to undergo two different types of mechanisms: a concerted process, or via the formation of an unstable intermediate chloroformic acid (ClCOOH), which rapidly decomposes to HCl and CO2 gas. Since both elimination mechanisms may occur through a six-membered cyclic transition state structure, it is difficult to elucidate experimentally which is the most reasonable reaction mechanism. Theoretical calculations show that the stepwise mechanism with the formation of the unstable intermediate chloroformic acid from ECF, ICF, and SCF is favoured over one-step elimination. Reasonable agreements were found between theoretical and experimental values at the CAM-B3LYP/6-311++G(d,p) level.

Metronidazole and hydroxymetronidazole central nervous system distribution: 1. microdialysis assessment of brain extracellular fluid concentrations in patients with acute brain injury.

PubMed

Frasca, Denis; Dahyot-Fizelier, Claire; Adier, Christophe; Mimoz, Olivier; Debaene, Bertrand; Couet, William; Marchand, Sandrine

2014-01-01

The distribution of metronidazole in the central nervous system has only been described based on cerebrospinal fluid data. However, extracellular fluid (ECF) concentrations may better predict its antimicrobial effect and/or side effects. We sought to explore by microdialysis brain ECF metronidazole distribution in patients with acute brain injury. Four brain-injured patients monitored by cerebral microdialysis received 500 mg of metronidazole over 0.5 h every 8 h. Brain dialysates and blood samples were collected at steady state over 8 h. Probe recoveries were evaluated by in vivo retrodialysis in each patient for metronidazole. Metronidazole and OH-metronidazole were assayed by high-pressure liquid chromatography, and a noncompartmental pharmacokinetic analysis was performed. Probe recovery was equal to 78.8% ± 1.3% for metronidazole in patients. Unbound brain metronidazole concentration-time curves were delayed compared to unbound plasma concentration-time curves but with a mean metronidazole unbound brain/plasma AUC0-τ ratio equal to 102% ± 19% (ranging from 87 to 124%). The unbound plasma concentration-time profiles for OH-metronidazole were flat, with mean average steady-state concentrations equal to 4.0 ± 0.7 μg ml(-1). This microdialysis study describes the steady-state brain distribution of metronidazole in patients and confirms its extensive distribution.

Further In-vitro Characterization of an Implantable Biosensor for Ethanol Monitoring in the Brain

PubMed Central

Secchi, Ottavio; Zinellu, Manuel; Spissu, Ylenia; Pirisinu, Marco; Bazzu, Gianfranco; Migheli, Rossana; Desole, Maria Speranza; O′Neill, Robert D.; Serra, Pier Andrea; Rocchitta, Gaia

2013-01-01

Ethyl alcohol may be considered one of the most widespread central nervous system (CNS) depressants in Western countries. Because of its toxicological and neurobiological implications, the detection of ethanol in brain extracellular fluid (ECF) is of great importance. In a previous study, we described the development and characterization of an implantable biosensor successfully used for the real-time detection of ethanol in the brain of freely-moving rats. The implanted biosensor, integrated in a low-cost telemetry system, was demonstrated to be a reliable device for the short-time monitoring of exogenous ethanol in brain ECF. In this paper we describe a further in-vitro characterization of the above-mentioned biosensor in terms of oxygen, pH and temperature dependence in order to complete its validation. With the aim of enhancing ethanol biosensor performance, different enzyme loadings were investigated in terms of apparent ethanol Michaelis-Menten kinetic parameters, viz. IMAX, KM and linear region slope, as well as ascorbic acid interference shielding. The responses of biosensors were studied over a period of 28 days. The overall findings of the present study confirm the original biosensor configuration to be the best of those investigated for in-vivo applications up to one week after implantation. PMID:23881145

Occurrence, fluxes and sources of perfluoroalkyl substances with isomer analysis in the snow of northern China.

PubMed

Shan, Guoqiang; Chen, Xinwei; Zhu, Lingyan

2015-12-15

In this study, perfluoroalkyl substances (PFASs) and the isomers of perfluorooctanoate (PFOA) and perfluorooctanesulfonate (PFOS) were analyzed in fresh snow samples collected from 19 cities in northern China, 2013. The levels of total PFASs in the snow samples were 33.5-229 ng/L, suggesting heavy atmospheric pollution of PFASs in northern China. PFOA (9.08-107 ng/L), PFOS (3.52-54.3 ng/L), perfluoroheptanoate (PFHpA) (3.66-44.8 ng/L), and perfluorohexanoate (PFHxA) (3.21-23.6 ng/L) were predominant with a summed contribution of 82% to the total PFASs. The particulate matters (PMs) associated PFASs contributed 21.5-56.2% to the total PFASs in the snow, suggesting PMs are vital for the transport and deposition of airborne PFASs. Partitioning of PFASs between PM and dissolved phases was dependent on the carbon chain length and end functional groups. Isomer profiles of PFOA and PFOS in the snow were in agreement with the signature of the historical 3M electrochemical fluorination (ECF) products, suggesting that the ECF products were still produced and used in China. Further source analysis showed that the airborne PFASs in urban area were mainly due to direct release rather than degradation of their precursors. Copyright © 2015 Elsevier B.V. All rights reserved.

Study on the Modification of Bleached Eucalyptus Kraft Pulp Using Birch Xylan

Treesearch

Wenjia Han; Chuanshan Zhao; Thomas Elder; Rendang Yang; Dongho Kim; Yunqiao Pu; Jeffery Hsieh; Arthur J. Ragauskas

2012-01-01

In this study, birch xylan was deposited onto elementally chlorine free (ECF) bleached eucalyptus kraft pulp, and the corresponding changes in physical properties were determined. An aqueous 5% birch xylan solution at pH 9 was added to 5 wt% slurry of bleached kraft eucalyptus fibers, with stirring at 70 C for 15 min after which the pH was adjusted to 5–6. The xylan...

Simultaneous Determination of Fluid Shifts during Thermal Stress in a Small Animal Model,

DTIC Science & Technology

1985-09-01

extracellular fluid voitmie (BCF) was measured using a single injection c- inulin , technique, and plasma voilme (PV) was determined by ca.rdio--yreen dye...using tritiated water, extracell1ular fluid volume (ECF) was measured using a single injection C- inulin technique, and plasma volume (PV) was...space. However, inulin (10) has several advantages over the aforementioned because it Is not metabolized, stored, or incorporated by cells or

Immediately Outcomes of Lower-Income Participants in Minnesota's Universal Access Early Childhood Family Education. Early Childhood Family Education (ECFC) Evaluation Series; Changing Times, Changing Families-Phase II.

ERIC Educational Resources Information Center

Mueller, Marsha R.

The Minnesota Early Childhood Family Education (ECFE) program is a voluntary public school family support and education program for parents of children from birth to kindergarten, and is offered in 360 school districts and the four tribal schools. An evaluation was conducted to learn what types of immediate outcomes could be expected for…

Evaluation of the absorption Ångström exponents for traffic and wood burning in the Aethalometer-based source apportionment using radiocarbon measurements of ambient aerosol

NASA Astrophysics Data System (ADS)

Zotter, Peter; Herich, Hanna; Gysel, Martin; El-Haddad, Imad; Zhang, Yanlin; Močnik, Griša; Hüglin, Christoph; Baltensperger, Urs; Szidat, Sönke; Prévôt, André S. H.

2017-03-01

Equivalent black carbon (EBC) measured by a multi-wavelength Aethalometer can be apportioned to traffic and wood burning. The method is based on the differences in the dependence of aerosol absorption on the wavelength of light used to investigate the sample, parameterized by the source-specific absorption Ångström exponent (α). While the spectral dependence (defined as α values) of the traffic-related EBC light absorption is low, wood smoke particles feature enhanced light absorption in the blue and near ultraviolet. Source apportionment results using this methodology are hence strongly dependent on the α values assumed for both types of emissions: traffic αTR, and wood burning αWB. Most studies use a single αTR and αWB pair in the Aethalometer model, derived from previous work. However, an accurate determination of the source specific α values is currently lacking and in some recent publications the applicability of the Aethalometer model was questioned.Here we present an indirect methodology for the determination of αWB and αTR by comparing the source apportionment of EBC using the Aethalometer model with 14C measurements of the EC fraction on 16 to 40 h filter samples from several locations and campaigns across Switzerland during 2005-2012, mainly in winter. The data obtained at eight stations with different source characteristics also enabled the evaluation of the performance and the uncertainties of the Aethalometer model in different environments. The best combination of αTR and αWB (0.9 and 1.68, respectively) was obtained by fitting the Aethalometer model outputs (calculated with the absorption coefficients at 470 and 950 nm) against the fossil fraction of EC (ECF / EC) derived from 14C measurements. Aethalometer and 14C source apportionment results are well correlated (r = 0.81) and the fitting residuals exhibit only a minor positive bias of 1.6 % and an average precision of 9.3 %. This indicates that the Aethalometer model reproduces reasonably well the 14C results for all stations investigated in this study using our best estimate of a single αWB and αTR pair. Combining the EC, 14C, and Aethalometer measurements further allowed assessing the dependence of the mass absorption cross section (MAC) of EBC on its source. Results indicate no significant difference in MAC at 880 nm between EBC originating from traffic or wood-burning emissions. Using ECF / EC as reference and constant a priori selected αTR values, αWB was also calculated for each individual data point. No clear station-to-station or season-to-season differences in αWB were observed, but αTR and αWB values are interdependent. For example, an increase in αTR by 0.1 results in a decrease in αWB by 0.1. The fitting residuals of different αTR and αWB combinations depend on ECF / EC such that a good agreement cannot be obtained over the entire ECF / EC range using other α pairs. Additional combinations of αTR = 0.8, and 1.0 and αWB = 1.8 and 1.6, respectively, are possible but only for ECF / EC between ˜ 40 and 85 %. Applying α values previously used in the literature such as αWB of ˜ 2 or any αWB in combination with αTR = 1.1 to our data set results in large residuals. Therefore we recommend to use the best α combination as obtained here (αTR = 0.9 and αWB = 1.68) in future studies when no or only limited additional information like 14C measurements are available. However, these results were obtained for locations impacted by black carbon (BC) mainly from traffic consisting of a modern car fleet and residential wood combustion with well-constrained combustion efficiencies. For regions of the world with different combustion conditions, additional BC sources, or fuels used, further investigations are needed.

Rhipicephalus appendiculatus ticks transmit Theileria parva from persistently infected cattle in the absence of detectable parasitemia: implications for East Coast fever epidemiology.

PubMed

Olds, Cassandra L; Mason, Kathleen L; Scoles, Glen A

2018-03-02

East Coast fever (ECF) is a devastating disease of cattle and a significant constraint to improvement of livestock production in sub-Saharan Africa. The protozoan parasite causing ECF, Theileria parva, undergoes obligate sexual stage development in its tick vector Rhipicephalus appendiculatus. Tick-borne acquisition and transmission occurs transstadially; larval and nymphal ticks acquire infection while feeding and transmit to cattle when they feed after molting to the next stage. Much of the current knowledge relating to tick-borne acquisition and transmission of T. parva has been derived from studies performed during acute infections where parasitemia is high. In contrast, tick-borne transmission during the low-level persistent infections characteristic of endemic transmission cycles is rarely studied. Cattle were infected with one of two stocks of T. parva (Muguga or Marikebuni). Four months post-infection when parasites were no longer detectable in peripheral blood by PCR, 500 R. appendiculatus nymphs were fed to repletion on each of the cattle. After they molted to the adult stage, 20 or 200 ticks, respectively, were fed on two naïve cattle for each of the parasite stocks. After adult ticks fed to repletion, cattle were tested for T. parva infection by nested PCR and dot blot hybridization. Once they had molted to adults the ticks that had fed as nymphs on Muguga and Marikebuni infected cattle successfully transmitted Theileria parva to all naïve cattle, even though T. parva infection was not detectable by nested PCR on salivary gland genomic DNA of a sample of individual ticks. However, a salivary gland homogenate from a single Marikebuni infected tick was able to infect primary bovine lymphocytes. Infection was detected by nested p104 PCR in 3 of 4 calves and detected in all 4 calves by T. parva 18S nested PCR/dot blot hybridization. We show that R. appendiculatus ticks are able to acquire T. parva parasites from infected cattle even in the absence of detectable parasitemia. Although infection was undetectable in a sample of individual ticks, cumulatively as few as 20 ticks were able to transmit T. parva to naïve cattle. These results have important implications for our understanding of T. parva transmission by R. appendiculatus in ECF endemic regions.

Mitochondrial phylogeography and population structure of the cattle tick Rhipicephalus appendiculatus in the African Great Lakes region.

PubMed

Amzati, Gaston S; Pelle, Roger; Muhigwa, Jean-Berckmans B; Kanduma, Esther G; Djikeng, Appolinaire; Madder, Maxime; Kirschvink, Nathalie; Marcotty, Tanguy

2018-05-31

The ixodid tick Rhipicephalus appendiculatus is the main vector of Theileria parva, wich causes the highly fatal cattle disease East Coast fever (ECF) in sub-Saharan Africa. Rhipicephalus appendiculatus populations differ in their ecology, diapause behaviour and vector competence. Thus, their expansion in new areas may change the genetic structure and consequently affect the vector-pathogen system and disease outcomes. In this study we investigated the genetic distribution of R. appendiculatus across agro-ecological zones (AEZs) in the African Great Lakes region to better understand the epidemiology of ECF and elucidate R. appendiculatus evolutionary history and biogeographical colonization in Africa. Sequencing was performed on two mitochondrial genes (cox1 and 12S rRNA) of 218 ticks collected from cattle across six AEZs along an altitudinal gradient in the Democratic Republic of Congo, Rwanda, Burundi and Tanzania. Phylogenetic relationships between tick populations were determined and evolutionary population dynamics models were assessed by mismach distribution. Population genetic analysis yielded 22 cox1 and 9 12S haplotypes in a total of 209 and 126 nucleotide sequences, respectively. Phylogenetic algorithms grouped these haplotypes for both genes into two major clades (lineages A and B). We observed significant genetic variation segregating the two lineages and low structure among populations with high degree of migration. The observed high gene flow indicates population admixture between AEZs. However, reduced number of migrants was observed between lowlands and highlands. Mismatch analysis detected a signature of rapid demographic and range expansion of lineage A. The star-like pattern of isolated and published haplotypes indicates that the two lineages evolve independently and have been subjected to expansion across Africa. Two sympatric R. appendiculatus lineages occur in the Great Lakes region. Lineage A, the most diverse and ubiquitous, has experienced rapid population growth and range expansion in all AEZs probably through cattle movement, whereas lineage B, the less abundant, has probably established a founder population from recent colonization events and its occurrence decreases with altitude. These two lineages are sympatric in central and eastern Africa and allopatric in southern Africa. The observed colonization pattern may strongly affect the transmission system and may explain ECF endemic instability in the tick distribution fringes.

Aortic-Radial Pulse Wave Velocity Ratio in End-stage Renal Disease Patients: Association with Age, Body Tissue Hydration Status, Renal Failure Etiology and Five Years of Hemodialysis.

PubMed

Bia, Daniel; Valtuille, Rodolfo; Galli, Cintia; Wray, Sandra; Armentano, Ricardo; Zócalo, Yanina; Cabrera-Fischer, Edmundo

2017-03-01

The etiology of the end-stage renal disease (ESRD) and the hydration status may be involved in the arterial stiffening process observed in hemodialyzed patients. The ratio between carotid-femoral and carotid-radial pulse wave velocity (PWV ratio) was recently proposed to characterize the patient-specific stiffening process. to analyze: (1) the PWV-ratio in healthy and hemodialyzed subjects, analyzing potential changes associated to etiologies of the ESRD, (2) the PWV-ratio and hydration status using multiple-frequency bioimpedance and, (3) the effects of hemodialysis on PWV-ratio in a 5-year follow-up. PWV-ratio was evaluated in 151 patients differentiated by the pathology determining their ESRD. Total body fluid (TBF), intra and extra cellular fluid (ICF, ECF) were measured in 65 of these patients using bioelectrical-impedance. The association between arterial, hemodynamic or fluid parameters was analyzed. PWV-ratio was evaluated in a group of patients (n = 25) 5 years later (follow-up study). PWV-ratio increased in the ESRD cohort with respect to the control group (1.03 ± 0.23 vs. 1.31 ± 0.37; p < 0.001). PWV-ratio in the diabetic nephropathy group was higher than in all other etiological groups (1.61 ± 0.33; p < 0.05). PWV-ratio was associated with TBF (r = -0.238; p < 0.05), ICF (r = -0.323; p < 0.01), ECF/ICF (r = 0.400; p < 0.001) and ECF/TBF (r = 0.403; p < 0.001). PWV-ratio calculated in ESRD patients in 2007 increased 5 years later (1.14 ± 0.32 vs. 1.43 ± 0.44; p < 0.005). PWV-ratio increased the most in patients with diabetic nephropathy. PWV ratio was significantly associated with age and body hydration status, but not with the blood pressure. PWV-ratio could be considered a blood pressure-independent parameter, associated with the age and hydration status of the patient.

Induction of humoral immune response to multiple recombinant Rhipicephalus appendiculatus antigens and their effect on tick feeding success and pathogen transmission.

PubMed

Olds, Cassandra L; Mwaura, Stephen; Odongo, David O; Scoles, Glen A; Bishop, Richard; Daubenberger, Claudia

2016-09-02

Rhipicephalus appendiculatus is the primary vector of Theileria parva, the etiological agent of East Coast fever (ECF), a devastating disease of cattle in sub-Saharan Africa. We hypothesized that a vaccine targeting tick proteins that are involved in attachment and feeding might affect feeding success and possibly reduce tick-borne transmission of T. parva. Here we report the evaluation of a multivalent vaccine cocktail of tick antigens for their ability to reduce R. appendiculatus feeding success and possibly reduce tick-transmission of T. parva in a natural host-tick-parasite challenge model. Cattle were inoculated with a multivalent antigen cocktail containing recombinant tick protective antigen subolesin as well as two additional R. appendiculatus saliva antigens: the cement protein TRP64, and three different histamine binding proteins. The cocktail also contained the T. parva sporozoite antigen p67C. The effect of vaccination on the feeding success of nymphal and adult R. appendiculatus ticks was evaluated together with the effect on transmission of T. parva using a tick challenge model. To our knowledge, this is the first evaluation of the anti-tick effects of these antigens in the natural host-tick-parasite combination. In spite of evidence of strong immune responses to all of the antigens in the cocktail, vaccination with this combination of tick and parasite antigens did not appear to effect tick feeding success or reduce transmission of T. parva. The results of this study highlight the importance of early evaluation of anti-tick vaccine candidates in biologically relevant challenge systems using the natural tick-host-parasite combination.

"One-pot" ethyl chloroformate derivatization and liquid-liquid extraction of reduced glutathione in erythrocyte and its quantitative GC-MS analysis.

PubMed

Manca, Alessandra; Alladio, Eugenio; Massarenti, Paola; Puccinelli, M Paola; De Francesco, Antonella; Del Grosso, Erika; Mengozzi, Giulio; Pazzi, Marco; Vincenti, Marco

2017-12-01

A simple "one-pot" derivatization and liquid-liquid extraction (LLE) procedure was developed for GC-MS analysis of reduced glutathione (GSH) analysis in erythrocytes. The metabolite was extracted by 5% (w/v) TCA, the supernatant treated with ECF and ethanol-pyridine media, the derivative separated and detected by gas chromatography-mass spectrometry using a short non-polar capillary GC column at a high column-head pressure. Total analysis time was 11min. The process was optimized by a Design of Experiment. The method was validated showing a good linearity over the 25.4-813.4μM concentration range, providing satisfactory results in terms of intra-day and inter-day precision as well as an optimal accuracy. The new method was evaluated in a pilot study involving patients with severe protein malnutrition. Comparison of this group with a group of healthy subjects revealed significantly lower GSH concentrations in erythrocytes in the former, thus proving that the described GC-MS method could be employed for fast and simple GSH analysis in clinical studies. Copyright © 2017 Elsevier B.V. All rights reserved.

Isolation of Environmental Bacteria from Surface and Drinking Water in Mafikeng, South Africa, and Characterization Using Their Antibiotic Resistance Profiles

PubMed Central

Mulamattathil, Suma George; Mbewe, Moses; Ateba, Collins Njie

2014-01-01

The aim of this study was to isolate and identify environmental bacteria from various raw water sources as well as the drinking water distributions system in Mafikeng, South Africa, and to determine their antibiotic resistance profiles. Water samples from five different sites (raw and drinking water) were analysed for the presence of faecal indicator bacteria as well as Aeromonas and Pseudomonas species. Faecal and total coliforms were detected in summer in the treated water samples from the Modimola dam and in the mixed water samples, with Pseudomonas spp. being the most prevalent organism. The most prevalent multiple antibiotic resistance phenotype observed was KF-AP-C-E-OT-K-TM-A. All organisms tested were resistant to erythromycin, trimethoprim, and amoxicillin. All isolates were susceptible to ciprofloxacin and faecal coliforms and Pseudomonas spp. to neomycin and streptomycin. Cluster analysis based on inhibition zone diameter data suggests that the isolates had similar chemical exposure histories. Isolates were identified using gyrB, toxA, ecfX, aerA, and hylH gene fragments and gyrB, ecfX, and hylH fragments were amplified. These results demonstrate that (i) the drinking water from Mafikeng contains various bacterial species and at times faecal and total coliforms. (ii) The various bacteria are resistant to various classes of antibiotics. PMID:25105027

Surgery for post-operative entero-cutaneous fistulas: is bowel resection plus primary anastomosis without stoma a safe option to avoid early recurrence? Report on 20 cases by a single center and systematic review of the literature

PubMed Central

LAURO, A.; CIROCCHI, R.; CAUTERO, N.; DAZZI, A.; PIRONI, D.; DI MATTEO, F.M.; SANTORO, A.; FAENZA, S.; PIRONI, L.; PINNA, A.D.

2017-01-01

Background A review was performed on entero-cutaneous fistula (ECF) repair and early recurrence, adding our twenty adult patients (65% had multiple fistulas). Methods The search yielded 4.098 articles but only 15 were relevant: 1.217 patients underwent surgery. The interval time between fistula’s diagnosis and operative repair was between 3 months and 1 year. A bowel resection with primary anastomosis was performed in 1.048 patients, 192 (18.3%) underwent a covering stoma: 856 patients (81.7%) had a fistula takedown in one procedure. Results The patients had 14.3% recurrence and 13.1% mortality rate. In our experience 75% were surgically treated after a period equal or above one year from fistula occurrence: surgery was very demolitive (in 40% remnant small bowel was less than 100 cm). We performed a bowel resection with a hand-sewn anastomosis (95%) without temporary stoma. In-hospital mortality was 0% and at discharge all were back to oral intake with 0% early re-fistulisation. Conclusions Literature supports our experience: ECF takedown could be safely performed after an adequate period of recovery from 3 months to one year from fistula occurrence. In our series primary repair (bowel resection plus reconnection surgery without temporary stoma) avoided an early recurrence without mortality. PMID:29182901

Effect of the Power Balance® band on static balance, hamstring flexibility, and arm strength in adults.

PubMed

Verdan, Princess J R; Marzilli, Thomas S; Barna, Geanina I; Roquemore, Anntionette N; Fenter, Brad A; Blujus, Brittany; Gosselin, Kevin P

2012-08-01

The purpose of this study was to determine the effect of Power Balance® bands on strength, flexibility, and balance. Strength and flexibility were measured using the MicroFit system. Strength was measured via a bicep curl and flexibility via the sit-and-reach method. Balance was measured by the BIODEX System SD. There were 4 different conditions for the balance test: eyes open on a firm surface (EOFS), eyes closed on a firm surface (ECFS), eyes open on a foam surface (EOFoS), and eyes closed on a foam surface (ECFoS). There were 24 subjects in the study (10 men and 14 women). A counterbalance, double-blind, placebo, controlled within-subject design was used. Each of the subjects participated in 3 treatment sessions, consisting of Power Balance®, placebo band, and no band. An alpha level of p ≤ 0.05 was set a priori. There were no significant differences in strength, flexibility, or balance with regard to the treatments used. There was a significant difference between the conditions in the balance test (p = 0.000): EOFS (0.51), ECFS (0.68), EOFoS (0.99), and ECFoS (2.18); however, these were independent of the treatment conditions. The results indicate that the Power Balance® bands did not have an effect on strength, flexibility, or balance.

Metronidazole and Hydroxymetronidazole Central Nervous System Distribution: 1. Microdialysis Assessment of Brain Extracellular Fluid Concentrations in Patients with Acute Brain Injury

PubMed Central

Frasca, Denis; Dahyot-Fizelier, Claire; Adier, Christophe; Mimoz, Olivier; Debaene, Bertrand; Couet, William

2014-01-01

The distribution of metronidazole in the central nervous system has only been described based on cerebrospinal fluid data. However, extracellular fluid (ECF) concentrations may better predict its antimicrobial effect and/or side effects. We sought to explore by microdialysis brain ECF metronidazole distribution in patients with acute brain injury. Four brain-injured patients monitored by cerebral microdialysis received 500 mg of metronidazole over 0.5 h every 8 h. Brain dialysates and blood samples were collected at steady state over 8 h. Probe recoveries were evaluated by in vivo retrodialysis in each patient for metronidazole. Metronidazole and OH-metronidazole were assayed by high-pressure liquid chromatography, and a noncompartmental pharmacokinetic analysis was performed. Probe recovery was equal to 78.8% ± 1.3% for metronidazole in patients. Unbound brain metronidazole concentration-time curves were delayed compared to unbound plasma concentration-time curves but with a mean metronidazole unbound brain/plasma AUC0–τ ratio equal to 102% ± 19% (ranging from 87 to 124%). The unbound plasma concentration-time profiles for OH-metronidazole were flat, with mean average steady-state concentrations equal to 4.0 ± 0.7 μg ml−1. This microdialysis study describes the steady-state brain distribution of metronidazole in patients and confirms its extensive distribution. PMID:24277041

Dogmas and misunderstandings in East Coast fever.

PubMed

Dolan, T T

1999-09-01

East Coast fever (ECF) is the most important tick-borne disease in eastern, central and southern Africa and caused an estimated loss of US $186 million in 1989 in the 11 countries where it occurs. It was brought to southern Africa with cattle from Tanzania in 1901 and, over the next 3 years, devastated the cattle that had survived the rinderpest pandemic of the 1890s. Chemical control of ticks using arsenical compounds was introduced in the early 1900s and became the main control measure for both ticks and the diseases they transmit. This method of control has become less reliable over the last 30 years for many reasons, including reduced government spending on livestock and extension, the cost of acaricides, acaricide resistance, poor management of dips and spray races, and poor application of cattle movement control and quarantine. Significant advances in immunization and treatment have been made in the last 30 years, and more robust integrated strategies combining immunization, reduced frequency of chemical control and treatment are being adopted or considered. Throughout its history, ECF has been a source of great anxiety and cost to farmers, and of intense interest to research workers. Many dogmas and misconceptions have become established, some of which still flourish while others took years to demolish. This paper briefly reviews these as well as the history of the disease and explores recent epidemiological findings and their relevance to applying effective control.

Co-infections determine patterns of mortality in a population exposed to parasite infection.

PubMed

Woolhouse, Mark E J; Thumbi, Samuel M; Jennings, Amy; Chase-Topping, Margo; Callaby, Rebecca; Kiara, Henry; Oosthuizen, Marinda C; Mbole-Kariuki, Mary N; Conradie, Ilana; Handel, Ian G; Poole, E Jane; Njiiri, Evalyne; Collins, Nicola E; Murray, Gemma; Tapio, Miika; Auguet, Olga Tosas; Weir, Willie; Morrison, W Ivan; Kruuk, Loeske E B; Bronsvoort, B Mark de C; Hanotte, Olivier; Coetzer, Koos; Toye, Philip G

2015-03-01

Many individual hosts are infected with multiple parasite species, and this may increase or decrease the pathogenicity of the infections. This phenomenon is termed heterologous reactivity and is potentially an important determinant of both patterns of morbidity and mortality and of the impact of disease control measures at the population level. Using infections with Theileria parva (a tick-borne protozoan, related to Plasmodium) in indigenous African cattle [where it causes East Coast fever (ECF)] as a model system, we obtain the first quantitative estimate of the effects of heterologous reactivity for any parasitic disease. In individual calves, concurrent co-infection with less pathogenic species of Theileria resulted in an 89% reduction in mortality associated with T. parva infection. Across our study population, this corresponds to a net reduction in mortality due to ECF of greater than 40%. Using a mathematical model, we demonstrate that this degree of heterologous protection provides a unifying explanation for apparently disparate epidemiological patterns: variable disease-induced mortality rates, age-mortality profiles, weak correlations between the incidence of infection and disease (known as endemic stability), and poor efficacy of interventions that reduce exposure to multiple parasite species. These findings can be generalized to many other infectious diseases, including human malaria, and illustrate how co-infections can play a key role in determining population-level patterns of morbidity and mortality due to parasite infections.

Plasma-Sprayed Hydroxylapatite-Based Coatings: Chemical, Mechanical, Microstructural, and Biomedical Properties

NASA Astrophysics Data System (ADS)

Heimann, Robert B.

2016-06-01

This contribution discusses salient properties and functions of hydroxylapatite (HA)-based plasma-sprayed coatings, including the effect on biomedical efficacy of coating thickness, phase composition and distribution, amorphicity and crystallinity, porosity and surface roughness, cohesion and adhesion, micro- and nano-structured surface morphology, and residual coating stresses. In addition, it will provide details of the thermal alteration that HA particles undergo in the extremely hot plasma jet that leads to dehydroxylated phases such as oxyhydroxylapatite (OHA) and oxyapatite (OA) as well as thermal decomposition products such as tri-(TCP) and tetracalcium phosphates (TTCP), and quenched phases such as amorphous calcium phosphate (ACP). The contribution will further explain the role of ACP during the in vitro interaction of the as-deposited coatings with simulated body fluid resembling the composition of extracellular fluid (ECF) as well as the in vivo responses of coatings to the ECF and the host tissue, respectively. Finally, it will briefly describe performance profiles required to fulfill biological functions of osteoconductive bioceramic coatings designed to improve osseointegration of hip endoprostheses and dental root implants. In large parts, the content of this contribution is a targeted review of work done by the author and his students and coworkers over the last two decades. In addition, it is considered a stepping stone toward a standard operation procedure aimed at depositing plasma-sprayed bioceramic implant coatings with optimum properties.

Nonlinear Variability of Body Sway in Patients with Phobic Postural Vertigo

PubMed Central

Schniepp, Roman; Wuehr, Max; Pradhan, Cauchy; Novozhilov, Sergej; Krafczyk, Siegbert; Brandt, Thomas; Jahn, Klaus

2013-01-01

Background: Subjective postural imbalance is a key symptom in the somatoform phobic postural vertigo (PPV). It has been assumed that more attentional control of body posture and / or co-contraction of leg muscles during standing is used to minimize the physiological body sway in PPV. Here we analyze nonlinear variability of body sway in patients with PPV in order to disclose changes in postural control strategy associated with PPV. Methods: Twenty patients with PPV and 20 age-matched healthy subjects (HS) were recorded on a stabilometer platform with eyes open (EO), eyes closed (EC), and while standing on a foam rubber with eyes closed (ECF). Spatio-temporal changes of the center of pressure (CoP) displacement were analyzed to assess the structure of postural variability by computing the scaling exponent α and the sample entropy (SEn) of the time series. Results: With EO on firm ground α and SEn of CoP displacement were significantly lower in patients (p < 0.001). For more difficult conditions (EC, ECF) postural variability in PPV assimilated to that of HS. Conclusion: Postural control in PPV patients differs from HS under normal stance condition. It is characterized by a reduced scaling behavior and higher regularity. These changes in the structure of postural variability might suggest an inappropriate attentional involvement with stabilizing strategies, which are used by HS only for more demanding balance tasks. PMID:23966974

Analysis of the Pointing Accuracy of a 6U CubeSat Mission for Proximity Operations and Resident Space Object Imaging

DTIC Science & Technology

2013-05-29

not necessarily express the views of and should not be attributed to ESA. 1 and visual navigation to maneuver autonomously to reduce the size of the...successful orbit and three-dimensional imaging of an RSO, using passive visual -only navigation and real-time near-optimal guidance. The mission design...Kit ( STK ) in the Earth-centered Earth-fixed (ECF) co- ordinate system, loaded to Simulink and transformed to the BFF for calculation of the SRP

Reduction of the closure time of postoperative enterocutaneous fistulas with fibrin sealant.

PubMed

Avalos-González, Jorge; Portilla-deBuen, Eliseo; Leal-Cortés, Caridad Aurea; Orozco-Mosqueda, Abel; Estrada-Aguilar, María del Carmen; Velázquez-Ramírez, Gabriela Abigail; Ambriz-González, Gabriela; Fuentes-Orozco, Clotilde; Guzmán-Gurrola, Aldo Emmerson; González-Ojeda, Alejandro

2010-06-14

To assess whether the use of fibrin sealant shortens the closure time of postoperative enterocutaneous fistulas (ECFs). The prospective case-control study included 70 patients with postoperative ECFs with an output of < 500 mL/d, a fistulous tract of > 2 cm and without any local complication. They were divided into study (n = 23) and control groups (n = 47). Esophageal, gastric and colocutaneous fistulas were monitored under endoscopic visualization, which also allowed fibrin glue application directly through the external hole. Outcome variables included closure time, time to resume oral feeding and morbidity related to nutritional support. There were no differences in mean age, fistula output, and follow-up. Closure-time for all patients of the study group was 12.5 +/- 14.2 d and 32.5 +/- 17.9 d for the control group (P < 0.001), and morbidity related to nutritional support was 8.6% and 42.5%, respectively (P < 0.01). In patients with colonic fistulas, complete closure occurred 23.5 +/- 19.5 d after the first application of fibrin glue, and spontaneous closure was observed after 36.2 +/- 22.8 d in the control group (P = 0.36). Recurrences were observed in 2 patients because of residual disease. One patient of each group died during follow-up as a consequence of septic complications related to parenteral nutrition. Closure time was significantly reduced with the use of fibrin sealant, and oral feeding was resumed faster. We suggest the use of fibrin sealant for the management of stable enterocutaneous fistulas.

Evaluation of buparvaquone (BUTA-Kel KELA, Belgium) as a treatment of East Coast fever in cattle, in the peri-urban of Dar Es Salaam city, Tanzania.

PubMed

Mbwambo, H A; Magwisha, H B; Mfinanga, J M

2006-06-30

Evaluation trials of the efficacy of buparvaquone (BUTA-kel KELA Laboratoria, N.V. Belgium), as a treatment of field cases of Theileria parva infection (East Coast fever - ECF) were carried out on 63 cattle in the peri-urban of Dar Es Salaam city, Tanzania, during the period November 2004 to August 2005. Thirty-two cattle (56%) received single-dose treatment (2.5 mg buparvaquone per kg body weight), while two and three-dose treatment with interval(s) of 48 h was given to 33% and 11% of total treated cattle, respectively; 38 cattle (60.3%) were treated at an early stage of the disease, while 25 cattle (39.7%) were treated at an advanced stage of the disease. The rectal body temperature of 90.5% of buparvaquone-treated cattle dropped to normal values (37.5-39.5 degrees C) by day 7 of treatment, and by day 15 of treatment 96.8% of treated cattle showed normal values. Pulmonary signs were observed in 8/68 (11.8%) of total ECF diagnosed cattle and were successfully treated, albeit with parvaquone plus frusemide (Fruvexon); were not included in final evaluation of the efficacy of BUTA-kel. The present evaluation trials record a recovery rate of 95.2%. Buparvaquone (BUTA-kel KELA Laboratoria, N.V. Belgium), therefore, records another efficacious and valuable alternative treatment against East Coast fever in Tanzania.

The role of ependymin in the development of long lasting synaptic changes.

PubMed

Shashoua, V E

1.) Three types of training experiments (a complex motor task, avoidance conditioning and classical conditioning) in the goldfish and one in the mouse (T-maze learning) indicate that the brain extracellular glycoprotein (ependymin) has a role in the consolidation process of long-term memory formation. 2.) Direct ELISA measures of the concentration of ependymin in the brain extracellular fluid (ECF) indicate that its level decreases after goldfish learn to associate a light stimulus (cs) with the subsequent arrival of a shock (US): paired CS-US gave changes whereas an unpaired presentation of CS-US gave no changes in comparison to unstimulated controls. 3.) Ependymin is released into ECF and CSF as mixtures of three types of disulfide-linked dimers of two acidic polypeptide chains (M. W. 37 kDa and 31 kDa). It contains 10% carbohydrate as an N-linked glycan. 4.) Ependymin has the capacity to polymerize in response to events that deplete Ca2+ from the brain extracellular environment. A molecular hypothesis relating polymerization properties to the process of formation of long-lasting synaptic changes is proposed. 5.) Investigations of the pattern of regeneration of goldfish optic nerve and the mechanisms of long-term potentiation (LTP) of rat brain hippocampal slices suggest that ependymin has a role in the formation of long-lasting synaptic changes. The E.M. data show that polymerized products which stain with anti-ependymin sera accumulate at synapses and in new spines after LTP.

The Impact of a Submaximal Level of Exercise on Balance Performance in Older Persons

PubMed Central

2014-01-01

Objective. The purpose of this study was to determine the impact of a submaximal level of exercise on balance performance under a variety of conditions. Material and Method. Thirteen community-dwelling older persons with intact foot sensation (age = 66.69 ± 8.17 years, BMI = 24.65 ± 4.08 kg/m2, female, n = 6) volunteered to participate. Subjects' balance performances were measured using the Modified Clinical Test of Sensory Integration of Balance (mCTSIB) at baseline and after test, under four conditions of stance: (1) eyes-opened firm-surface (EOF), (2) eyes-closed firm-surface (ECF), (3) eyes-opened soft-surface (EOS), and (4) eyes-closed soft-surface (ECS). The 6-minute walk test (6MWT) protocol was used to induce the submaximal level of exercise. Data was analyzed using the Wilcoxon Signed-Rank Test. Results. Balance changes during EOF (z = 0.00, P = 1.00) and ECF (z = −1.342, P = 0.180) were not significant. However, balance changes during EOS (z = −2.314, P = 0.021) and ECS (z = −3.089, P = 0.02) were significantly dropped after the 6MWT. Conclusion. A submaximal level of exercise may influence sensory integration that in turn affects balance performance, particularly on an unstable surface. Rehabilitation should focus on designing intervention that may improve sensory integration among older individuals with balance deterioration in order to encourage functional activities. PMID:25383386

In vivo monitoring of multiple trace metals in the brain extracellular fluid of anesthetized rats by microdialysis-membrane desalter-ICPMS.

PubMed

Chung, Y T; Ling, Y C; Yang, C S; Sun, Y C; Lee, P L; Lin, C Y; Hong, C C; Yang, M H

2007-12-01

We have developed an on-line analytical system involving microdialysis (MD) sampling, a carbohydrate membrane desalter (CMD), and an inductively coupled plasma mass spectrometer (ICPMS) system for the simultaneous determination of multiple trace metals in the extracellular fluid (ECF) in the brains of anesthetized rats. The microdialysate that perfused from the animal at a flow rate of 0.5 microL/min was on-line transferred to the CMD to remove the high-sodium matrix, followed by ICPMS measurement. The role of the CMD in this on-line system was investigated in detail. With prior addition of EDTA to the microdialysate to form anionic complexes of the metal analytes and the use of NH4Cl as a regenerant to exchange Na(+) with NH4(+) ions, both quantitative recovery of the trace metal analytes and quantitative removal of the sodium matrix could be achieved. Two experimental modes of the monitoring system were constructed. For those metals (e.g., Cu, Zn, and Mn) that existed at (sub)nanogram-per-milliliter concentrations in the microdialysate, the temporal resolution was 10 min when using a 10 microL loop for sample collection, followed by CMD and ICPMS; for those elements (e.g., Ca and Mg) that existed at microgram-per-milliliter levels (or greater), near-real-time analysis was possible because the microdialysate could be led, bypassing the sample loop, directly to the CMD for desalting without any time delay. Further improvement of the temporal resolution for the low-concentration elements was not possible without decreasing the detection limits of mass detection. Among the eight trace metals tested using this on-line system, the method detection limits for Cu, Zn, Mn, Co, Ni, and Pb reached subnanogram-per-milliliter levels; for electrolyte species such as Ca and Mg, the detection limits were in the range of 50-100 ng/mL. Analytical accuracy, expressed as spike recovery, was 100% +/- 15% for all of the elements tested. We demonstrate the applicability of the proposed system through the successful measurement of the basal values of Ca, Mg, Cu, Zn, and Mn in the ECF of a living rat brain and through in vivo monitoring of the concentration profiles of Mn and Pt in the ECF after the injection of drugs (MnCl2 and cisplatin) into the rats. This microdialysis system is the first to offer real-time, in vivo monitoring of trace elements such as Ca and Mg.

The Bacillus subtilis GntR family repressor YtrA responds to cell wall antibiotics.

PubMed

Salzberg, Letal I; Luo, Yun; Hachmann, Anna-Barbara; Mascher, Thorsten; Helmann, John D

2011-10-01

The transglycosylation step of cell wall synthesis is a prime antibiotic target because it is essential and specific to bacteria. Two antibiotics, ramoplanin and moenomycin, target this step by binding to the substrate lipid II and the transglycosylase enzyme, respectively. Here, we compare the ramoplanin and moenomycin stimulons in the Gram-positive model organism Bacillus subtilis. Ramoplanin strongly induces the LiaRS two-component regulatory system, while moenomycin almost exclusively induces genes that are part of the regulon of the extracytoplasmic function (ECF) σ factor σ(M). Ramoplanin additionally induces the ytrABCDEF and ywoBCD operons, which are not part of a previously characterized antibiotic-responsive regulon. Cluster analysis reveals that these two operons are selectively induced by a subset of cell wall antibiotics that inhibit lipid II function or recycling. Repression of both operons requires YtrA, which recognizes an inverted repeat in front of its own operon and in front of ywoB. These results suggest that YtrA is an additional regulator of cell envelope stress responses.

The Bacillus subtilis GntR Family Repressor YtrA Responds to Cell Wall Antibiotics▿§

PubMed Central

Salzberg, Letal I.; Luo, Yun; Hachmann, Anna-Barbara; Mascher, Thorsten; Helmann, John D.

2011-01-01

The transglycosylation step of cell wall synthesis is a prime antibiotic target because it is essential and specific to bacteria. Two antibiotics, ramoplanin and moenomycin, target this step by binding to the substrate lipid II and the transglycosylase enzyme, respectively. Here, we compare the ramoplanin and moenomycin stimulons in the Gram-positive model organism Bacillus subtilis. Ramoplanin strongly induces the LiaRS two-component regulatory system, while moenomycin almost exclusively induces genes that are part of the regulon of the extracytoplasmic function (ECF) σ factor σM. Ramoplanin additionally induces the ytrABCDEF and ywoBCD operons, which are not part of a previously characterized antibiotic-responsive regulon. Cluster analysis reveals that these two operons are selectively induced by a subset of cell wall antibiotics that inhibit lipid II function or recycling. Repression of both operons requires YtrA, which recognizes an inverted repeat in front of its own operon and in front of ywoB. These results suggest that YtrA is an additional regulator of cell envelope stress responses. PMID:21856850

Working Papers in Acquisition of Knowledge for Image Understanding Research,

DTIC Science & Technology

1976-12-15

Rmvt 112,7,P IM Y. K3p ’Of’ z4-" odK zw Mw; OW, ev 6 "ONO T;I 11; "VI lip, IA wo 44 jo . ... . ...... I I WORKING PAPERS IN ACQUISITION OF...paradigm than any other. They have collected protocolt on subjects verbally describing scenes, after examining them visually. The control variable they... collection of Elements of Control Flow (ECF). To avoid creation of either file, the user presses the return key without typing a name to the respective

Temperature- and Pressure-Dependent Kinetics Studies of Atomic Chlorine Reactions with Some Fluorinated Olefins and Ethers

NASA Astrophysics Data System (ADS)

Nicovich, J. M.; Wine, P. H.; Mazumder, S.; Hatzis, G. P.; Jiang, M.

2016-12-01

Laser flash photolysis of Cl2CO/N2/CO2/X mixtures (X = CF3CH=CH2, E-CF3CH=CHCl, E-CF3CH=CHF, (CF3)2CHOCH3, and CF3CF2CH2OCH3), has been coupled with time-resolved detection of Cl atoms by atomic resonance fluorescence spectroscopy to study the kinetics of Cl + X reactions as a function of temperature (T) and pressure (P). The Cl + X reactions were chosen for study because (1) the compounds X are potentially useful substitutes for ozone depleting substances in practical applications, (2) literature results demonstrate that Cl + X reactions are 60-120 times faster than the corresponding OH + X reactions at ambient T and P, suggesting that reaction with Cl is a significant atmospheric loss process for the compounds X, and (3) to our knowledge, Cl + X kinetics have not previously been studied as functions of T and P. For the Cl + olefin reactions, falloff curves are measured over the approximate T ranges 220-300 K. At elevated temperatures, equilibrium constants are determined for Cl + olefin association/dissociation, thus allowing C-Cl bond strengths in the product haloalkyl radicals to be evaluated. Arrhenius expressions are determined for the (P-independent) Cl + ether reactions over the approximate range of T 200-500K. The Cl + CF3CF2CH2OCH3 reaction is quite fast and its rate coefficient is nearly independent of T. The Cl + (CF3)2CHOCH3 reaction is somewhat slower and has a non-Arrhenius T dependence.

Development of a simultaneous PET/microdialysis method to identify the optimal dose of 11C-raclopride for small animal imaging

PubMed Central

Schiffer, Wynne K.; Alexoff, David L.; Shea, Colleen; Logan, Jean; Dewey, Stephen L.

2005-01-01

In the field of small animal positron emission tomography (PET), the assumptions underlying human and primate kinetic models may not be sustained in rodents. That is, the threshold dose at which a pharmacologic response occurs may be lower in small animals. In order to define this relationship, we combined microPET imaging using 11C-raclopride with microdialysis measures of extracellular fluid (ECF) dopamine (DA). In addition, we performed a series of studies in which a known mass of raclopride was microinfused into one striatum prior to a high specific activity (SA) systemic injection of 11C-raclopride. This single-injection approach provided a high and low SA region of radiotracer binding in the same animal during the same scanning session. Our data demonstrate that the binding potential (BP) declines above 3.5 pmol/ml (0.35 μg), with an ED50 of 8.55 ± 5.62 pmol/ml. These data also provide evidence that BP may be compromised by masses of raclopride below 2.0 pmol/ml (0.326 μg). Increases in ECF DA were produced by mass doses of raclopride over 3.9 pmol/ml (0.329 μg) with an ED50 of 8.53 ± 2.48 pmol/ml. Taken together, it appears that an optimal range of raclopride mass exists between 2.0 and 3.5 pmol/ml, around which the measured BP can be compromised by system sensitivity, endogenous DA, or excessive competition with unlabeled compound. PMID:15848236

Trends of selected cattle diseases in eastern Zambia between 1988 and 2008.

PubMed

Mubamba, Chrisborn; Sitali, Joseph; Gummow, Bruce

2011-09-01

Livestock diseases have long been a challenge to livestock production and public health in sub-Saharan Africa and Zambia in particular. The Eastern Province of Zambia is one area in Zambia that is not spared by this challenge. Among various livestock diseases affecting cattle in this region, the most prominent are East Coast Fever (ECF) and African Animal Trypanasomiasis (AAT). Since little has been published on the epidemiological trends of these diseases in eastern Zambia, a retrospective epidemiological study was carried out using reports that were submitted to the provincial veterinary office over the past 20 years. This paper assists in evaluating the impact of some of these aid programmes. Data was analysed using Excel(©), SPSS(®), Epi Info(©), and Epi Map(©) software. Apparent prevalence of AAT in cattle had decreased in the study period from estimates as high as 50% in Katete and Petauke district in 1990 and 1992 respectively to just below 3% (Petauke and Katete) in 2008, thereby, reducing the provincial apparent prevalence from 20% in 1992 to just below 3% in 2008. AAT apparent prevalence dropped from estimates as high as 17% in Chadiza district and 6% in Chipata district in 1990 to just below 1% in 2008 thereby reducing the provincial mean prevalence of East Coast Fever from 6% (1990) to 1% (2008). The inclusion of donor assistance in disease control programmes for both AAT and ECF appeared to have a significant impact on the prevalence of both diseases. Copyright © 2011 Elsevier B.V. All rights reserved.

Identifying osteoporotic vertebral endplate and cortex fractures

PubMed Central

Santiago, Fernando Ruiz; Deng, Min; Nogueira-Barbosa, Marcello H.

2017-01-01

Osteoporosis is the most common metabolic bone disease, and vertebral fractures (VFs) are the most common osteoporotic fracture. A single atraumatic VF may lead to the diagnosis of osteoporosis. Prevalent VFs increase the risk of future vertebral and non-vertebral osteoporotic fracture independent of bone mineral density (BMD). The accurate and clear reporting of VF is essential to ensure patients with osteoporosis receive appropriate treatment. Radiologist has a vital role in the diagnosis of this disease. Several morphometrical and radiological methods for detecting osteoporotic VF have been proposed, but there is no consensus regarding the definition of osteoporotic VF. A vertebra may fracture yet not ever result in measurable changes in radiographic height or area. To overcome these difficulties, algorithm-based qualitative approach (ABQ) was developed with a focus on the identification of change in the vertebral endplate. Evidence of endplate fracture (rather than variation in vertebral shape) is the primary indicator of osteoporotic fracture according to ABQ criteria. Other changes that may mimic osteoporotic fractures should be systemically excluded. It is also possible that vertebral cortex fracture may not initially occur in endplate. Particularly, vertebral cortex fracture can occur in anterior vertebral cortex without gross vertebral deformity (VD), or fractures deform the anterior vertebral cortex without endplate disruption. This article aims to serve as a teaching material for physicians or researchers to identify vertebral endplate/cortex fracture (ECF). Emphasis is particularly dedicated to identifying ECF which may not be associated apparent vertebral body collapse. We believe a combined approach based on standardized radiologic evaluation by experts and morphometry measurement is the most appropriate approach to detect and classify VFs. PMID:29184768

Genetic diversity and population structure of Theileria parva in South Sudan.

PubMed

Salih, Diaeldin A; Mwacharo, Joram M; Pelle, Roger; Njahira, Moses N; Odongo, David O; Mbole-Kariuki, Mary N; Marcellino, Wani L; Malak, Agol K; Kiara, Henary; El Hussein, Abdel Rahim M; Bishop, Richard P; Skilton, Robert A

2018-05-01

Theileria parva is a parasitic protozoan that causes East Coast fever (ECF), an economically important disease of cattle in eastern, central and southern Africa. In South Sudan, ECF is considered a major constraint for livestock development in regions where the disease is endemic. To obtain insights into the dynamics of T. parva in South Sudan, population genetic analysis was performed. Out of the 751 samples included in this study, 178 blood samples were positive for T. parva by species-specific PCR, were collected from cattle from four regions in South Sudan (Bor = 62; Juba = 45; Kajo keji = 41 and Yei = 30) were genotyped using 14 microsatellite markers spanning the four chromosomes. The T. parva Muguga strain was included in the study as a reference. Linkage disequilibrium was evident when populations from the four regions were treated as a single entity, but, when populations were analyzed separately, linkage disequilibrium was observed in Bor, Juba and Kajo keji. Juba region had a higher multiplicity of infection than the other three regions. Principal components analysis revealed a degree of sub-structure between isolates from each region, suggesting that populations are partially distinct, with genetic exchange and gene flow being limited between parasites in the four geographically separated populations studied. Panmixia was observed within individual populations. Overall T. parva population genetic analyses of four populations in South Sudan exhibited a low level of genetic exchange between the populations, but a high level of genetic diversity within each population. Copyright © 2018 Elsevier GmbH. All rights reserved.

Reduction of the closure time of postoperative enterocutaneous fistulas with fibrin sealant

PubMed Central

Avalos-González, Jorge; Portilla-deBuen, Eliseo; Leal-Cortés, Caridad Aurea; Orozco-Mosqueda, Abel; del Carmen Estrada-Aguilar, María; Velázquez-Ramírez, Gabriela Abigail; Ambriz-González, Gabriela; Fuentes-Orozco, Clotilde; Guzmán-Gurrola, Aldo Emmerson; González-Ojeda, Alejandro

2010-01-01

AIM: To assess whether the use of fibrin sealant shortens the closure time of postoperative enterocutaneous fistulas (ECFs). METHODS: The prospective case-control study included 70 patients with postoperative ECFs with an output of < 500 mL/d, a fistulous tract of > 2 cm and without any local complication. They were divided into study (n = 23) and control groups (n = 47). Esophageal, gastric and colocutaneous fistulas were monitored under endoscopic visualization, which also allowed fibrin glue application directly through the external hole. Outcome variables included closure time, time to resume oral feeding and morbidity related to nutritional support. RESULTS: There were no differences in mean age, fistula output, and follow-up. Closure-time for all patients of the study group was 12.5 ± 14.2 d and 32.5 ± 17.9 d for the control group (P < 0.001), and morbidity related to nutritional support was 8.6% and 42.5%, respectively (P < 0.01). In patients with colonic fistulas, complete closure occurred 23.5 ± 19.5 d after the first application of fibrin glue, and spontaneous closure was observed after 36.2 ± 22.8 d in the control group (P = 0.36). Recurrences were observed in 2 patients because of residual disease. One patient of each group died during follow-up as a consequence of septic complications related to parenteral nutrition. CONCLUSION: Closure time was significantly reduced with the use of fibrin sealant, and oral feeding was resumed faster. We suggest the use of fibrin sealant for the management of stable enterocutaneous fistulas. PMID:20533600

Prenatal programming of renal salt wasting resets postnatal salt appetite, which drives food intake in the rat.

PubMed

Alwasel, Saleh H; Barker, David J P; Ashton, Nick

2012-03-01

Sodium retention has been proposed as the cause of hypertension in the LP rat (offspring exposed to a maternal low-protein diet in utero) model of developmental programming because of increased renal NKCC2 (Na+/K+/2Cl- co-transporter 2) expression. However, we have shown that LP rats excrete more rather than less sodium than controls, leading us to hypothesize that LP rats ingest more salt in order to maintain sodium balance. Rats were fed on either a 9% (low) or 18% (control) protein diet during pregnancy; male and female offspring were studied at 4 weeks of age. LP rats of both sexes held in metabolism cages excreted more sodium and urine than controls. When given water to drink, LP rats drank more and ate more food than controls, hence sodium intake matched excretion. However, when given a choice between saline and water to drink, the total volume of fluid ingested by LP rats fell to control levels, but the volume of saline taken was significantly larger [3.8±0.1 compared with 8.8±1.3 ml/24 h per 100 g of body weight in control and LP rats respectively; P<0.001]. Interestingly food intake also fell to control levels. Total body sodium content and ECF (extracellular fluid) volumes were greater in LP rats. These results show that prenatal programming of renal sodium wasting leads to a compensatory increase in salt appetite in LP rats. We speculate that the need to maintain salt homoeostasis following malnutrition in utero stimulates greater food intake, leading to accelerated growth and raised BP (blood pressure).

Real-Time, Single-Step Bioassay Using Nanoplasmonic Resonator With Ultra-High Sensitivity

NASA Technical Reports Server (NTRS)

Zhang, Xiang (Inventor); Chen, Fanqing Frank (Inventor); Su, Kai-Hang (Inventor); Wei, Qi-Huo (Inventor); Ellman, Jonathan A. (Inventor); Sun, Cheng (Inventor)

2014-01-01

A nanoplasmonic resonator (NPR) comprising a metallic nanodisk with alternating shielding layer(s), having a tagged biomolecule conjugated or tethered to the surface of the nanoplasmonic resonator for highly sensitive measurement of enzymatic activity. NPRs enhance Raman signals in a highly reproducible manner, enabling fast detection of protease and enzyme activity, such as Prostate Specific Antigen (paPSA), in real-time, at picomolar sensitivity levels. Experiments on extracellular fluid (ECF) from paPSA-positive cells demonstrate specific detection in a complex bio-fluid background in real-time single-step detection in very small sample volumes.

Real-time, single-step bioassay using nanoplasmonic resonator with ultra-high sensitivity

DOEpatents

Zhang, Xiang; Ellman, Jonathan A; Chen, Fanqing Frank; Su, Kai-Hang; Wei, Qi-Huo; Sun, Cheng

2014-04-01

A nanoplasmonic resonator (NPR) comprising a metallic nanodisk with alternating shielding layer(s), having a tagged biomolecule conjugated or tethered to the surface of the nanoplasmonic resonator for highly sensitive measurement of enzymatic activity. NPRs enhance Raman signals in a highly reproducible manner, enabling fast detection of protease and enzyme activity, such as Prostate Specific Antigen (paPSA), in real-time, at picomolar sensitivity levels. Experiments on extracellular fluid (ECF) from paPSA-positive cells demonstrate specific detection in a complex bio-fluid background in real-time single-step detection in very small sample volumes.

Detoxification of kraft pulp ECF bleaching effluents by catalytic hydrotreatment.

PubMed

Calvo, L; Gilarranz, M A; Casas, J A; Mohedano, A F; Rodríguez, J J

2007-02-01

Two different effluents from the D(1) and E(1) stages of the ECF bleaching of Eucalyptus globulus kraft pulp were treated by catalytic hydrogenation in a trickle bed reactor using commercial and homemade Pd/AC catalysts. The reactor was fed with the bleaching effluent and a H(2)/N(2) gas stream. The variables studied were space-time (1.4-5g(cat)min/mL), gas to liquid flow ratio (286-1000vol.), gas feed concentration (H(2):N(2), 1:1-1:7.3vol.), temperature (25-100 degrees C) and pressure (1-11bar). Hydrotreatment performance was evaluated in terms of ecotoxicity, adsorbable organic halogen (AOX), chemical oxygen demand (COD), biological oxygen demand (BOD(5)) and colour removal. In all the runs, the ecotoxicity of the effluents decreased as a result of the treatment, achieving reductions that ranged from 70% to 98%. Simultaneously to the reduction of toxicity, the hydrotreatment led to a decrease of the colour of the effluents, being the decrease significantly higher in the case of E(1) effluent. The AOX content was reduced by 85% and 23% for E(1) and D(1) effluents, respectively. In the case of D(1) effluent the removal of ecotoxicity was significantly higher than that of AOX, which indicates that much of the toxicity of the effluent must be associated to non-chlorinated organics. In spite of the important reduction of ecotoxicity, the biodegradability of the effluents only increased slightly. The homemade catalysts, prepared from activated carbons with a high external or non-microporous surface area and mesopore volume and a convenient surface chemistry showed a higher efficiency than the commercial one.

Evaluation of real time PCR assays for the detection and enumeration of enterohemorrhagic Escherichia coli directly from cattle feces.

PubMed

Luedtke, Brandon E; Bono, James L; Bosilevac, Joseph M

2014-10-01

Shiga toxin-producing Escherichia coli are a growing concern in the area of food safety, and the United States Department of Agriculture Food Safety and Inspection Service has identified the serotypes O26, O45, O103, O111, O121, O145, and O157 as adulterants in certain types of raw beef. The most relevant to human disease are the enterohemorrhagic E. coli (EHEC) strains that possess intimin (eae), Shiga toxin 1 and/or 2 (stx1-2), and in most cases the conserved pO157 or pO157 like virulence plasmid. Contamination of raw beef with EHEC is likely to occur via the transfer of cattle feces on hides to the carcass. To detect EHEC directly from cattle feces, we evaluated the utility of a multiplex real time PCR assay that targets the EHEC associated gene target ecf1 in combination with eae and stx1-2. Our assay had an increased sensitivity and provided a reliable limit of detection (LOD) of 1.25×10(3)colony-forming unitspermL (CFUs/mL) in an EHEC spiked fecal background. In addition, we evaluated the use of a duplex qPCR assay using ecf1 for the enumeration of total EHEC directly from cattle feces. The reliable limit of quantification (LOQ) was determined to be 1.25×10(3)CFUs/mL. Our assay requires minimal sample processing and provides LOD and LOQ of EHEC directly from cattle feces that are the lowest reported. The application of this assay towards the identification of cattle shedding EHEC at a level above 1.25×10(3)CFUs/mL could be a first line of defense in identifying cattle shedding these pathogens. Published by Elsevier B.V.

Acquisition and transmission of Theileria parva by vector tick, Rhipicephalus appendiculatus.

PubMed

Konnai, Satoru; Imamura, Saiki; Nakajima, Chie; Witola, William Harold; Yamada, Shinji; Simuunza, Martin; Nambota, Andrew; Yasuda, Jun; Ohashi, Kazuhiko; Onuma, Misao

2006-08-01

In order to investigate the transmission dynamics of Theileria parva (T. parva) by the brown ear tick, Rhipicephalus appendiculatus (R. appendiculatus), under experimental conditions, detection of T. parva in ticks and cattle was performed by a quantitative real-time PCR assay. A calf inoculated with a T. parva mixture became PCR-positive for T. parva infection on day 8 post-inoculation, and subsequently, nymphal ticks were introduced and maintained to feed on the infected calf for 6 days. Engorged nymphs were collected daily and allowed to molt into adults, and overall, 70.8% (121/171) of the adult ticks acquired the T. parva infection. Furthermore, the T. parva infection rate in ticks under field conditions was monitored by real-time PCR in R. appendiculatus ticks collected from a traditionally managed pastoral land of Zambia, on which Sanga breed cattle are traditionally reared and the area has endemic East Coast fever (ECF). A total of 70 cattle were randomly selected in the same area and 67 (95.7%) were found to be serologically positive for R. appendiculatus tick antigen (RIM36). Twenty-nine (43.3%) of the 67 serologically positive cattle were real-time PCR-positive for T. parva, although no piroplasms could be detected in the blood smears. Unexpectedly, out of 614 R. appendiculatus nymphal and adult ticks collected by flagging vegetation, 4.1% were positive for T. parva DNA. However, since the rate of transmission of T. parva from infected cattle to ticks and vice versa and the serological evidence of exposure to R. appendiculatus ticks in naturally exposed cattle were relatively high, it would be wise in such a case to consider vector control as well as vaccination against ECF as control measures.

Evaluation of the Microsoft Kinect as a clinical assessment tool of body sway.

PubMed

Yeung, L F; Cheng, Kenneth C; Fong, C H; Lee, Winson C C; Tong, Kai-Yu

2014-09-01

Total body center of mass (TBCM) is a useful kinematic measurement of body sway. However, expensive equipment and high technical requirement limit the use of motion capture systems in large-scale clinical settings. Center of pressure (CP) measurement obtained from force plates cannot accurately represent TBCM during large body sway movement. Microsoft Kinect is a rapidly developing, inexpensive, and portable posturographic device, which provides objective and quantitative measurement of TBCM sway. The purpose of this study was to evaluate Kinect as a clinical assessment tool for TBCM sway measurement. The performance of the Kinect system was compared with a Vicon motion capture system and a force plate. Ten healthy male subjects performed four upright quiet standing tasks: (1) eyes open (EOn), (2) eyes closed (ECn), (3) eyes open standing on foam (EOf), and (4) eyes closed standing on foam (ECf). Our results revealed that the Kinect system produced highly correlated measurement of TBCM sway (mean RMSE=4.38 mm; mean CORR=0.94 in Kinect-Vicon comparison), as well as comparable intra-session reliability to Vicon. However, the Kinect device consistently overestimated the 95% CL of sway by about 3mm. This offset could be due to the limited accuracy, resolution, and sensitivity of the Kinect sensors. The Kinect device was more accurate in the medial-lateral than in the anterior-posterior direction, and performed better than the force plate in more challenging balance tasks, such as (ECf) with larger TBCM sway. Overall, Kinect is a cost-effective alternative to a motion capture and force plate system for clinical assessment of TBCM sway. Copyright © 2014 Elsevier B.V. All rights reserved.

Revisiting atenolol as a low passive permeability marker.

PubMed

Chen, Xiaomei; Slättengren, Tim; de Lange, Elizabeth C M; Smith, David E; Hammarlund-Udenaes, Margareta

2017-10-31

Atenolol, a hydrophilic beta blocker, has been used as a model drug for studying passive permeability of biological membranes such as the blood-brain barrier (BBB) and the intestinal epithelium. However, the extent of S-atenolol (the active enantiomer) distribution in brain has never been evaluated, at equilibrium, to confirm that no transporters are involved in its transport at the BBB. To assess whether S-atenolol, in fact, depicts the characteristics of a low passive permeable drug at the BBB, a microdialysis study was performed in rats to monitor the unbound concentrations of S-atenolol in brain extracellular fluid (ECF) and plasma during and after intravenous infusion. A pharmacokinetic model was developed, based on the microdialysis data, to estimate the permeability clearance of S-atenolol into and out of brain. In addition, the nonspecific binding of S-atenolol in brain homogenate was evaluated using equilibrium dialysis. The steady-state ratio of unbound S-atenolol concentrations in brain ECF to that in plasma (i.e., K p,uu,brain ) was 3.5% ± 0.4%, a value much less than unity. The unbound volume of distribution in brain (V u, brain ) of S-atenolol was also calculated as 0.69 ± 0.10 mL/g brain, indicating that S-atenolol is evenly distributed within brain parenchyma. Lastly, equilibrium dialysis showed limited nonspecific binding of S-atenolol in brain homogenate with an unbound fraction (f u,brain ) of 0.88 ± 0.07. It is concluded, based on K p,uu,brain being much smaller than unity, that S-atenolol is actively effluxed at the BBB, indicating the need to re-consider S-atenolol as a model drug for passive permeability studies of BBB transport or intestinal absorption.

Role of O2 in the Growth of Rhizobium leguminosarum bv. viciae 3841 on Glucose and Succinate

PubMed Central

Wheatley, Rachel M.; Ramachandran, Vinoy K.; Geddes, Barney A.; Perry, Benjamin J.; Yost, Chris K.

2016-01-01

ABSTRACT Insertion sequencing (INSeq) analysis of Rhizobium leguminosarum bv. viciae 3841 (Rlv3841) grown on glucose or succinate at both 21% and 1% O2 was used to understand how O2 concentration alters metabolism. Two transcriptional regulators were required for growth on glucose (pRL120207 [eryD] and RL0547 [phoB]), five were required on succinate (pRL100388, RL1641, RL1642, RL3427, and RL4524 [ecfL]), and three were required on 1% O2 (pRL110072, RL0545 [phoU], and RL4042). A novel toxin-antitoxin system was identified that could be important for generation of new plasmidless rhizobial strains. Rlv3841 appears to use the methylglyoxal pathway alongside the Entner-Doudoroff (ED) pathway and tricarboxylic acid (TCA) cycle for optimal growth on glucose. Surprisingly, the ED pathway was required for growth on succinate, suggesting that sugars made by gluconeogenesis must undergo recycling. Altered amino acid metabolism was specifically needed for growth on glucose, including RL2082 (gatB) and pRL120419 (opaA, encoding omega-amino acid:pyruvate transaminase). Growth on succinate specifically required enzymes of nucleobase synthesis, including ribose-phosphate pyrophosphokinase (RL3468 [prs]) and a cytosine deaminase (pRL90208 [codA]). Succinate growth was particularly dependent on cell surface factors, including the PrsD-PrsE type I secretion system and UDP-galactose production. Only RL2393 (glnB, encoding nitrogen regulatory protein PII) was specifically essential for growth on succinate at 1% O2, conditions similar to those experienced by N2-fixing bacteroids. Glutamate synthesis is constitutively activated in glnB mutants, suggesting that consumption of 2-ketoglutarate may increase flux through the TCA cycle, leading to excess reductant that cannot be reoxidized at 1% O2 and cell death. IMPORTANCE Rhizobium leguminosarum, a soil bacterium that forms N2-fixing symbioses with several agriculturally important leguminous plants (including pea, vetch, and lentil), has been widely utilized as a model to study Rhizobium-legume symbioses. Insertion sequencing (INSeq) has been used to identify factors needed for its growth on different carbon sources and O2 levels. Identification of these factors is fundamental to a better understanding of the cell physiology and core metabolism of this bacterium, which adapts to a variety of different carbon sources and O2 tensions during growth in soil and N2 fixation in symbiosis with legumes. PMID:27795326

Transcriptome and membrane fatty acid analyses reveal different strategies for responding to permeating and non-permeating solutes in the bacterium Sphingomonas wittichii

PubMed Central

2011-01-01

Background Sphingomonas wittichii strain RW1 can completely oxidize dibenzo-p-dioxins and dibenzofurans, which are persistent contaminants of soils and sediments. For successful application in soil bioremediation systems, strain RW1 must cope with fluctuations in water availability, or water potential. Thus far, however, little is known about the adaptive strategies used by Sphingomonas bacteria to respond to changes in water potential. To improve our understanding, strain RW1 was perturbed with either the cell-permeating solute sodium chloride or the non-permeating solute polyethylene glycol with a molecular weight of 8000 (PEG8000). These solutes are assumed to simulate the solute and matric components of the total water potential, respectively. The responses to these perturbations were then assessed and compared using a combination of growth assays, transcriptome profiling, and membrane fatty acid analyses. Results Under conditions producing a similar decrease in water potential but without effect on growth rate, there was only a limited shared response to perturbation with sodium chloride or PEG8000. This shared response included the increased expression of genes involved with trehalose and exopolysaccharide biosynthesis and the reduced expression of genes involved with flagella biosynthesis. Mostly, the responses to perturbation with sodium chloride or PEG8000 were very different. Only sodium chloride triggered the increased expression of two ECF-type RNA polymerase sigma factors and the differential expression of many genes involved with outer membrane and amino acid metabolism. In contrast, only PEG8000 triggered the increased expression of a heat shock-type RNA polymerase sigma factor along with many genes involved with protein turnover and repair. Membrane fatty acid analyses further corroborated these differences. The degree of saturation of membrane fatty acids increased after perturbation with sodium chloride but had the opposite effect and decreased after perturbation with PEG8000. Conclusions A combination of growth assays, transcriptome profiling, and membrane fatty acid analyses revealed that permeating and non-permeating solutes trigger different adaptive responses in strain RW1, suggesting these solutes affect cells in fundamentally different ways. Future work is now needed that connects these responses with the responses observed in more realistic scenarios of soil desiccation. PMID:22082453

Analytical transport network theory to guide the design of 3-D microstructural networks in energy materials: Part 1. Flow without reactions

NASA Astrophysics Data System (ADS)

Cocco, Alex P.; Nakajo, Arata; Chiu, Wilson K. S.

2017-12-01

We present a fully analytical, heuristic model - the "Analytical Transport Network Model" - for steady-state, diffusive, potential flow through a 3-D network. Employing a combination of graph theory, linear algebra, and geometry, the model explicitly relates a microstructural network's topology and the morphology of its channels to an effective material transport coefficient (a general term meant to encompass, e.g., conductivity or diffusion coefficient). The model's transport coefficient predictions agree well with those from electrochemical fin (ECF) theory and finite element analysis (FEA), but are computed 0.5-1.5 and 5-6 orders of magnitude faster, respectively. In addition, the theory explicitly relates a number of morphological and topological parameters directly to the transport coefficient, whereby the distributions that characterize the structure are readily available for further analysis. Furthermore, ATN's explicit development provides insight into the nature of the tortuosity factor and offers the potential to apply theory from network science and to consider the optimization of a network's effective resistance in a mathematically rigorous manner. The ATN model's speed and relative ease-of-use offer the potential to aid in accelerating the design (with respect to transport), and thus reducing the cost, of energy materials.

An outbreak of East Coast fever in a herd of Sanga cattle in Lutale, Central Province of Zambia.

PubMed

Minjauw, B; Otte, M J; James, A D; de Castro, J J; Permin, A; Di Giulo, G

1998-05-01

An outbreak of East Coast fever (ECF) occurred in an experimental herd of Sanga cattle maintained under a traditional rangeland grazing system at Lutale, Central Province of Zambia. Two groups of cattle had been kept under different tick-control regimens for several years prior to the introduction of the disease and epidemiological information on the outbreak were recorded. Weekly tick control was no sufficient to achieve full protection against Theileria parva infection. Systematic body temperature monitoring seems to be a good method for early detection of infection resulting in an important reduction of the case fatality rate after treatment with anti-theilerial drugs.

Hanford Soil Inventory Model (SIM-v2) Calculated Radionuclide Inventory of Direct Liquid Discharges to Soil in the Hanford Site's 200 Areas.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nichols, William E.; Zaher, U.; Agnew, S.

The Hanford soil inventory model (SIM) provides the basic radionuclide and chemical soil inventories from historical liquid discharges to about 400 sites at the Hanford Site. Although liquid discharge inventory for chemicals is part of the SIM implementation, only radionuclide inventory is discussed here since the focus of this ECF is on providing radionuclides inputs for the composite analysis (CA) per DOE Order 435.1, Radioactive Waste Management, requirements. Furthermore, discharged inventories are only estimated for the soluble portions of the liquid discharges to waste sites/waste management areas located on the 200 Area of the Hanford Site (Central Plateau).

Dynamic relationship between neurostimulation and N-acetylaspartate metabolism in the human visual cortex: evidence that NAA functions as a molecular water pump during visual stimulation.

PubMed

Baslow, Morris H; Hrabe, Jan; Guilfoyle, David N

2007-01-01

N-acetyl-l-aspartic acid (NAA), an amino acid synthesized and stored primarily in neurons in the brain, has been proposed to be a molecular water pump (MWP) whose function is to rapidly remove water from neurons against a water gradient. In this communication, we describe the results of a functional (1)H proton magnetic resonance spectroscopy (fMRS) study, and provide evidence that in the human visual cortex, over a 10-min period of visual stimulation, there are stimulation-induced graded changes in the NAA MRS signal from that of a preceding 10-min baseline period with a decline in the NAA signal of 13.1% by the end of the 10-min stimulation period. Upon cessation of visual stimulation, the NAA signal gradually increases during a 10-min recovery period and once again approaches the baseline level. Because the NAA MRS signal reflects the NAA concentration, these changes indicate rapid focal changes in its concentration, and transient changes in its intercompartmental metabolism. These include its rates of synthesis and efflux from neurons and its hydrolysis by oligodendrocytes. During stimulation, the apparent rate of NAA efflux and hydrolysis increased 14.2 times, from 0.55 to 7.8 micromol g(-1) h(-1). During recovery, the apparent rate of synthesis increased 13.3 times, from 0.55 to 7.3 micromol g(-1) h(-1). The decline in the NAA signal during stimulation suggests that a rapid increase in the rate of NAA-obligated water release to extracellular fluid (ECF) is the initial and seminal event in response to neurostimulation. It is concluded that the NAA metabolic cycle in the visual cortex is intimately linked to rates of neuronal signaling, and that the functional cycle of NAA is associated with its release to ECF, thus supporting the hypothesis that an important function of the NAA metabolic cycle is that of an efflux MWP.

Umbilical cord clamping in term piglets: a useful model to study perinatal asphyxia?

PubMed

van Dijk, A J; van Loon, J P A M; Taverne, M A M; Jonker, F H

2008-09-01

Perinatal asphyxia results in tissue and cellular changes during the reperfusion period and clinical signs like perinatal mortality and decreased vitality at birth in newborn piglets. This study aimed to develop and validate a model of birth asphyxia, mimicking the evolvement of birth asphyxia in natural farrowings by conducting umbilical cord clamping (UCC) in term piglets during caesarean sections under general anaesthesia. In total 23 piglets were subjected to 5-8min of UCC and 24 piglets served as controls. Acid-base balance values and heart rates measured before UCC remained fairly constant throughout the surgical procedure, indicating nearly identical starting conditions of piglets within and between litters. UCC resulted in a significant, mild, mixed respiratory-metabolic acidosis (pH 7.22, pCO(2) 9.8kPa, BE(ecf) 2mmol/L, lactate 6.5mmol/L; controls: pH 7.31, pCO(2) 8.5kPa, BE(ecf) 5mmol/L, lactate 4mmol/L) at 10min after birth (defined as simultaneous cutting of the umbilical cord and removal of a plastic bag that had been placed over the head to avoid air intake). Heart rates were significantly decreased during UCC (range: 83-107beats/min versus 128-134beats/min in controls). Rectal temperatures and changes in body weight until 72h of life were not affected by UCC. Interestingly, four control and seven clamped piglets did not survive as no independent respiration could be attained. Birth weights and duration of UCC of these piglets did not differ significantly from those in surviving control and clamped piglets. In conclusion the mixed respiratory-metabolic acidosis arising in the surviving clamped piglets is not as severe as can be expected in highly asphyxiated, vaginally delivered newborn piglets. Repeatability of the model is compromised by considerable variation in the individual response to UCC.

Genes encoding two Theileria parva antigens recognized by CD8+ T-cells exhibit sequence diversity in South Sudanese cattle populations but the majority of alleles are similar to the Muguga component of the live vaccine cocktail

PubMed Central

Pelle, Roger; Mwacharo, Joram M.; Njahira, Moses N.; Marcellino, Wani L.; Kiara, Henry; Malak, Agol K.; EL Hussein, Abdel Rahim M.; Bishop, Richard; Skilton, Robert A.

2017-01-01

East Coast fever (ECF), caused by Theileria parva infection, is a frequently fatal disease of cattle in eastern, central and southern Africa, and an emerging disease in South Sudan. Immunization using the infection and treatment method (ITM) is increasingly being used for control in countries affected by ECF, but not yet in South Sudan. It has been reported that CD8+ T-cell lymphocytes specific for parasitized cells play a central role in the immunity induced by ITM and a number of T. parva antigens recognized by parasite-specific CD8+ T-cells have been identified. In this study we determined the sequence diversity among two of these antigens, Tp1 and Tp2, which are under evaluation as candidates for inclusion in a sub-unit vaccine. T. parva samples (n = 81) obtained from cattle in four geographical regions of South Sudan were studied for sequence polymorphism in partial sequences of the Tp1 and Tp2 genes. Eight positions (1.97%) in Tp1 and 78 positions (15.48%) in Tp2 were shown to be polymorphic, giving rise to four and 14 antigen variants in Tp1 and Tp2, respectively. The overall nucleotide diversity in the Tp1 and Tp2 genes was π = 1.65% and π = 4.76%, respectively. The parasites were sampled from regions approximately 300 km apart, but there was limited evidence for genetic differentiation between populations. Analyses of the sequences revealed limited numbers of amino acid polymorphisms both overall and in residues within the mapped CD8+ T-cell epitopes. Although novel epitopes were identified in the samples from South Sudan, a large number of the samples harboured several epitopes in both antigens that were similar to those in the T. parva Muguga reference stock, which is a key component in the widely used live vaccine cocktail. PMID:28231338

Bleach boosting effect of xylanase A from Bacillus halodurans C-125 in ECF bleaching of wheat straw pulp.

PubMed

Lin, Xiao-qiong; Han, Shuang-yan; Zhang, Na; Hu, Hui; Zheng, Sui-ping; Ye, Yan-rui; Lin, Ying

2013-02-05

Past studies have revealed major difficulties in applications of xylanase in the pulp and paper industry as enzymes isolated from many different species could not tolerate high temperatures or highly alkaline conditions. The thermostable xylanase A from Bacillus halodurans C-125 (C-125 xylanase A) was successfully cloned and expressed in Pichia pastoris with a yield as high as 3361 U/mL in a 2 L reactor. Its thermophilic and basophilic properties (optimal activity at 70 °C and pH 9.0), together with the fact it is cellulase-free, render this enzyme attractive for compatible applications in the pulp and paper industry. The pretreatment of wheat straw pulp with C-125 xylanase A at pH 9.0 and 70 °C for 90 min induced the release of both chromophores (Ab(237), Ab(254), Ab(280)) and hydrophobic compounds (Ab(465)) into the filtrate as well as sugar degradation. Moreover, the addition of 10 U xylanase to 1 g wheat straw pulp (dry weight) as pretreatment improved brightness by 5.2% ISO and decreased the kappa number by 5.0% when followed by hydrogen peroxide bleaching. In addition, compared with two commercial enzymes, Pulpzyme HC and AU-PE89, which are normally incorporated in ECF bleaching of wheat straw pulp, C-125 xylanase A proved to be more effective in enhancing brightness as well as preserving paper strength properties. When evaluating the physical properties of pulp samples, such as tensile index, tearing index, bursting index, and post-color (PC) number, the enzymes involved in pretreating pulps exhibited better or the same performances as chemical treatment. Compared with chemical bleaching, chlorine consumption can be significantly reduced by 10% for xylanase-pretreated wheat straw pulp while maintaining the brightness together with the kappa number at the same level. Scanning electron microscopy revealed significant surface modification of enzyme-pretreated pulp fibers with no marked fiber disruptions. Copyright © 2012 Elsevier Inc. All rights reserved.

Anticancer effects of brominated indole alkaloid Eudistomin H from marine ascidian Eudistoma viride against cervical cancer cells (HeLa).

PubMed

Rajesh, Rajaian Pushpabai; Annappan, Murugan

2015-01-01

Marine invertebrates called ascidians are prolific producers of bioactive substances. The ascidian Eudistoma viride, distributed along the Southeast coast of India, was investigated for its in vitro cytotoxic activity against human cervical carcinoma (HeLa) cells by the MTT assay. The crude methanolic extract of E. viride, with an IC50 of 53 μg/ml, was dose-dependently cytotoxic. It was more potent at 100 μg/ml than cyclohexamide (1 μg/ml), reducing cell viability to 9.2%. Among nine fractions separated by chromatography, ECF-8 exhibited prominent cytoxic activity at 10 μg/ml. The HPLC fraction EHF-21 of ECF-8 was remarkably dose- and time-dependently cytotoxic, with 39.8% viable cells at 1 μg/ml compared to 51% in cyclohexamide-treated cells at the same concentration; the IC50 was 0.49 μg/ml. Hoechst staining of HeLa cells treated with EHF-21 at 0.5 μg/ml revealed apoptotic events such an cell shrinkage, membrane blebbing, chromatin condensation and formation of apoptotic bodies. Cell size and granularity study showed changes in light scatter, indicating the characteristic feature of cells dying by apoptosis. The cell-cycle analysis of HeLa cells treated with fraction EHF-21 at 1 μg/ml showed the marked arrest of cells in G0/G1, S and G2/M phases and an increase in the sub G0/G1 population indicated an increase in the apoptotic cell population. The statistical analysis of the sub-G1 region showed a dose-dependent induction of apoptosis. DNA fragmentation was also observed in HeLa cells treated with EHF-21. The active EHF-21 fraction, a brominated indole alkaloid Eudistomin H, led to apoptotic death of HeLa cells. Copyright© 2015 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.

Effects of evaporative cooling on the regulation of body water and milk production in crossbred Holstein cattle in a tropical environment

NASA Astrophysics Data System (ADS)

Chaiyabutr, N.; Chanpongsang, S.; Suadsong, S.

2008-09-01

The aim of this study was to determine how evaporative cooling modifies body function with respect to water metabolism and other variables relevant to milk synthesis in crossbred cattle. The study was conducted on two groups of 0.875HF:0.125RS crossbred Holstein cattle (87.5%) housed in an open-sided barn with a tiled roof (non-cooled animals) and in a close-sided barn under an evaporative cooling system (cooled animals). The maximum ambient temperature and relative humidity for the non-cooled group were 33°C and 61%, with the corresponding values for the evaporatively cooled barn being 28°C and 84%, respectively. The temperature humidity index (THI) of under non-cooled conditions was higher ( P < 0.05) than that in the cooled barn. Rectal temperatures and respiration rates of non-cooled animals were higher ( P < 0.05) than those of cooled animals. Daily dry matter intake (DMI) of cooled animals was higher while water intakes were lower ( P < 0.05) than those of non-cooled animals. The mean absolute values of plasma volume, blood volume, and extracellular fluid (ECF) of cooled animals were significantly higher ( P < 0.05) than those of non-cooled animals throughout all stages of lactation. Milk yields of cooled animals were higher by 42%, 36% and 79% on average than those of non-cooled animals during early-, mid- and late-lactation, respectively. The decline in milk yields as lactation advances was markedly apparent in late-lactating non-cooled animals, while no significant changes in milk composition at different stages of lactation were observed in either group. Mean arterial plasma concentrations, arteriovenous concentration differences (A-V differences) and the extraction ratio across the mammary gland for acetate, glucose and triglyceride of cooled animals were not significantly different compared with values for non-cooled animals. No differences were seen in plasma hormonal levels for triiodotyronine (T3) and insulin-like growth factor-1 (IGF-1), but plasma cortisol and thyroxine (T4) levels tended to be lower in non-cooled animals. This study suggests that low cooling temperature accompanied by high humidity influences a galactopoietic effect, in part through increases in ECF, blood volume and plasma volume in association with an increase in DMI, which partitions the distribution of nutrients to the mammary gland for milk synthesis. Cooled animals were unable to maintain high milk yield as lactation advances even though a high level of body fluids was maintained during long-term cooled exposure. The decline in milk yield, coinciding with a decrease in net energy for lactation as lactation advances, could be attributed to a local change within the mammary gland.

Whole-genome phylogenies of the family Bacillaceae and expansion of the sigma factor gene family in the Bacillus cereus species-group

PubMed Central

2011-01-01

Background The Bacillus cereus sensu lato group consists of six species (B. anthracis, B. cereus, B. mycoides, B. pseudomycoides, B. thuringiensis, and B. weihenstephanensis). While classical microbial taxonomy proposed these organisms as distinct species, newer molecular phylogenies and comparative genome sequencing suggests that these organisms should be classified as a single species (thus, we will refer to these organisms collectively as the Bc species-group). How do we account for the underlying similarity of these phenotypically diverse microbes? It has been established for some time that the most rapidly evolving and evolutionarily flexible portions of the bacterial genome are regulatory sequences and transcriptional networks. Other studies have suggested that the sigma factor gene family of these organisms has diverged and expanded significantly relative to their ancestors; sigma factors are those portions of the bacterial transcriptional apparatus that control RNA polymerase recognition for promoter selection. Thus, examining sigma factor divergence in these organisms would concurrently examine both regulatory sequences and transcriptional networks important for divergence. We began this examination by comparison to the sigma factor gene set of B. subtilis. Results Phylogenetic analysis of the Bc species-group utilizing 157 single-copy genes of the family Bacillaceae suggests that several taxonomic revisions of the genus Bacillus should be considered. Within the Bc species-group there is little indication that the currently recognized species form related sub-groupings, suggesting that they are members of the same species. The sigma factor gene family encoded by the Bc species-group appears to be the result of a dynamic gene-duplication and gene-loss process that in previous analyses underestimated the true heterogeneity of the sigma factor content in the Bc species-group. Conclusions Expansion of the sigma factor gene family appears to have preferentially occurred within the extracytoplasmic function (ECF) sigma factor genes, while the primary alternative (PA) sigma factor genes are, in general, highly conserved with those found in B. subtilis. Divergence of the sigma-controlled transcriptional regulons among various members of the Bc species-group likely has a major role in explaining the diversity of phenotypic characteristics seen in members of the Bc species-group. PMID:21864360

Virtual reality robotic telesurgery simulations using MEMICA haptic system

NASA Technical Reports Server (NTRS)

Bar-Cohen, Yoseph; Mavroidis, Constantinos; Bouzit, Mourad; Dolgin, Benjamin; Harm, Deborah L.; Kopchok, George E.; White, Rodney

2001-01-01

The authors conceived a haptic mechanism called MEMICA (Remote Mechanical Mirroring using Controlled stiffness and Actuators) that can enable the design of high dexterity, rapid response, and large workspace haptic system. The development of a novel MEMICA gloves and virtual reality models are being explored to allow simulation of telesurgery and other applications. The MEMICA gloves are being designed to provide intuitive mirroring of the conditions at a virtual site where a robot simulates the presence of a human operator. The key components of MEMICA are miniature electrically controlled stiffness (ECS) elements and electrically controlled force and stiffness (ECFS) actuators that are based on the use of Electro-Rheological Fluids (ERF. In this paper the design of the MEMICA system and initial experimental results are presented.

The effects of chemical coagulants on the decolorization of dyes by electrocoagulation using response surface methodology (RSM)

NASA Astrophysics Data System (ADS)

Butler, Erick B.; Hung, Yung-Tse; Mulamba, Oliver

2017-09-01

This study assessed the efficiency of electrocoagulation (ECF) coupled with an addition of chemical coagulant to decolorize textile dye. Tests were conducted using Box Behnken methodology to vary six parameters: dye type, weight, coagulant type, dose, initial pH and current density. The combination of electrocoagulation and chemical coagulation was able to decolorize dye up to 99.42 % in 30 min of treatment time which is remarkably shorter in comparison with using conventional chemical coagulation. High color removal was found to be contingent upon the dye type and current density, along with the interactions between the current density and the coagulant dose. The addition of chemical coagulants did enhanced treatment efficiency.

Power Systems Life Cycle Analysis Tool (Power L-CAT).

DOE Office of Scientific and Technical Information (OSTI.GOV)

Andruski, Joel; Drennen, Thomas E.

2011-01-01

The Power Systems L-CAT is a high-level dynamic model that calculates levelized production costs and tracks environmental performance for a range of electricity generation technologies: natural gas combined cycle (using either imported (LNGCC) or domestic natural gas (NGCC)), integrated gasification combined cycle (IGCC), supercritical pulverized coal (SCPC), existing pulverized coal (EXPC), nuclear, and wind. All of the fossil fuel technologies also include an option for including carbon capture and sequestration technologies (CCS). The model allows for quick sensitivity analysis on key technical and financial assumptions, such as: capital, O&M, and fuel costs; interest rates; construction time; heat rates; taxes; depreciation;more » and capacity factors. The fossil fuel options are based on detailed life cycle analysis reports conducted by the National Energy Technology Laboratory (NETL). For each of these technologies, NETL's detailed LCAs include consideration of five stages associated with energy production: raw material acquisition (RMA), raw material transport (RMT), energy conversion facility (ECF), product transportation and distribution (PT&D), and end user electricity consumption. The goal of the NETL studies is to compare existing and future fossil fuel technology options using a cradle-to-grave analysis. The NETL reports consider constant dollar levelized cost of delivered electricity, total plant costs, greenhouse gas emissions, criteria air pollutants, mercury (Hg) and ammonia (NH3) emissions, water withdrawal and consumption, and land use (acreage).« less

The global security perspective on the effects of executive cognitive function on complex behavioral screening intervention and HIV/AIDS.

PubMed

Kim, Suk-Hee

2010-11-01

The purpose of this quantitative study is to understand the global security perspective on the effects of executive cognitive function (ECF) on Complex Behavioral Screening Intervention and HIV/AIDS. The HIV/AIDS pandemic is as much a social, political, economic, and cultural problem as a biomedical one. HIV/AIDS is associated centrally with the collapse not just of communities and families but potentially of states, with some of the largest public health interventions ever and enormous questions about governance, a huge population of orphans, and deep questions about intergenerational relations and cultural transmission. This study also is to develop a screening instrument that improves quality of life for individuals with executive cognitive impairments and behavior problems in our communities and the global society.

An extracytoplasmic function sigma factor-dependent periplasmic glutathione peroxidase is involved in oxidative stress response of Shewanella oneidensis

DOE PAGES

Dai, Jingcheng; Wei, Hehong; Tian, Chunyuan; ...

2015-01-01

Background: Bacteria use alternative sigma factors (σs) to regulate condition-specific gene expression for survival and Shewanella harbors multiple ECF (extracytoplasmic function) σ genes and cognate anti-sigma factor genes. Here we comparatively analyzed two of the rpoE-like operons in the strain MR-1: rpoE-rseA-rseB-rseC and rpoE2-chrR. Results: RpoE was important for bacterial growth at low and high temperatures, in the minimal medium, and high salinity. The degP/htrA orthologue, required for growth of Escherichia coli and Pseudomonas aeruginosa at high temperature, is absent in Shewanella, while the degQ gene is RpoE-regulated and is required for bacterial growth at high temperature. RpoE2 was essentialmore » for the optimal growth in oxidative stress conditions because the rpoE2 mutant was sensitive to hydrogen peroxide and paraquat. The operon encoding a ferrochelatase paralogue (HemH2) and a periplasmic glutathione peroxidase (PgpD) was identified as RpoE2-dependent. PgpD exhibited higher activities and played a more important role in the oxidative stress responses than the cytoplasmic glutathione peroxidase CgpD under tested conditions. The rpoE2-chrR operon and the identified regulon genes, including pgpD and hemH2, are coincidently absent in several psychrophilic and/or deep-sea Shewanella strains. Conclusion: In S. oneidensis MR-1, the RpoE-dependent degQ gene is required for optimal growth under high temperature. The rpoE2 and RpoE2-dependent pgpD gene encoding a periplasmic glutathione peroxidase are involved in oxidative stress responses. But rpoE2 is not required for bacterial growth at low temperature and it even affected bacterial growth under salt stress, indicating that there is a tradeoff between the salt resistance and RpoE2-mediated oxidative stress responses.« less

An extracytoplasmic function sigma factor-dependent periplasmic glutathione peroxidase is involved in oxidative stress response of Shewanella oneidensis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dai, Jingcheng; Wei, Hehong; Tian, Chunyuan

Background: Bacteria use alternative sigma factors (σs) to regulate condition-specific gene expression for survival and Shewanella harbors multiple ECF (extracytoplasmic function) σ genes and cognate anti-sigma factor genes. Here we comparatively analyzed two of the rpoE-like operons in the strain MR-1: rpoE-rseA-rseB-rseC and rpoE2-chrR. Results: RpoE was important for bacterial growth at low and high temperatures, in the minimal medium, and high salinity. The degP/htrA orthologue, required for growth of Escherichia coli and Pseudomonas aeruginosa at high temperature, is absent in Shewanella, while the degQ gene is RpoE-regulated and is required for bacterial growth at high temperature. RpoE2 was essentialmore » for the optimal growth in oxidative stress conditions because the rpoE2 mutant was sensitive to hydrogen peroxide and paraquat. The operon encoding a ferrochelatase paralogue (HemH2) and a periplasmic glutathione peroxidase (PgpD) was identified as RpoE2-dependent. PgpD exhibited higher activities and played a more important role in the oxidative stress responses than the cytoplasmic glutathione peroxidase CgpD under tested conditions. The rpoE2-chrR operon and the identified regulon genes, including pgpD and hemH2, are coincidently absent in several psychrophilic and/or deep-sea Shewanella strains. Conclusion: In S. oneidensis MR-1, the RpoE-dependent degQ gene is required for optimal growth under high temperature. The rpoE2 and RpoE2-dependent pgpD gene encoding a periplasmic glutathione peroxidase are involved in oxidative stress responses. But rpoE2 is not required for bacterial growth at low temperature and it even affected bacterial growth under salt stress, indicating that there is a tradeoff between the salt resistance and RpoE2-mediated oxidative stress responses.« less

Immunization against East Coast Fever in field cattle with low infectivity Theileria parva stabilate--preliminary assessment.

PubMed

Mbassa, G K; Kweka, L E; Dulla, P N

1998-05-01

Two Theileria parva sporozoite stabilates stored at -196 degrees C, then at -70 degrees C for six weeks (stabilate 1) and more than six months (stabilate 2) were inoculated into four eight-month old male calves, 1 and 2 (stabilate 1), and 3 and 4 (stabilate 2). Calves 1 and 2 developed pyrexia, enlargement of lymph nodes, and the former died of East Coast Fever. Calves 3 and 4 showed slight enlargement of lymph nodes without fever. Lymph node smears from all calves (from day 10 to 20 post-inoculation) showed lymphoblasts, phagocytic macrophages, and schizonts. Piroplasms were detected in erythrocytes in blood smears from calves 1 and 2 but not in calves 3 and 4. Calves 2, 3 and 4 recovered without any treatment while calf 1 died of East Coast Fever on day 20. Serum samples from recovered calves taken on day 30 of the experiment were positive for antischizont antibodies to T. parva at 1:640 dilution, but pre-inoculation serum samples were negative. Stabilate 2 was used to immunize 64 Boran, Friesian, Ayrshire and crosses with Zebu cattle in four herds with 25% reduction of oxytetracycline dose. All the animals except one calf recovered without any severe reactions. The latter died of disease other than ECF after the monitoring period was over (day 24). Day 30 post-inoculation serum samples were positive for T. parva antischizont antibodies. A follow-up of the remaining animals for over one year revealed no further ECF incidences in these herds. This experiment shows the loss of infectivity of the vaccine stored at temperatures higher than -196 degrees C. dependent on the duration. However, despite the lack of clinical signs in calves 3 and 4, there was cellular response and antibody production, and the stabilate for vaccine against East Coast fever can thus be stored prior to use at higher than -196 degrees C and still maintain capability to produce antibodies in field cattle, eliminating the use of oxytetracycline and monitoring. The vaccine will be cheaper and easier to use and the requirement for liquid nitrogen in the field reduced and the scale of application of the vaccine widened.

TpUB05, a Homologue of the Immunodominant Plasmodium falciparum Protein UB05, Is a Marker of Protective Immune Responses in Cattle Experimentally Vaccinated against East Coast Fever

PubMed Central

Dinga, Jerome Nyhalah; Wamalwa, Mark; Njimoh, Dieudonné Lemuh; Njahira, Moses N.; Djikeng, Appolinaire; Skilton, Rob; Titanji, Vincent Pryde Kehdingha; Pellé, Roger

2015-01-01

Introduction East Coast fever, a devastating disease of cattle, can be controlled partially by vaccination with live T. parva sporozoites. The antigens responsible for conferring immunity are not fully characterized. Recently it was shown that the P. falciparum immunodominant protein UB05 is highly conserved in T. parva, the causative agent of East Coast fever. The aim of the present investigation was to determine the role of the homologue TpUB05 in protective immunity to East Coast fever. Methods The cloning, sequencing and expression of TpUB05 were done according to standard protocols. Bioinformatics analysis of TpUB05 gene was carried out using algorithms found in the public domain. Polyclonal antiserum against recombinant TpUB05 were raised in rabbits and used for further analysis by Western blotting, ELISA, immunolocalization and in vitro infection neutralization assay. The ability of recombinant TpUB05 (r-TpUB05) to stimulate bovine PBMCs ex-vivo to produce IFN-γ or to proliferate was tested using ELISpot and [3H]-thymidine incorporation assays, respectively. Results All the 20 cattle immunised by the infection and treatment method (ITM) developed significantly higher levels of TpUB05 specific antibodies (p<0.0001) compared to the non-vaccinated ones. Similarly, r-TpUB05 highly stimulated bovine PMBCs from 8/12 (67%) of ITM-immunized cattle tested to produce IFN-γ and proliferate (p< 0.029) as compared to the 04 naїve cattle included as controls. Polyclonal TpUB05 antiserum raised against r-TpUB05 also marginally inhibited infection (p < 0.046) of bovine PBMCs by T. parva sporozoites. In further experiments RT-PCR showed that the TpUB05 gene is expressed by the parasite. This was confirmed by immunolocalization studies which revealed TpUB05 expression by schizonts and piroplasms. Bioinformatics analysis also revealed that this antigen possesses two transmembrane domains, a N-glycosylation site and several O-glycosylation sites. Conclusion It was concluded that TpUB05 is a potential marker of protective immunity in ECF worth investigating further. PMID:26053064

TpUB05, a Homologue of the Immunodominant Plasmodium falciparum Protein UB05, Is a Marker of Protective Immune Responses in Cattle Experimentally Vaccinated against East Coast Fever.

PubMed

Dinga, Jerome Nyhalah; Wamalwa, Mark; Njimoh, Dieudonné Lemuh; Njahira, Moses N; Djikeng, Appolinaire; Skilton, Rob; Titanji, Vincent Pryde Kehdingha; Pellé, Roger

2015-01-01

East Coast fever, a devastating disease of cattle, can be controlled partially by vaccination with live T. parva sporozoites. The antigens responsible for conferring immunity are not fully characterized. Recently it was shown that the P. falciparum immunodominant protein UB05 is highly conserved in T. parva, the causative agent of East Coast fever. The aim of the present investigation was to determine the role of the homologue TpUB05 in protective immunity to East Coast fever. The cloning, sequencing and expression of TpUB05 were done according to standard protocols. Bioinformatics analysis of TpUB05 gene was carried out using algorithms found in the public domain. Polyclonal antiserum against recombinant TpUB05 were raised in rabbits and used for further analysis by Western blotting, ELISA, immunolocalization and in vitro infection neutralization assay. The ability of recombinant TpUB05 (r-TpUB05) to stimulate bovine PBMCs ex-vivo to produce IFN-γ or to proliferate was tested using ELISpot and [3H]-thymidine incorporation assays, respectively. All the 20 cattle immunised by the infection and treatment method (ITM) developed significantly higher levels of TpUB05 specific antibodies (p<0.0001) compared to the non-vaccinated ones. Similarly, r-TpUB05 highly stimulated bovine PMBCs from 8/12 (67%) of ITM-immunized cattle tested to produce IFN-γ and proliferate (p< 0.029) as compared to the 04 naїve cattle included as controls. Polyclonal TpUB05 antiserum raised against r-TpUB05 also marginally inhibited infection (p < 0.046) of bovine PBMCs by T. parva sporozoites. In further experiments RT-PCR showed that the TpUB05 gene is expressed by the parasite. This was confirmed by immunolocalization studies which revealed TpUB05 expression by schizonts and piroplasms. Bioinformatics analysis also revealed that this antigen possesses two transmembrane domains, a N-glycosylation site and several O-glycosylation sites. It was concluded that TpUB05 is a potential marker of protective immunity in ECF worth investigating further.

OGUPSA sensor scheduling architecture and algorithm

NASA Astrophysics Data System (ADS)

Zhang, Zhixiong; Hintz, Kenneth J.

1996-06-01

This paper introduces a new architecture for a sensor measurement scheduler as well as a dynamic sensor scheduling algorithm called the on-line, greedy, urgency-driven, preemptive scheduling algorithm (OGUPSA). OGUPSA incorporates a preemptive mechanism which uses three policies, (1) most-urgent-first (MUF), (2) earliest- completed-first (ECF), and (3) least-versatile-first (LVF). The three policies are used successively to dynamically allocate and schedule and distribute a set of arriving tasks among a set of sensors. OGUPSA also can detect the failure of a task to meet a deadline as well as generate an optimal schedule in the sense of minimum makespan for a group of tasks with the same priorities. A side benefit is OGUPSA's ability to improve dynamic load balance among all sensors while being a polynomial time algorithm. Results of a simulation are presented for a simple sensor system.

Classical conditioning leads to changes in extracellular concentrations of ependymin in goldfish brain.

PubMed

Shashoua, V E; Hesse, G W

1989-04-10

ELISA measurements showed that brain extracellular fluid (ECF) levels of ependymin decreased for animals that learned to associate a paired presentation of a light stimulus (CS) with the onset of an electric shock (US), whereas no changes were obtained for control goldfish that received the same number of stimuli delivered in a random unpaired order. Studies of the time course of the changes showed an immediate decrease (19%) after training followed by an increase (20%) above baseline by 5 h and a final return to baseline by 25 h. These data extend the findings of previous experiments, which demonstrated a role for ependymin in two training procedures that involved motor learning, to classical conditioning where no motor learning occurs. Thus it appears that ependymin may have a functional role in molecular mechanisms of learning and memory in general.

Sources of non-fossil-fuel emissions in carbonaceous aerosols during early winter in Chinese cities

NASA Astrophysics Data System (ADS)

Liu, Di; Li, Jun; Cheng, Zhineng; Zhong, Guangcai; Zhu, Sanyuan; Ding, Ping; Shen, Chengde; Tian, Chongguo; Chen, Yingjun; Zhi, Guorui; Zhang, Gan

2017-09-01

China experiences frequent and severe haze outbreaks from the beginning of winter. Carbonaceous aerosols are regarded as an essential factor in controlling the formation and evolution of haze episodes. To elucidate the carbon sources of air pollution, source apportionment was conducted using radiocarbon (14C) and unique molecular organic tracers. Daily 24 h PM2. 5 samples were collected continuously from October 2013 to November 2013 in 10 Chinese cities. The 14C results indicated that non-fossil-fuel (NF) emissions were predominant in total carbon (TC; average = 65 ± 7 %). Approximately half of the EC was derived primarily from biomass burning (BB) (average = 46 ± 11 %), while over half of the organic carbon (OC) fraction comprised NF (average = 68 ± 7 %). On average, the largest contributor to TC was NF-derived secondary OC (SOCnf), which accounted for 46 ± 7 % of TC, followed by SOC derived from fossil fuels (FF) (SOCf; 16 ± 3 %), BB-derived primary OC (POCbb; 13 ± 5 %), POC derived from FF (POCf; 12 ± 3 %), EC derived from FF (ECf; 7 ± 2 %) and EC derived from BB (ECbb; 6 ± 2 %). The regional background carbonaceous aerosol composition was characterized by NF sources; POCs played a major role in northern China, while SOCs contributed more in other regions. However, during haze episodes, there were no dramatic changes in the carbon source or composition in the cities under study, but the contribution of POC from both FF and NF increased significantly.

Tumor response evaluation after neoadjuvant chemotherapy in locally advanced gastric adenocarcinoma: a prospective, multi-center cohort study

PubMed Central

De Martini, Paolo; Ceresoli, Marco; Mari, Giulio M.; Costanzi, Andrea; Maggioni, Dario; Pugliese, Raffaele; Ferrari, Giovanni

2017-01-01

Background To verify the prognostic value of the pathologic and radiological tumor response after neoadjuvant chemotherapy in the treatment of locally advanced gastric adenocarcinoma. Methods A total of 67 patients with locally advanced gastric cancer (clinical ≥ T2 or nodal disease and without evidence of distant metastases) underwent perioperative chemotherapy (ECF or ECX regimen) from December 2009 through June 2015 in two surgical units. Histopathological and radiological response to chemotherapy were evaluated by using tumor regression grade (TRG) (Becker’s criteria) and volume change assessed by CT. Results Fifty-one (86%) patients completed all chemotherapy scheduled cycles successfully and surgery was curative (R0) in 64 (97%) subjects. The histopathological analysis showed 19 (29%) specimens with TRG1 (less than 10% of vital tumor left) and 25 (37%) patients had partial or complete response (CR) assessed by CT scan. Median disease free survival (DFS) and overall survival (OS) were 25.70 months (range, 14.52–36.80 months) and 36.60 months (range, 24.3–52.9 months), respectively. The median follow up was 27 months (range, 5.00–68.00 months). Radiological response and TRG were found to be a prognostic factor for OS and DFS, while tumor histology was not significantly related to survival. Conclusions Both radiological response and TRG have been shown as promising survival markers in patients treated with perioperative chemotherapy for locally advanced gastric cancer. Other predictive markers of response to chemotherapy are strongly required. PMID:29299362

Carotenoid Biosynthetic Pathways Are Regulated by a Network of Multiple Cascades of Alternative Sigma Factors in Azospirillum brasilense Sp7.

PubMed

Rai, Ashutosh Kumar; Dubey, Ashutosh Prakash; Kumar, Santosh; Dutta, Debashis; Mishra, Mukti Nath; Singh, Bhupendra Narain; Tripathi, Anil Kumar

2016-11-01

Carotenoids constitute an important component of the defense system against photooxidative stress in bacteria. In Azospirillum brasilense Sp7, a nonphotosynthetic rhizobacterium, carotenoid synthesis is controlled by a pair of extracytoplasmic function sigma factors (RpoEs) and their cognate zinc-binding anti-sigma factors (ChrRs). Its genome harbors two copies of the gene encoding geranylgeranyl pyrophosphate synthase (CrtE), the first critical step in the carotenoid biosynthetic pathway in bacteria. Inactivation of each of two crtE paralogs found in A. brasilense caused reduction in carotenoid content, suggesting their involvement in carotenoid synthesis. However, the effect of crtE1 deletion was more pronounced than that of crtE2 deletion. Out of the five paralogs of rpoH in A. brasilense, overexpression of rpoH1 and rpoH2 enhanced carotenoid synthesis. Promoters of crtE2 and rpoH2 were found to be dependent on RpoH2 and RpoE1, respectively. Using a two-plasmid system in Escherichia coli, we have shown that the crtE2 gene of A. brasilense Sp7 is regulated by two cascades of sigma factors: one consisting of RpoE1and RpoH2 and the other consisting of RpoE2 and RpoH1. In addition, expression of crtE1 was upregulated indirectly by RpoE1 and RpoE2. This study shows, for the first time in any carotenoid-producing bacterium, that the regulation of carotenoid biosynthetic pathway involves a network of multiple cascades of alternative sigma factors. Carotenoids play a very important role in coping with photooxidative stress in prokaryotes and eukaryotes. Although extracytoplasmic function (ECF) sigma factors are known to directly regulate the expression of carotenoid biosynthetic genes in bacteria, regulation of carotenoid biosynthesis by one or multiple cascades of sigma factors had not been reported. This study provides the first evidence of the involvement of multiple cascades of sigma factors in the regulation of carotenoid synthesis in any bacterium by showing the regulation of a gene encoding geranylgeranyl pyrophosphate synthase (crtE2) by RpoE1→RpoH2→CrtE2 and RpoE2→RpoH1→CrtE2 cascades in A. brasilense It also provides an insight into existence of an additional cascade or cascades regulating expression of another paralog of crtE. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Carotenoid Biosynthetic Pathways Are Regulated by a Network of Multiple Cascades of Alternative Sigma Factors in Azospirillum brasilense Sp7

PubMed Central

Rai, Ashutosh Kumar; Dubey, Ashutosh Prakash; Kumar, Santosh; Dutta, Debashis; Mishra, Mukti Nath; Singh, Bhupendra Narain

2016-01-01

ABSTRACT Carotenoids constitute an important component of the defense system against photooxidative stress in bacteria. In Azospirillum brasilense Sp7, a nonphotosynthetic rhizobacterium, carotenoid synthesis is controlled by a pair of extracytoplasmic function sigma factors (RpoEs) and their cognate zinc-binding anti-sigma factors (ChrRs). Its genome harbors two copies of the gene encoding geranylgeranyl pyrophosphate synthase (CrtE), the first critical step in the carotenoid biosynthetic pathway in bacteria. Inactivation of each of two crtE paralogs found in A. brasilense caused reduction in carotenoid content, suggesting their involvement in carotenoid synthesis. However, the effect of crtE1 deletion was more pronounced than that of crtE2 deletion. Out of the five paralogs of rpoH in A. brasilense, overexpression of rpoH1 and rpoH2 enhanced carotenoid synthesis. Promoters of crtE2 and rpoH2 were found to be dependent on RpoH2 and RpoE1, respectively. Using a two-plasmid system in Escherichia coli, we have shown that the crtE2 gene of A. brasilense Sp7 is regulated by two cascades of sigma factors: one consisting of RpoE1and RpoH2 and the other consisting of RpoE2 and RpoH1. In addition, expression of crtE1 was upregulated indirectly by RpoE1 and RpoE2. This study shows, for the first time in any carotenoid-producing bacterium, that the regulation of carotenoid biosynthetic pathway involves a network of multiple cascades of alternative sigma factors. IMPORTANCE Carotenoids play a very important role in coping with photooxidative stress in prokaryotes and eukaryotes. Although extracytoplasmic function (ECF) sigma factors are known to directly regulate the expression of carotenoid biosynthetic genes in bacteria, regulation of carotenoid biosynthesis by one or multiple cascades of sigma factors had not been reported. This study provides the first evidence of the involvement of multiple cascades of sigma factors in the regulation of carotenoid synthesis in any bacterium by showing the regulation of a gene encoding geranylgeranyl pyrophosphate synthase (crtE2) by RpoE1→RpoH2→CrtE2 and RpoE2→RpoH1→CrtE2 cascades in A. brasilense. It also provides an insight into existence of an additional cascade or cascades regulating expression of another paralog of crtE. PMID:27551017

ESO/ST-ECF Data Analysis Workshop, 5th, Garching, Germany, Apr. 26, 27, 1993, Proceedings

NASA Astrophysics Data System (ADS)

Grosbol, Preben; de Ruijsscher, Resy

1993-01-01

Various papers on astronomical data analysis are presented. Individual optics addressed include: surface photometry of early-type galaxies, wavelet transform and adaptive filtering, package for surface photometry of galaxies, calibration of large-field mosaics, surface photometry of galaxies with HST, wavefront-supported image deconvolution, seeing effects on elliptical galaxies, multiple algorithms deconvolution program, enhancement of Skylab X-ray images, MIDAS procedures for the image analysis of E-S0 galaxies, photometric data reductions under MIDAS, crowded field photometry with deconvolved images, the DENIS Deep Near Infrared Survey. Also discussed are: analysis of astronomical time series, detection of low-amplitude stellar pulsations, new SOT method for frequency analysis, chaotic attractor reconstruction and applications to variable stars, reconstructing a 1D signal from irregular samples, automatic analysis for time series with large gaps, prospects for content-based image retrieval, redshift survey in the South Galactic Pole Region.

The AREST CF experience in biobanking - More than just tissues, tubes and time.

PubMed

Garratt, Luke W; Kicic, Anthony; Robertson, Colin; Ranganathan, Sarath; Sly, Peter D; Stick, Stephen M

2017-09-01

Research to further improve outcomes for people with CF is dependent upon well characterised, archived and accessible clinical specimens. The recent article by Beekman et al. published in Journal of Cystic Fibrosis summarised a scientific meeting at the 13th ECFS Basic Science Conference. This meeting discussed how well-annotated, clinical biobanks for CF could be established in Europe to meet the needs of therapeutic development. The Australian Respiratory Early Surveillance Team for Cystic Fibrosis (AREST CF) has conducted biobanking of CF research and clinical specimens since the late 1990s and is custodian of the most comprehensive paediatric CF biobank in the world that focuses on the first years of life. This short communication will describe the approach undertaken by AREST CF in establishing a clinical specimen biobank. Copyright © 2017 European Cystic Fibrosis Society. Published by Elsevier B.V. All rights reserved.

Fibroblast growth factor regulates insulin-like growth factor-binding protein production by vascular smooth muscle cells.

PubMed

Ververis, J; Ku, L; Delafontaine, P

1994-02-01

Insulin-like growth factor I is an important mitogen for vascular smooth muscle cells, and its effects are regulated by several binding proteins. Western ligand blotting of conditioned medium from rat aortic smooth muscle cells detected a 24 kDa binding protein and a 28 kDa glycosylated variant of this protein, consistent with insulin-like growth factor binding protein-4 by size. Low amounts of a glycosylated 38 to 42 kDa doublet (consistent with binding protein-3) and a 31 kDa non-glycosylated protein also were present. Basic fibroblast growth factor markedly increased secretion of the 24 kDa binding protein and its 28 kDa glycosylated variant. This effect was dose- and time-dependent and was inhibited by co-incubation with cycloheximide. Crosslinking of [125I]-insulin-like growth factor I to cell monolayers revealed no surface-associated binding proteins, either basally or after agonist treatment. Induction of binding protein production by fibroblast growth factor at sites of vascular injury may be important in vascular proliferative responses in vivo.

Kelch-like ECH-associated Protein 1-dependent Nuclear Factor-E2-related Factor 2 Activation in Relation to Antioxidation Induced by Sevoflurane Preconditioning.

PubMed

Cai, Min; Tong, Li; Dong, Beibei; Hou, Wugang; Shi, Likai; Dong, Hailong

2017-03-01

The authors have reported that antioxidative effects play a crucial role in the volatile anesthetic-induced neuroprotection. Accumulated evidence shows that endogenous antioxidation could be up-regulated by nuclear factor-E2-related factor 2 through multiple pathways. However, whether nuclear factor-E2-related factor 2 activation is modulated by sevoflurane preconditioning and, if so, what is the signaling cascade underlying upstream of this activation are still unknown. Sevoflurane preconditioning in mice was performed with sevoflurane (2.5%) 1 h per day for five consecutive days. Focal cerebral ischemia/reperfusion injury was induced by middle cerebral artery occlusion. Expression of nuclear factor-E2-related factor 2, kelch-like ECH-associated protein 1, manganese superoxide dismutase, thioredoxin-1, and nicotinamide adenine dinucleotide phosphate quinolone oxidoreductase-1 was detected (n = 6). The antioxidant activities and oxidative product expression were also examined. To determine the role of kelch-like ECH-associated protein 1 inhibition-dependent nuclear factor-E2-related factor 2 activation in sevoflurane preconditioning-induced neuroprotection, the kelch-like ECH-associated protein 1-nuclear factor-E2-related factor 2 signal was modulated by nuclear factor-E2-related factor 2 knockout, kelch-like ECH-associated protein 1 overexpression lentivirus, and kelch-like ECH-associated protein 1 deficiency small interfering RNA (n = 8). The infarct volume, neurologic scores, and cellular apoptosis were assessed. Sevoflurane preconditioning elicited neuroprotection and increased nuclear factor-E2-related factor 2 nuclear translocation, which in turn up-regulated endogenous antioxidation and reduced oxidative injury. Sevoflurane preconditioning reduced kelch-like ECH-associated protein 1 expression. Nuclear factor-E2-related factor 2 ablation abolished neuroprotection and reversed sevoflurane preconditioning by mediating the up-regulation of antioxidants. Kelch-like ECH-associated protein 1 overexpression reversed nuclear factor-E2-related factor 2 up-regulation and abolished the neuroprotection induced by sevoflurane preconditioning. Kelch-like ECH-associated protein 1 small interfering RNA administration improved nuclear factor-E2-related factor 2 expression and the outcome of mice subjected to ischemia/reperfusion injury. Kelch-like ECH-associated protein 1 down-regulation-dependent nuclear factor-E2-related factor 2 activation underlies the ability of sevoflurane preconditioning to activate the endogenous antioxidant response, which elicits its neuroprotection.

Protein-protein interactions in the regulation of WRKY transcription factors.

PubMed

Chi, Yingjun; Yang, Yan; Zhou, Yuan; Zhou, Jie; Fan, Baofang; Yu, Jing-Quan; Chen, Zhixiang

2013-03-01

It has been almost 20 years since the first report of a WRKY transcription factor, SPF1, from sweet potato. Great progress has been made since then in establishing the diverse biological roles of WRKY transcription factors in plant growth, development, and responses to biotic and abiotic stress. Despite the functional diversity, almost all analyzed WRKY proteins recognize the TTGACC/T W-box sequences and, therefore, mechanisms other than mere recognition of the core W-box promoter elements are necessary to achieve the regulatory specificity of WRKY transcription factors. Research over the past several years has revealed that WRKY transcription factors physically interact with a wide range of proteins with roles in signaling, transcription, and chromatin remodeling. Studies of WRKY-interacting proteins have provided important insights into the regulation and mode of action of members of the important family of transcription factors. It has also emerged that the slightly varied WRKY domains and other protein motifs conserved within each of the seven WRKY subfamilies participate in protein-protein interactions and mediate complex functional interactions between WRKY proteins and between WRKY and other regulatory proteins in the modulation of important biological processes. In this review, we summarize studies of protein-protein interactions for WRKY transcription factors and discuss how the interacting partners contribute, at different levels, to the establishment of the complex regulatory and functional network of WRKY transcription factors.

Factor VII and protein C are phosphatidic acid-binding proteins.

PubMed

Tavoosi, Narjes; Smith, Stephanie A; Davis-Harrison, Rebecca L; Morrissey, James H

2013-08-20

Seven proteins in the human blood clotting cascade bind, via their GLA (γ-carboxyglutamate-rich) domains, to membranes containing exposed phosphatidylserine (PS), although with membrane binding affinities that vary by 3 orders of magnitude. Here we employed nanodiscs of defined phospholipid composition to quantify the phospholipid binding specificities of these seven clotting proteins. All bound preferentially to nanobilayers in which PS headgroups contained l-serine versus d-serine. Surprisingly, however, nanobilayers containing phosphatidic acid (PA) bound substantially more of two of these proteins, factor VIIa and activated protein C, than did equivalent bilayers containing PS. Consistent with this finding, liposomes containing PA supported higher proteolytic activity by factor VIIa and activated protein C toward their natural substrates (factors X and Va, respectively) than did PS-containing liposomes. Moreover, treating activated human platelets with phospholipase D enhanced the rates of factor X activation by factor VIIa in the presence of soluble tissue factor. We hypothesize that factor VII and protein C bind preferentially to the monoester phosphate of PA because of its accessibility and higher negative charge compared with the diester phosphates of most other phospholipids. We further found that phosphatidylinositol 4-phosphate, which contains a monoester phosphate attached to its myo-inositol headgroup, also supported enhanced enzymatic activity of factor VIIa and activated protein C. We conclude that factor VII and protein C bind preferentially to monoester phosphates, which may have implications for the function of these proteases in vivo.

Analyses of mitochondrial genes reveal two sympatric but genetically divergent lineages of Rhipicephalus appendiculatus in Kenya.

PubMed

Kanduma, Esther G; Mwacharo, Joram M; Githaka, Naftaly W; Kinyanjui, Peter W; Njuguna, Joyce N; Kamau, Lucy M; Kariuki, Edward; Mwaura, Stephen; Skilton, Robert A; Bishop, Richard P

2016-06-22

The ixodid tick Rhipicephalus appendiculatus transmits the apicomplexan protozoan parasite Theileria parva, which causes East coast fever (ECF), the most economically important cattle disease in eastern and southern Africa. Recent analysis of micro- and minisatellite markers showed an absence of geographical and host-associated genetic sub-structuring amongst field populations of R. appendiculatus in Kenya. To assess further the phylogenetic relationships between field and laboratory R. appendiculatus tick isolates, this study examined sequence variations at two mitochondrial genes, cytochrome c oxidase subunit I (COI) and 12S ribosomal RNA (rRNA), and the nuclear encoded ribosomal internal transcribed spacer 2 (ITS2) of the rRNA gene, respectively. The analysis of 332 COI sequences revealed 30 polymorphic sites, which defined 28 haplotypes that were separated into two distinct haplogroups (A and B). Inclusion of previously published haplotypes in our analysis revealed a high degree of phylogenetic complexity never reported before in haplogroup A. Neither haplogroup however, showed any clustering pattern related to either the geographical sampling location, the type of tick sampled (laboratory stocks vs field populations) or the mammalian host species. This finding was supported by the results obtained from the analysis of 12S rDNA sequences. Analysis of molecular variance (AMOVA) indicated that 90.8 % of the total genetic variation was explained by the two haplogroups, providing further support for their genetic divergence. These results were, however, not replicated by the nuclear transcribed ITS2 sequences likely because of recombination between the nuclear genomes maintaining a high level of genetic sequence conservation. COI and 12S rDNA are better markers than ITS2 for studying intraspecific diversity. Based on these genes, two major genetic groups of R. appendiculatus that have gone through a demographic expansion exist in Kenya. The two groups show no phylogeographic structure or correlation with the type of host species from which the ticks were collected, nor to the evolutionary and breeding history of the species. The two lineages may have a wide geographic distribution range in eastern and southern Africa. The findings of this study may have implications for the spread and control of R. appendiculatus, and indirectly, on the transmission dynamics of ECF.

Estimates of effective population size and inbreeding in South African indigenous chicken populations: implications for the conservation of unique genetic resources.

PubMed

Mtileni, Bohani; Dzama, Kennedy; Nephawe, Khathutshelo; Rhode, Clint

2016-06-01

Conservation of locally adapted indigenous livestock breeds has become an important objective in sustainable animal breeding, as these breeds represent a unique genetic resource. Therefore, the Agricultural Research Council of South Africa initiated a conservation programme for four South African indigenous chicken breeds. The evaluation and monitoring of the genetic constitution of these conservation flocks is important for proper management of the conservation programme. Using molecular genetic analyses, the effective population sizes and relatedness of these conservation flocks were compared to village (field) chicken populations from which they were derived. Genetic diversity within and between these populations are further discussed within the context of population size. The conservation flocks for the respective breeds had relatively small effective population sizes (point estimate range 38.6-78.6) in comparison to the field populations (point estimate range 118.9-580.0). Furthermore, evidence supports a transient heterozygous excess, generally associated with the occurrence of a recent population bottleneck. Genetic diversity, as measured by the number of alleles, heterozygosity and information index, was also significantly reduced in the conservation flocks. The average relatedness amongst the conservation flocks was high, whilst it remained low for the field populations. There was also significant evidence for population differentiation between field and conservation populations. F st estimates for conservation flocks were moderate to high with a maximum reached between VD_C and VD_F (0.285). However, F st estimates for field population were excessively low between the NN_C and EC_F (0.007) and between EC_F and OV_F (0.009). The significant population differentiation of the conservation flocks from their geographically correlated field populations of origin is further supported by the analysis of molecular variance (AMOVA), with 10.51 % of genetic diversity ascribed to population differences within groups (F SC = 0.106). The results suggest that significant genetic erosion has occurred within the conservation flocks due to inbreeding, pronounced effects of random drift and selection. It might be necessary to introduce new breeding individuals from the respective field populations in order to increase the effective population sizes of the conservation flocks and counter the effects of genetic erosion.

Role of carbohydrate in multimeric structure of factor VIII/von Willebrand factor protein.

PubMed Central

Gralnick, H R; Williams, S B; Rick, M E

1983-01-01

The carbohydrate moiety of the factor VIII/von Willebrand (vW) factor protein is important in the expression of vW factor activity and the intravascular survival of the protein. Studies of normal human factor VIII/vW factor protein indicate that there is a requirement of a full complement of penultimate galactose for the maintenance of a normal multimeric structure. Release of penultimate galactose by beta-galactosidase or modification by galactose oxidase results in loss of the largest molecular weight multimers and increased numbers of intermediate and smaller multimers. In contrast, terminal galactose on the factor VIII/vW factor protein does not appear to play a significant role in the maintenance of the multimer organization. The abnormalities in multimeric structure and molecular size were demonstrated by NaDodSO4/polyacrylamide/agarose gel electrophoresis, NaDodSO4/glyoxyl-agarose electrophoresis, and sucrose density ultracentrifugation. These studies indicate that the penultimate galactose plays a role in the maintenance of the largest multimers of the factor VIII/vW factor protein. This may explain why, in some patients with variant forms of vW disease, a carbohydrate abnormality also may affect the multimeric structure of the plasma factor VIII/vW factor protein. Images PMID:6601805

Lack of A-factor production induces the expression of nutrient scavenging and stress-related proteins in Streptomyces griseus.

PubMed

Birkó, Zsuzsanna; Swiatek, Magdalena; Szájli, Emília; Medzihradszky, Katalin F; Vijgenboom, Erik; Penyige, András; Keseru, Judit; van Wezel, Gilles P; Biró, Sándor

2009-10-01

The small gamma-butyrolactone A-factor is an important autoregulatory signaling molecule for the soil-inhabiting streptomycetes. Starvation is a major trigger for development, and nutrients are provided by degradation of the vegetative mycelium via a process of programmed cell death, reusing proteins, nucleic acids, and cell wall material. The A-factor regulon includes many extracellular hydrolases. Here we show via proteomics analysis that many nutrient-scavenging and stress-related proteins were overexpressed in an A-factor non-producing mutant of Streptomyces griseus B-2682. Transcript analysis showed that this is primarily due to differential transcription of the target genes during early development. The targets include proteins relating to nutrient stress and environmental stress and an orthologue of the Bacillus sporulation control protein Spo0M. The enhanced expression of these proteins underlines the stress that is generated by the absence of A-factor. Wild-type developmental gene expression was restored to the A-factor non-producing mutant by the signaling protein Factor C in line with our earlier observation that Factor C triggers A-factor production.

Extreme climatic conditions and health service utilisation across rural and metropolitan New South Wales

NASA Astrophysics Data System (ADS)

Jegasothy, Edward; McGuire, Rhydwyn; Nairn, John; Fawcett, Robert; Scalley, Benjamin

2017-08-01

Periods of successive extreme heat and cold temperature have major effects on human health and increase rates of health service utilisation. The severity of these events varies between geographic locations and populations. This study aimed to estimate the effects of heat waves and cold waves on health service utilisation across urban, regional and remote areas in New South Wales (NSW), Australia, during the 10-year study period 2005-2015. We divided the state into three regions and used 24 over-dispersed or zero-inflated Poisson time-series regression models to estimate the effect of heat waves and cold waves, of three levels of severity, on the rates of ambulance call-outs, emergency department (ED) presentations and mortality. We defined heat waves and cold waves using excess heat factor (EHF) and excess cold factor (ECF) metrics, respectively. Heat waves generally resulted in increased rates of ambulance call-outs, ED presentations and mortality across the three regions and the entire state. For all of NSW, very intense heat waves resulted in an increase of 10.8% (95% confidence interval (CI) 4.5, 17.4%) in mortality, 3.4% (95% CI 0.8, 7.8%) in ED presentations and 10.9% (95% CI 7.7, 14.2%) in ambulance call-outs. Cold waves were shown to have significant effects on ED presentations (9.3% increase for intense events, 95% CI 8.0-10.6%) and mortality (8.8% increase for intense events, 95% CI 2.1-15.9%) in outer regional and remote areas. There was little evidence for an effect from cold waves on health service utilisation in major cities and inner regional areas. Heat waves have a large impact on health service utilisation in NSW in both urban and rural settings. Cold waves also have significant effects in outer regional and remote areas. EHF is a good predictor of health service utilisation for heat waves, although service needs may differ between urban and rural areas.

Nitrogen-to-Protein Conversion Factors for Three Edible Insects: Tenebrio molitor, Alphitobius diaperinus, and Hermetia illucens.

PubMed

Janssen, Renske H; Vincken, Jean-Paul; van den Broek, Lambertus A M; Fogliano, Vincenzo; Lakemond, Catriona M M

2017-03-22

Insects are considered a nutritionally valuable source of alternative proteins, and their efficient protein extraction is a prerequisite for large-scale use. The protein content is usually calculated from total nitrogen using the nitrogen-to-protein conversion factor (Kp) of 6.25. This factor overestimates the protein content, due to the presence of nonprotein nitrogen in insects. In this paper, a specific Kp of 4.76 ± 0.09 was calculated for larvae from Tenebrio molitor, Alphitobius diaperinus, and Hermetia illucens, using amino acid analysis. After protein extraction and purification, a Kp factor of 5.60 ± 0.39 was found for the larvae of three insect species studied. We propose to adopt these Kp values for determining protein content of insects to avoid overestimation of the protein content.

Nitrogen-to-Protein Conversion Factors for Three Edible Insects: Tenebrio molitor, Alphitobius diaperinus, and Hermetia illucens

PubMed Central

2017-01-01

Insects are considered a nutritionally valuable source of alternative proteins, and their efficient protein extraction is a prerequisite for large-scale use. The protein content is usually calculated from total nitrogen using the nitrogen-to-protein conversion factor (Kp) of 6.25. This factor overestimates the protein content, due to the presence of nonprotein nitrogen in insects. In this paper, a specific Kp of 4.76 ± 0.09 was calculated for larvae from Tenebrio molitor, Alphitobius diaperinus, and Hermetia illucens, using amino acid analysis. After protein extraction and purification, a Kp factor of 5.60 ± 0.39 was found for the larvae of three insect species studied. We propose to adopt these Kp values for determining protein content of insects to avoid overestimation of the protein content. PMID:28252948

Fusion proteins comprising annexin V and Kunitz protease inhibitors are highly potent thrombogenic site-directed anticoagulants

PubMed Central

Chen, Hsiu-Hui; Vicente, Cristina P.; He, Li; Tollefsen, Douglas M.; Wun, Tze-Chein

2005-01-01

The anionic phospholipid, phosphatidyl-l-serine (PS), is sequestered in the inner layer of the plasma membrane in normal cells. Upon injury, activation, and apoptosis, PS becomes exposed on the surfaces of cells and sheds microparticles, which are procoagulant. Coagulation is initiated by formation of a tissue factor/factor VIIa complex on PS-exposed membranes and propagated through the assembly of intrinsic tenase (factor VIIIa/factor IXa), prothrombinase (factor Va/factor Xa), and factor XIa complexes on PS-exposed activated platelets. We constructed a novel series of recombinant anticoagulant fusion proteins by linking annexin V (ANV), a PS-binding protein, to the Kunitz-type protease inhibitor (KPI) domain of tick anticoagulant protein, an aprotinin mutant (6L15), amyloid β-protein precursor, or tissue factor pathway inhibitor. The resulting ANV-KPI fusion proteins were 6- to 86-fold more active than recombinant tissue factor pathway inhibitor and tick anticoagulant protein in an in vitro tissue factor–initiated clotting assay. The in vivo antithrombotic activities of the most active constructs were 3- to 10-fold higher than that of ANV in a mouse arterial thrombosis model. ANV-KPI fusion proteins represent a new class of anticoagulants that specifically target the anionic membrane-associated coagulation enzyme complexes present at sites of thrombogenesis and are potentially useful as antithrombotic agents. PMID:15677561

Shotgun Proteomic Analysis of Plasma from Dairy Cattle Suffering from Footrot: Characterization of Potential Disease-Associated Factors

PubMed Central

Sun, Dongbo; Zhang, Hong; Guo, Donghua; Sun, Anguo; Wang, Hongbin

2013-01-01

The plasma proteome of healthy dairy cattle and those with footrot was investigated using a shotgun LC-MS/MS approach. In total, 648 proteins were identified in healthy plasma samples, of which 234 were non-redundant proteins and 123 were high-confidence proteins; 712 proteins were identified from footrot plasma samples, of which 272 were non-redundant proteins and 138 were high-confidence proteins. The high-confidence proteins showed significant differences between healthy and footrot plasma samples in molecular weight, isoelectric points and the Gene Ontology categories. 22 proteins were found that may differentiate between the two sets of plasma proteins, of which 16 potential differential expression (PDE) proteins from footrot plasma involved in immunoglobulins, innate immune recognition molecules, acute phase proteins, regulatory proteins, and cell adhesion and cytoskeletal proteins; 6 PDE proteins from healthy plasma involved in regulatory proteins, cytoskeletal proteins and coagulation factors. Of these PDE proteins, haptoglobin, SERPINA10 protein, afamin precursor, haptoglobin precursor, apolipoprotein D, predicted peptidoglycan recognition protein L (PGRP-L) and keratan sulfate proteoglycan (KS-PG) were suggested to be potential footrot-associated factors. The PDE proteins PGRP-L and KS-PG were highlighted as potential biomarkers of footrot in cattle. The resulting protein lists and potential differentially expressed proteins may provide valuable information to increase understanding of plasma protein profiles in cattle and to assist studies of footrot-associated factors. PMID:23418487

Complement factor H family proteins in their non-canonical role as modulators of cellular functions.

PubMed

Józsi, Mihály; Schneider, Andrea E; Kárpáti, Éva; Sándor, Noémi

2018-01-04

Complement factor H is a major regulator of the alternative pathway of the complement system. The factor H-related proteins are less characterized, but recent data indicate that they rather promote complement activation. These proteins have some common ligands with factor H and have both overlapping and distinct functions depending on domain composition and the degree of conservation of amino acid sequence. Factor H and some of the factor H-related proteins also appear in a non-canonical function that is beyond their role in the modulation of complement activation. This review covers our current understanding on this emerging role of factor H family proteins in modulating the activation and function of various cells by binding to receptors or receptor ligands. Copyright © 2018 Elsevier Ltd. All rights reserved.

Multiscale weighted colored graphs for protein flexibility and rigidity analysis

NASA Astrophysics Data System (ADS)

Bramer, David; Wei, Guo-Wei

2018-02-01

Protein structural fluctuation, measured by Debye-Waller factors or B-factors, is known to correlate to protein flexibility and function. A variety of methods has been developed for protein Debye-Waller factor prediction and related applications to domain separation, docking pose ranking, entropy calculation, hinge detection, stability analysis, etc. Nevertheless, none of the current methodologies are able to deliver an accuracy of 0.7 in terms of the Pearson correlation coefficients averaged over a large set of proteins. In this work, we introduce a paradigm-shifting geometric graph model, multiscale weighted colored graph (MWCG), to provide a new generation of computational algorithms to significantly change the current status of protein structural fluctuation analysis. Our MWCG model divides a protein graph into multiple subgraphs based on interaction types between graph nodes and represents the protein rigidity by generalized centralities of subgraphs. MWCGs not only predict the B-factors of protein residues but also accurately analyze the flexibility of all atoms in a protein. The MWCG model is validated over a number of protein test sets and compared with many standard methods. An extensive numerical study indicates that the proposed MWCG offers an accuracy of over 0.8 and thus provides perhaps the first reliable method for estimating protein flexibility and B-factors. It also simultaneously predicts all-atom flexibility in a molecule.

Human Cells as Platform to Produce Gamma-Carboxylated Proteins.

PubMed

de Sousa Bomfim, Aline; de Freitas, Marcela Cristina Corrêa; Covas, Dimas Tadeu; de Sousa Russo, Elisa Maria

2018-01-01

The gamma-carboxylated proteins belong to a family of proteins that depend on vitamin K for normal biosynthesis. The major representative gamma-carboxylated proteins are the coagulation system proteins, for example, factor VII, factor IX, factor X, prothrombin, and proteins C, S, and Z. These molecules have harbored posttranslational modifications, such as glycosylation and gamma-carboxylation, and for this reason they need to be produced in mammalian cell lines. Human cells lines have emerged as the most promising alternative to the production of gamma-carboxylated proteins. In this chapter, the methods to generate human cells as a platform to produce gamma-carboxylated proteins, for example the coagulation factors VII and IX, are presented. From the cell line modification up to the vitamin K adaptation of the produced cells is described in the protocols presented in this chapter.

Protein-Protein Interactions within Late Pre-40S Ribosomes

PubMed Central

Campbell, Melody G.; Karbstein, Katrin

2011-01-01

Ribosome assembly in eukaryotic organisms requires more than 200 assembly factors to facilitate and coordinate rRNA transcription, processing, and folding with the binding of the ribosomal proteins. Many of these assembly factors bind and dissociate at defined times giving rise to discrete assembly intermediates, some of which have been partially characterized with regards to their protein and RNA composition. Here, we have analyzed the protein-protein interactions between the seven assembly factors bound to late cytoplasmic pre-40S ribosomes using recombinant proteins in binding assays. Our data show that these factors form two modules: one comprising Enp1 and the export adaptor Ltv1 near the beak structure, and the second comprising the kinase Rio2, the nuclease Nob1, and a regulatory RNA binding protein Dim2/Pno1 on the front of the head. The GTPase-like Tsr1 and the universally conserved methylase Dim1 are also peripherally connected to this second module. Additionally, in an effort to further define the locations for these essential proteins, we have analyzed the interactions between these assembly factors and six ribosomal proteins: Rps0, Rps3, Rps5, Rps14, Rps15 and Rps29. Together, these results and previous RNA-protein crosslinking data allow us to propose a model for the binding sites of these seven assembly factors. Furthermore, our data show that the essential kinase Rio2 is located at the center of the pre-ribosomal particle and interacts, directly or indirectly, with every other assembly factor, as well as three ribosomal proteins required for cytoplasmic 40S maturation. These data suggest that Rio2 could play a central role in regulating cytoplasmic maturation steps. PMID:21283762

Protein-protein interactions within late pre-40S ribosomes.

PubMed

Campbell, Melody G; Karbstein, Katrin

2011-01-20

Ribosome assembly in eukaryotic organisms requires more than 200 assembly factors to facilitate and coordinate rRNA transcription, processing, and folding with the binding of the ribosomal proteins. Many of these assembly factors bind and dissociate at defined times giving rise to discrete assembly intermediates, some of which have been partially characterized with regards to their protein and RNA composition. Here, we have analyzed the protein-protein interactions between the seven assembly factors bound to late cytoplasmic pre-40S ribosomes using recombinant proteins in binding assays. Our data show that these factors form two modules: one comprising Enp1 and the export adaptor Ltv1 near the beak structure, and the second comprising the kinase Rio2, the nuclease Nob1, and a regulatory RNA binding protein Dim2/Pno1 on the front of the head. The GTPase-like Tsr1 and the universally conserved methylase Dim1 are also peripherally connected to this second module. Additionally, in an effort to further define the locations for these essential proteins, we have analyzed the interactions between these assembly factors and six ribosomal proteins: Rps0, Rps3, Rps5, Rps14, Rps15 and Rps29. Together, these results and previous RNA-protein crosslinking data allow us to propose a model for the binding sites of these seven assembly factors. Furthermore, our data show that the essential kinase Rio2 is located at the center of the pre-ribosomal particle and interacts, directly or indirectly, with every other assembly factor, as well as three ribosomal proteins required for cytoplasmic 40S maturation. These data suggest that Rio2 could play a central role in regulating cytoplasmic maturation steps.

Stable expression of recombinant human coagulation factor XIII in protein-free suspension culture of Chinese hamster ovary cells.

PubMed

Chun, B H; Bang, W G; Park, Y K; Woo, S K

2001-11-01

The recombinant a and bsubunits for human coagulation factor XIII were transfected into Chinese hamster ovary (CHO) cells. CHO cells were amplified and selected with methotrexate in adherent cultures containing serum, and CHO 1-62 cells were later selected in protein-free medium. To develop a recombinant factor XIII production process in a suspension culture, we have investigated the growth characteristics of CHO cells and the maintenance of factor XIII expression in the culture medium. Suspension adaptation of CHO cells was performed in protein-free medium, GC-CHO-PI, by two methods, such as serum weaning and direct switching from serum containing media to protein-free media. Although the growth of CHO cells in suspension culture was affected initially by serum depletion, cell specific productivity of factor XIII showed only minor changes by the direct switching to protein-free medium during a suspension culture. As for the long-term stability of factor XIII, CHO 1-62 cells showed a stable expression of factor XIII in protein-free condition for 1000 h. These results indicate that the CHO 1-62cells can be adapted to express recombinant human factor XIII in a stable maimer in suspension culture using a protein-free medium. Our results demonstrate that enhanced cell growth in a continuous manner is achievable for factor XIII production in a protein-free medium when a perfusion bioreactor culture system with a spin filter is employed.

Using host-pathogen protein interactions to identify and characterize Francisella tularensis virulence factors.

PubMed

Wallqvist, Anders; Memišević, Vesna; Zavaljevski, Nela; Pieper, Rembert; Rajagopala, Seesandra V; Kwon, Keehwan; Yu, Chenggang; Hoover, Timothy A; Reifman, Jaques

2015-12-29

Francisella tularensis is a select bio-threat agent and one of the most virulent intracellular pathogens known, requiring just a few organisms to establish an infection. Although several virulence factors are known, we lack an understanding of virulence factors that act through host-pathogen protein interactions to promote infection. To address these issues in the highly infectious F. tularensis subsp. tularensis Schu S4 strain, we deployed a combined in silico, in vitro, and in vivo analysis to identify virulence factors and their interactions with host proteins to characterize bacterial infection mechanisms. We initially used comparative genomics and literature to identify and select a set of 49 putative and known virulence factors for analysis. Each protein was then subjected to proteome-scale yeast two-hybrid (Y2H) screens with human and murine cDNA libraries to identify potential host-pathogen protein-protein interactions. Based on the bacterial protein interaction profile with both hosts, we selected seven novel putative virulence factors for mutant construction and animal validation experiments. We were able to create five transposon insertion mutants and used them in an intranasal BALB/c mouse challenge model to establish 50 % lethal dose estimates. Three of these, ΔFTT0482c, ΔFTT1538c, and ΔFTT1597, showed attenuation in lethality and can thus be considered novel F. tularensis virulence factors. The analysis of the accompanying Y2H data identified intracellular protein trafficking between the early endosome to the late endosome as an important component in virulence attenuation for these virulence factors. Furthermore, we also used the Y2H data to investigate host protein binding of two known virulence factors, showing that direct protein binding was a component in the modulation of the inflammatory response via activation of mitogen-activated protein kinases and in the oxidative stress response. Direct interactions with specific host proteins and the ability to influence interactions among host proteins are important components for F. tularensis to avoid host-cell defense mechanisms and successfully establish an infection. Although direct host-pathogen protein-protein binding is only one aspect of Francisella virulence, it is a critical component in directly manipulating and interfering with cellular processes in the host cell.

Binding Mode Analysis of Zerumbone to Key Signal Proteins in the Tumor Necrosis Factor Pathway

PubMed Central

Fatima, Ayesha; Abdul, Ahmad Bustamam Hj.; Abdullah, Rasedee; Karjiban, Roghayeh Abedi; Lee, Vannajan Sanghiran

2015-01-01

Breast cancer is the second most common cancer among women worldwide. Several signaling pathways have been implicated as causative and progression agents. The tumor necrosis factor (TNF) α protein plays a dual role in promoting and inhibiting cancer depending largely on the pathway initiated by the binding of the protein to its receptor. Zerumbone, an active constituent of Zingiber zerumbet, Smith, is known to act on the tumor necrosis factor pathway upregulating tumour necrosis factor related apoptosis inducing ligand (TRAIL) death receptors and inducing apoptosis in cancer cells. Zerumbone is a sesquiterpene that is able to penetrate into the hydrophobic pockets of proteins to exert its inhibiting activity with several proteins. We found a good binding with the tumor necrosis factor, kinase κB (IKKβ) and the Nuclear factor κB (NF-κB) component proteins along the TNF pathway. Our results suggest that zerumbone can exert its apoptotic activities by inhibiting the cytoplasmic proteins. It inhibits the IKKβ kinase that activates the NF-κB and also binds to the NF-κB complex in the TNF pathway. Blocking both proteins can lead to inhibition of cell proliferating proteins to be downregulated and possibly ultimate induction of apoptosis. PMID:25629232

Effects of Non-Collagenous Proteins, TGF-β1, and PDGF-BB on Viability and Proliferation of Dental Pulp Stem Cells

PubMed Central

Tabatabaei, Fahimeh Sadat

2016-01-01

ABSTRACT Objectives The dentin matrix servers as a reservoir of growth factors, sequestered during dentinogenesis. The aim of this study was to assess the viability and proliferation of dental pulp stem cells in the presence of dentin matrix-derived non-collagenous proteins and two growth factors; platelet-derived growth factor BB and transforming growth factor beta 1. Material and Methods The dental pulp cells were isolated and cultured. The dentin proteins were extracted and purified. The MTT assay was performed for assessment of cell viability and proliferation in the presence of different concentrations of dentin proteins and growth factors during 24 - 72 h post-treatment. Results The cells treated with 250 ng/mL dentin proteins had the best viability and proliferation ability in comparison with other concentrations (P < 0.05). The MTT assay demonstrated that cells cultured with 5 ng/mL platelet-derived growth factor BB had the highest viability at each time point as compared to other groups (P < 0.05). However, in presence of platelet-derived growth factor BB alone and in combination with transforming growth factor beta 1 and dentin proteins (10 ng/mL), significant higher viability was seen at all time points (P < 0.05). The least viability and proliferation at each growth factor concentration was seen in cells treated with combination of transforming growth factor beta 1 and dentin proteins at 72 h (P < 0.05). Conclusions The results indicated that the triple combination of growth factors and matrix-derived non-collagenous proteins (especially at 10 ng/mL concentration) has mitogenic effect on dental pulp stem cells. PMID:27099698

Role of Fibroblast Growth Factor Binding Protein-1 in Mammary Development and Tumorigenesis

DTIC Science & Technology

2009-10-01

AD_________________ Award Number: W81XWH-06-1-0763 TITLE: Role of Fibroblast Growth Factor ...2009 4. TITLE AND SUBTITLE Role of Fibroblast Growth Factor Binding Protein-1 in Mammary Development 5a. CONTRACT NUMBER and Tumorigenesis...Release; Distribution Unlimited 13. SUPPLEMENTARY NOTES 14. ABSTRACT 15. SUBJECT TERMS Fibroblast Growth Factor Binding Protein-1

Effects of antinutritional factors on protein digestibility and amino acid availability in foods.

PubMed

Gilani, G Sarwar; Cockell, Kevin A; Sepehr, Estatira

2005-01-01

Digestibility of protein in traditional diets from developing countries such as India, Guatemala, and Brazil is considerably lower compared to that of protein in typical North American diets (54-78 versus 88-94%). The presence of less digestible protein fractions, high levels of insoluble fiber, and high concentrations of antinutritional factors in the diets of developing countries, which are based on less refined cereals and grain legumes as major sources of protein, are responsible for poor digestibility of protein. The effects of the presence of some of the important antinutritional factors on protein and amino digestibilities of food and feed products are reviewed in this chapter. Food and feed products may contain a number of antinutritional factors that may adversely affect protein digestibility and amino acid availability. Antinutritional factors may occur naturally, such as glucosinolates in mustard and rapeseed protein products, trypsin inhibitors and hemagglutinins in legumes, tannins in legumes and cereals, phytates in cereals and oilseeds, and gossypol in cottonseed protein products. Antinutritional factors may also be formed during heat/alkaline processing of protein products, yielding Maillard compounds, oxidized forms of sulfur amino acids, D-amino acids, and lysinoalanine (LAL, an unnatural amino acid derivative). The presence of high levels of dietary trypsin inhibitors from soybeans, kidney beans, or other grain legumes can cause substantial reductions in protein and amino acid digestibilities (up to 50%) in rats and pigs. Similarly, the presence of high levels of tannins in cereals, such as sorghum, and grain legumes, such as fababean (Vicia faba L.), can result in significantly reduced protein and amino acid digestibilities (up to 23%) in rats, poultry, and pigs. Studies involving phytase supplementation of production rations for swine or poultry have provided indirect evidence that normally encountered levels of phytates in cereals and legumes can reduce protein and amino acid digestibilities by up to 10%. D-amino acids and LAL formed during alkaline/heat treatment of proteins such as casein, lactalbumin, soy protein isolate, or wheat proteins are poorly digestible (less than 40%), and their presence can reduce protein digestibility by up to 28% in rats and pigs. A comparison of the protein digestibility determination in young (5-week) versus old (20-month) rats suggests greater susceptibility to the adverse effects of antinutritional factors in old rats than in young rats. Therefore, the inclusion of protein digestibility data obtained with young rats, as the recommended animal model, in the calculation of PDCAAS (Protein Digestibility-Corrected Amino Acid Score) may overestimate protein digestibility and quality of products, especially those containing antinutritional factors, for the elderly. For products specifically intended for the elderly, protein digestibility should be determined using more mature rats.

[Regulation of heat shock gene expression in response to stress].

PubMed

Garbuz, D G

2017-01-01

Heat shock (HS) genes, or stress genes, code for a number of proteins that collectively form the most ancient and universal stress defense system. The system determines the cell capability of adaptation to various adverse factors and performs a variety of auxiliary functions in normal physiological conditions. Common stress factors, such as higher temperatures, hypoxia, heavy metals, and others, suppress transcription and translation for the majority of genes, while HS genes are upregulated. Transcription of HS genes is controlled by transcription factors of the HS factor (HSF) family. Certain HSFs are activated on exposure to higher temperatures or other adverse factors to ensure stress-induced HS gene expression, while other HSFs are specifically activated at particular developmental stages. The regulation of the main mammalian stress-inducible factor HSF1 and Drosophila melanogaster HSF includes many components, such as a variety of early warning signals indicative of abnormal cell activity (e.g., increases in intracellular ceramide, cytosolic calcium ions, or partly denatured proteins); protein kinases, which phosphorylate HSFs at various Ser residues; acetyltransferases; and regulatory proteins, such as SUMO and HSBP1. Transcription factors other than HSFs are also involved in activating HS gene transcription; the set includes D. melanogaster GAF, mammalian Sp1 and NF-Y, and other factors. Transcription of several stress genes coding for molecular chaperones of the glucose-regulated protein (GRP) family is predominantly regulated by another stress-detecting system, which is known as the unfolded protein response (UPR) system and is activated in response to massive protein misfolding in the endoplasmic reticulum and mitochondrial matrix. A translational fine tuning of HS protein expression occurs via changing the phosphorylation status of several proteins involved in translation initiation. In addition, specific signal sequences in the 5'-UTRs of some HS protein mRNAs ensure their preferential translation in stress.

The expansion and performance of national newborn screening programmes for cystic fibrosis in Europe.

PubMed

Barben, Jürg; Castellani, Carlo; Dankert-Roelse, Jeannette; Gartner, Silvia; Kashirskaya, Nataliya; Linnane, Barry; Mayell, Sarah; Munck, Anne; Sands, Dorota; Sommerburg, Olaf; Pybus, Simon; Winters, Victoria; Southern, Kevin W

2017-03-01

Newborn screening (NBS) for cystic fibrosis (CF) is a well-established public health strategy with international standards. The aim of this study was to provide an update on NBS for CF in Europe and assess performance against the standards. Questionnaires were sent to key workers in each European country. In 2016, there were 17 national programmes, 4 countries with regional programmes and 25 countries not screening in Europe. All national programmes employed different protocols, with IRT-DNA the most common strategy. Five countries were not using DNA analysis. In addition, the processing and structure of programmes varied considerably. Most programmes were achieving the ECFS standards with respect to timeliness, but were less successful with respect to sensitivity and specificity. There has been a steady increase in national CF NBS programmes across Europe with variable strategies and outcomes that reflect the different approaches. Copyright © 2016 European Cystic Fibrosis Society. Published by Elsevier B.V. All rights reserved.

Factor H-related proteins.

PubMed

Józsi, Mihály; Meri, Seppo

2014-01-01

Factor H-related proteins (CFHRs) are plasma glycoproteins related in structure and antigenicity to each other and to the complement inhibitory protein factor H. Such proteins are found in most mammals but their number and domain composition vary. This chapter summarizes our current knowledge on the human factor H-related proteins. In contrast to factor H, they have no strong complement inhibitory activity, although for some of them regulatory or complement modulatory activity has been reported. A common feature of CFHRs is that they bind to the C3b component of complement. Novel links between CFHRs and various diseases (C3 glomerulopathies, atypical hemolytic uremic syndrome and age-related macular degeneration) have been revealed in recent years, but we are still far from understanding their biological function.

Structural Determination of Functional Domains in Early B-cell Factor (EBF) Family of Transcription Factors Reveals Similarities to Rel DNA-binding Proteins and a Novel Dimerization Motif*

PubMed Central

Siponen, Marina I.; Wisniewska, Magdalena; Lehtiö, Lari; Johansson, Ida; Svensson, Linda; Raszewski, Grzegorz; Nilsson, Lennart; Sigvardsson, Mikael; Berglund, Helena

2010-01-01

The early B-cell factor (EBF) transcription factors are central regulators of development in several organs and tissues. This protein family shows low sequence similarity to other protein families, which is why structural information for the functional domains of these proteins is crucial to understand their biochemical features. We have used a modular approach to determine the crystal structures of the structured domains in the EBF family. The DNA binding domain reveals a striking resemblance to the DNA binding domains of the Rel homology superfamily of transcription factors but contains a unique zinc binding structure, termed zinc knuckle. Further the EBF proteins contain an IPT/TIG domain and an atypical helix-loop-helix domain with a novel type of dimerization motif. The data presented here provide insights into unique structural features of the EBF proteins and open possibilities for detailed molecular investigations of this important transcription factor family. PMID:20592035

Membrane re-modelling by BAR domain superfamily proteins via molecular and non-molecular factors.

PubMed

Nishimura, Tamako; Morone, Nobuhiro; Suetsugu, Shiro

2018-04-17

Lipid membranes are structural components of cell surfaces and intracellular organelles. Alterations in lipid membrane shape are accompanied by numerous cellular functions, including endocytosis, intracellular transport, and cell migration. Proteins containing Bin-Amphiphysin-Rvs (BAR) domains (BAR proteins) are unique, because their structures correspond to the membrane curvature, that is, the shape of the lipid membrane. BAR proteins present at high concentration determine the shape of the membrane, because BAR domain oligomers function as scaffolds that mould the membrane. BAR proteins co-operate with various molecular and non-molecular factors. The molecular factors include cytoskeletal proteins such as the regulators of actin filaments and the membrane scission protein dynamin. Lipid composition, including saturated or unsaturated fatty acid tails of phospholipids, also affects the ability of BAR proteins to mould the membrane. Non-molecular factors include the external physical forces applied to the membrane, such as tension and friction. In this mini-review, we will discuss how the BAR proteins orchestrate membrane dynamics together with various molecular and non-molecular factors. © 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Enhanced functional recombinant factor VII production by HEK 293 cells stably transfected with VKORC1 where the gamma-carboxylase inhibitor calumenin is stably suppressed by shRNA transfection.

PubMed

Wajih, Nadeem; Owen, John; Wallin, Reidar

2008-01-01

Recombinant members of the vitamin K-dependent protein family (factors IX and VII and protein C) have become important pharmaceuticals in treatment of bleeding disorders and sepsis. However, because the in vivo gamma-carboxylation system in stable cell lines used for transfection has a limited capacity of post translational gamma-carboxylation, the recovery of fully gamma-carboxylated and functional proteins is low. In this work we have engineered recombinant factor VII producing HEK 293 cells to stably overexpress VKORC1, the reduced vitamin K gamma-carboxylase cofactor and in addition stably silenced the gamma-carboxylase inhibitory protein calumenin. Stable cell lines transfected with only a factor VII cDNA had a 9% production of functional recombinant factor VII. On the other hand, these recombinant factor VII producing cells when engineered to overexpress VKORC1 and having calumenin stably suppressed more than 80% by shRNA expression, produced 68% functional factor VII. The technology presented should be applicable to all vertebrae members of the vitamin K-dependent protein family and should lower the production cost of the clinically used factors VII, IX and protein C.

Total Protein Content Determination of Microalgal Biomass by Elemental Nitrogen Analysis and a Dedicated Nitrogen-to-Protein Conversion Factor

DOE Office of Scientific and Technical Information (OSTI.GOV)

Laurens, Lieve M; Olstad-Thompson, Jessica L; Templeton, David W

Accurately determining protein content is important in the valorization of algal biomass in food, feed, and fuel markets, where these values are used for component balance calculations. Conversion of elemental nitrogen to protein is a well-accepted and widely practiced method, but depends on developing an applicable nitrogen-to-protein conversion factor. The methodology reported here covers the quantitative assessment of the total nitrogen content of algal biomass and a description of the methodology that underpins the accurate de novo calculation of a dedicated nitrogen-to-protein conversion factor.

Low-protein diet supplemented with ketoacids reduces the severity of renal disease in 5/6 nephrectomized rats: a role for KLF15.

PubMed

Gao, Xiang; Huang, Lianghu; Grosjean, Fabrizio; Esposito, Vittoria; Wu, Jianxiang; Fu, Lili; Hu, Huimin; Tan, Jiangming; He, Cijian; Gray, Susan; Jain, Mukesh K; Zheng, Feng; Mei, Changlin

2011-05-01

Dietary protein restriction is an important treatment for chronic kidney disease. Herein, we tested the effect of low-protein or low-protein plus ketoacids (KA) diet in a remnant kidney model. Rats with a remnant kidney were randomized to receive normal protein diet (22%), low-protein (6%) diet (LPD), or low-protein (5%) plus KA (1%) diet for 6 months. Protein restriction prevented proteinuria, decreased blood urea nitrogen levels, and renal lesions; however, the LPD retarded growth and decreased serum albumin levels. Supplementation with KA corrected these abnormalities and provided superior renal protection compared with protein restriction alone. The levels of Kruppel-like factor-15 (KLF15), a transcription factor shown to reduce cardiac fibrosis, were decreased in remnant kidneys. Protein restriction, which increased KLF15 levels in the normal kidney, partially recovered the levels of KLF15 in remnant kidney. The expression of KLF15 in mesangial cells was repressed by oxidative stress, transforming growth factor-β, and tumor necrosis factor (TNF)-α. The suppressive effect of TNF-α on KLF15 expression was mediated by TNF receptor-1 and nuclear factor-κB. Overexpression of KLF15 in mesangial and HEK293 cells significantly decreased fibronectin and type IV collagen mRNA levels. Furthermore, KLF15 knockout mice developed glomerulosclerosis following uninephrectomy. Thus, KLF15 may be an antifibrotic factor in the kidney, and its decreased expression may contribute to the progression of kidney disease.

Thrombophilic mutations in pre-eclampsia and pregnancy-induced hypertension.

PubMed

Omar, Siti Z; Qvist, Rajes; Khaing, Si L; Muniandy, Sekaran; Bhalla, Sunil

2008-04-01

The aim of the present study was to determine the existence or prevalence of thrombophilic markers such as Factor V Leiden, prothrombin G20210A, protein S, protein C, activated protein C and anti-thrombin in pre-eclampsia and pregnancy-induced hypertensive patients. Blood samples were collected from a total number of 124 women at the maternity unit, University of Malaya Medical Center. These included 49 patients with pre-eclampsia, 63 patients with pregnancy-induced hypertension and 12 normal pregnant women. DNA was extracted from the blood samples. Factor V Leiden (Taq I) and prothrombin G20210A (Hind III) genotyping was done on polymerase chain reaction-restriction fragment length polymorphism. Anti-thrombin activity and the concentrations of protein C, protein S and activated protein C were measured using the IL Coagulation System (Hemosil). Of the 124 subjects, one pre-eclampsia patient was homozygous for Factor V Leiden mutation but prothrombin G20210A mutation was not present in any of the subjects. The subject with Factor V Leiden mutation also had a low activated protein C resistance and a low protein S concentration. Factor V Leiden mutation is present in the Asian population and may very well serve as one of the genetic factors responsible for pre-eclampsia and other adverse pregnancy outcomes.

Complex two-component signaling regulates the general stress response in Alphaproteobacteria.

PubMed

Kaczmarczyk, Andreas; Hochstrasser, Ramon; Vorholt, Julia A; Francez-Charlot, Anne

2014-12-02

The general stress response (GSR) in Alphaproteobacteria was recently shown to be controlled by a partner-switching mechanism that is triggered by phosphorylation of the response regulator PhyR. Activation of PhyR ultimately results in release of the alternative extracytoplasmic function sigma factor σ(EcfG), which redirects transcription toward the GSR. Little is known about the signal transduction pathway(s) controlling PhyR phosphorylation. Here, we identified the single-domain response regulator (SDRR) SdrG and seven histidine kinases, PakA to PakG, belonging to the HWE/HisKA2 family as positive modulators of the GSR in Sphingomonas melonis Fr1. Phenotypic analyses, epistasis experiments, and in vitro phosphorylation assays indicate that Paks directly phosphorylate PhyR and SdrG, and that SdrG acts upstream of or in concert with PhyR, modulating its activity in a nonlinear pathway. Furthermore, we found that additional SDRRs negatively affect the GSR in a way that strictly requires PhyR and SdrG. Finally, analysis of GSR activation by thermal, osmotic, and oxidative stress indicates that Paks display different degrees of redundancy and that a specific kinase can sense multiple stresses, suggesting that the GSR senses a particular condition as a combination of, rather than individual, molecular cues. This study thus establishes the alphaproteobacterial GSR as a complex and interlinked network of two-component systems, in which multiple histidine kinases converge to PhyR, the phosphorylation of which is, in addition, subject to regulation by several SDRRs. Our finding that most HWE/HisKA2 kinases contribute to the GSR in S. melonis Fr1 opens the possibility that this notion might also be true for other Alphaproteobacteria.

Target evaluation of deoxyhypusine synthase from Theileria parva the neglected animal parasite and its relationship to Plasmodium.

PubMed

Njuguna, James T; von Koschitzky, Imke; Gerhardt, Heike; Lämmerhofer, Michael; Choucry, Ali; Pink, Mario; Schmitz-Spahnke, Simone; Bakheit, Mohammed A; Strube, Christina; Kaiser, Annette

2014-08-01

East Coast fever (ECF) is a tick-borne disease caused by the parasite Theileria parva which infects cattle. In Sub-Saharan Africa it leads to enormous economic costs. After a bite of a tick, sporozoites invade the host lymphocytes and develop into schizonts. At this stage the parasite transforms host lymphocytes resulting in the clonal expansion of infected lymphocytes. Animals develop a lymphoma like disorder after infection which is rapidly fatal. Hitherto, a few drugs of the quinone type can cure the disease. However, therapy can only be successful after early diagnosis. The genera Theileria and Plasmodium, which includes the causative agent of human malaria, are closely related apicomplexan parasites. Enzymes of the hypusine pathway, a posttranslational modification in eukaryotic initiation factor EIF-5A, have shown to be druggable targets in Plasmodium. We identified the first enzyme of the hypusine pathway from T. parva, the deoxyhypusine synthase (DHS), which is located on chromosome 2 of the Muguga strain. Transcription is significantly increased in schizonts. The expressed T. parva DHS reveals an open reading frame (ORF) of 370 amino acids after expression in Escherichia coli Rosetta cells with a molecular size of 41.26 kDa and a theoretical pI of 5.26. Screening of the Malaria Box which consists of 400 active compounds resulted in a novel heterocyclic compound with a guanyl spacer which reduced the activity of T. parva DHS to 45%. In sum, the guanyl residue seems to be an important lead structure for inhibition of Theileria DHS. Currently, more different guanyl analogues from the Malaria Box are tested in inhibitor experiments to determine their efficacy. Copyright © 2014 Elsevier Ltd. All rights reserved.

Fossil and non-fossil source contributions to atmospheric carbonaceous aerosols during extreme spring grassland fires in Eastern Europe

NASA Astrophysics Data System (ADS)

Ulevicius, V.; Byčenkienė, S.; Bozzetti, C.; Vlachou, A.; Plauškaitė, K.; Mordas, G.; Dudoitis, V.; Abbaszade, G.; Remeikis, V.; Garbaras, A.; Masalaite, A.; Blees, J.; Fröhlich, R.; Dällenbach, K. R.; Canonaco, F.; Slowik, J. G.; Dommen, J.; Zimmermann, R.; Schnelle-Kreis, J.; Salazar, G. A.; Agrios, K.; Szidat, S.; El Haddad, I.; Prévôt, A. S. H.

2015-09-01

In early spring the Baltic region is frequently affected by high pollution events due to biomass burning in that area. Here we present a comprehensive study to investigate the impact of biomass/grass burning (BB) on the evolution and composition of aerosol in Preila, Lithuania, during springtime open fires. Non-refractory submicron particulate matter (NR-PM1) was measured by an Aerodyne aerosol chemical speciation monitor (ACSM) and a source apportionment with the multilinear engine (ME-2) running the positive matrix factorization (PMF) model was applied to the organic aerosol fraction to investigate the impact of biomass/grass burning. Satellite observations over regions of biomass burning activity supported the results and identification of air mass transport to the area of investigation. Sharp increases in biomass burning tracers, such as levoglucosan up to 683 ng m-3 and black carbon (BC) up to 17 μg m-3 were observed during this period. A further separation between fossil and non-fossil primary and secondary contributions was obtained by coupling ACSM PMF results and radiocarbon (14C) measurements of the elemental (EC) and organic (OC) carbon fractions. Non-fossil organic carbon (OCnf) was the dominant fraction of PM1, with the primary (POCnf) and secondary (SOCnf) fractions contributing 26-44 % and 13-23 % to the TC, respectively. 5-8 % of the TC had a primary fossil origin (POCf), whereas the contribution of fossil secondary organic carbon (SOCf) was 4-13 %. Non-fossil EC (ECnf) and fossil EC (ECf) ranged from 13-24 % and 7-12 %, respectively. Isotope ratio of stable carbon and nitrogen isotopes were used to distinguish aerosol particles associated with solid and liquid fossil fuel burning.

Fossil and non-fossil source contributions to atmospheric carbonaceous aerosols during extreme spring grassland fires in Eastern Europe

NASA Astrophysics Data System (ADS)

Ulevicius, Vidmantas; Byčenkienė, Steigvilė; Bozzetti, Carlo; Vlachou, Athanasia; Plauškaitė, Kristina; Mordas, Genrik; Dudoitis, Vadimas; Abbaszade, Gülcin; Remeikis, Vidmantas; Garbaras, Andrius; Masalaite, Agne; Blees, Jan; Fröhlich, Roman; Dällenbach, Kaspar R.; Canonaco, Francesco; Slowik, Jay G.; Dommen, Josef; Zimmermann, Ralf; Schnelle-Kreis, Jürgen; Salazar, Gary A.; Agrios, Konstantinos; Szidat, Sönke; El Haddad, Imad; Prévôt, André S. H.

2016-05-01

In early spring the Baltic region is frequently affected by high-pollution events due to biomass burning in that area. Here we present a comprehensive study to investigate the impact of biomass/grass burning (BB) on the evolution and composition of aerosol in Preila, Lithuania, during springtime open fires. Non-refractory submicron particulate matter (NR-PM1) was measured by an Aerodyne aerosol chemical speciation monitor (ACSM) and a source apportionment with the multilinear engine (ME-2) running the positive matrix factorization (PMF) model was applied to the organic aerosol fraction to investigate the impact of biomass/grass burning. Satellite observations over regions of biomass burning activity supported the results and identification of air mass transport to the area of investigation. Sharp increases in biomass burning tracers, such as levoglucosan up to 683 ng m-3 and black carbon (BC) up to 17 µg m-3 were observed during this period. A further separation between fossil and non-fossil primary and secondary contributions was obtained by coupling ACSM PMF results and radiocarbon (14C) measurements of the elemental (EC) and organic (OC) carbon fractions. Non-fossil organic carbon (OCnf) was the dominant fraction of PM1, with the primary (POCnf) and secondary (SOCnf) fractions contributing 26-44 % and 13-23 % to the total carbon (TC), respectively. 5-8 % of the TC had a primary fossil origin (POCf), whereas the contribution of fossil secondary organic carbon (SOCf) was 4-13 %. Non-fossil EC (ECnf) and fossil EC (ECf) ranged from 13-24 and 7-13 %, respectively. Isotope ratios of stable carbon and nitrogen isotopes were used to distinguish aerosol particles associated with solid and liquid fossil fuel burning.

Industrial production of clotting factors: Challenges of expression, and choice of host cells.

PubMed

Kumar, Sampath R

2015-07-01

The development of recombinant forms of blood coagulation factors as safer alternatives to plasma derived factors marked a major advance in the treatment of common coagulation disorders. These are complex proteins, mostly enzymes or co-enzymes, involving multiple post-translational modifications, and therefore are difficult to express. This article reviews the nature of the expression challenges for the industrial production of these factors, vis-à-vis the translational and post-translational bottlenecks, as well as the choice of host cell lines for high-fidelity production. For achieving high productivities of vitamin K dependent proteins, which include factors II (prothrombin), VII, IX and X, and protein C, host cell limitation of γ-glutamyl carboxylation is a major bottleneck. Despite progress in addressing this, involvement of yet unidentified protein(s) impedes a complete cell engineering solution. Human factor VIII expresses at very low levels due to limitations at several steps in the protein secretion pathway. Protein and cell engineering, vector improvement and alternate host cells promise improvement in the productivity. Production of Von Willebrand factor is constrained by its large size, complex structure, and the need for extensive glycosylation and disulfide-bonded oligomerization. All the licensed therapeutic factors are produced in CHO, BHK or HEK293 cells. While HEK293 is a recent adoption, BHK cells appear to be disfavored. Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Position Matters: Network Centrality Considerably Impacts Rates of Protein Evolution in the Human Protein-Protein Interaction Network.

PubMed

Alvarez-Ponce, David; Feyertag, Felix; Chakraborty, Sandip

2017-06-01

The proteins of any organism evolve at disparate rates. A long list of factors affecting rates of protein evolution have been identified. However, the relative importance of each factor in determining rates of protein evolution remains unresolved. The prevailing view is that evolutionary rates are dominantly determined by gene expression, and that other factors such as network centrality have only a marginal effect, if any. However, this view is largely based on analyses in yeasts, and accurately measuring the importance of the determinants of rates of protein evolution is complicated by the fact that the different factors are often correlated with each other, and by the relatively poor quality of available functional genomics data sets. Here, we use correlation, partial correlation and principal component regression analyses to measure the contributions of several factors to the variability of the rates of evolution of human proteins. For this purpose, we analyzed the entire human protein-protein interaction data set and the human signal transduction network-a network data set of exceptionally high quality, obtained by manual curation, which is expected to be virtually free from false positives. In contrast with the prevailing view, we observe that network centrality (measured as the number of physical and nonphysical interactions, betweenness, and closeness) has a considerable impact on rates of protein evolution. Surprisingly, the impact of centrality on rates of protein evolution seems to be comparable, or even superior according to some analyses, to that of gene expression. Our observations seem to be independent of potentially confounding factors and from the limitations (biases and errors) of interactomic data sets. © The Author 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

Developmental regulation of collagenase-3 mRNA in normal, differentiating osteoblasts through the activator protein-1 and the runt domain binding sites

NASA Technical Reports Server (NTRS)

Winchester, S. K.; Selvamurugan, N.; D'Alonzo, R. C.; Partridge, N. C.

2000-01-01

Collagenase-3 mRNA is initially detectable when osteoblasts cease proliferation, increasing during differentiation and mineralization. We showed that this developmental expression is due to an increase in collagenase-3 gene transcription. Mutation of either the activator protein-1 or the runt domain binding site decreased collagenase-3 promoter activity, demonstrating that these sites are responsible for collagenase-3 gene transcription. The activator protein-1 and runt domain binding sites bind members of the activator protein-1 and core-binding factor family of transcription factors, respectively. We identified core-binding factor a1 binding to the runt domain binding site and JunD in addition to a Fos-related antigen binding to the activator protein-1 site. Overexpression of both c-Fos and c-Jun in osteoblasts or core-binding factor a1 increased collagenase-3 promoter activity. Furthermore, overexpression of c-Fos, c-Jun, and core-binding factor a1 synergistically increased collagenase-3 promoter activity. Mutation of either the activator protein-1 or the runt domain binding site resulted in the inability of c-Fos and c-Jun or core-binding factor a1 to increase collagenase-3 promoter activity, suggesting that there is cooperative interaction between the sites and the proteins. Overexpression of Fra-2 and JunD repressed core-binding factor a1-induced collagenase-3 promoter activity. Our results suggest that members of the activator protein-1 and core-binding factor families, binding to the activator protein-1 and runt domain binding sites are responsible for the developmental regulation of collagenase-3 gene expression in osteoblasts.

A Protocol for the Preparation of Cryoprecipitate and Cryo-depleted Plasma for Proteomic Studies.

PubMed

Sparrow, Rosemary L; Simpson, Richard J; Greening, David W

2017-01-01

Cryoprecipitate is a concentrate of high-molecular-weight plasma proteins that precipitate when frozen plasma is slowly thawed at 1-6 °C. The concentrate contains factor VIII (antihemophilic factor), von Willebrand factor (vWF), fibrinogen, factor XIII, fibronectin, and small amounts of other plasma proteins. Clinical grade preparations of cryoprecipitate are mainly used to treat fibrinogen deficiency caused by acute bleeding or functional abnormalities of the fibrinogen protein. In the past, cryoprecipitate was used to treat von Willebrand disease and hemophilia A (factor VIII deficiency), but the availability of more highly purified coagulation factor concentrates or recombinant protein preparations has superseded the use of cryoprecipitate for these coagulopathies. Cryo-depleted plasma ("cryosupernatant") is the plasma supernatant remaining following removal of the cryoprecipitate from frozen-thawed plasma. It contains all the other plasma proteins and clotting factors present in plasma that remain soluble during cold-temperature thawing of the plasma. This protocol describes the clinical-scale preparation of cryoprecipitate and cryo-depleted plasma for proteomic studies.

Endometrial proteins: a reappraisal.

PubMed

Seppälä, M; Julkunen, M; Riittinen, L; Koistinen, R

1992-06-01

Uterine factors influence reproduction at the macro-anatomy level, and the effects of hormonal steroids on endometrial morphology are well recognized in the histopathological diagnosis of dysfunctional bleeding and infertility. During the past decade, attention has been paid to endometrial protein synthesis and secretion with respect to endocrine stimuli and implantation, and to the paracrine/autocrine effects of endometrial peptide growth factors, their binding proteins and other factors. The emphasis of this presentation is on protein secretion of the secretory endometrium, in which progesterone plays a pivotal role. Insulin-like growth factors have receptors on the endometrium, and IGF-binding proteins, stimulated by progesterone, modulate the effects of IGFs locally. Also other protein products of the secretory endometrium have been reviewed in this communication, with special emphasis on studies of a progesterone-associated endometrial protein which has many names in the literature, such as PEP, PP14, alpha 2-PEG and AUP. Extensive studies are ongoing in many laboratories to elucidate the regulation, function, interplay at tissue and cellular levels, and clinical significance of these proteins.

Implementing a rational and consistent nomenclature for serine/arginine-rich protein splicing factors (SR proteins) in plants.

PubMed

Barta, Andrea; Kalyna, Maria; Reddy, Anireddy S N

2010-09-01

Growing interest in alternative splicing in plants and the extensive sequencing of new plant genomes necessitate more precise definition and classification of genes coding for splicing factors. SR proteins are a family of RNA binding proteins, which function as essential factors for constitutive and alternative splicing. We propose a unified nomenclature for plant SR proteins, taking into account the newly revised nomenclature of the mammalian SR proteins and a number of plant-specific properties of the plant proteins. We identify six subfamilies of SR proteins in Arabidopsis thaliana and rice (Oryza sativa), three of which are plant specific. The proposed subdivision of plant SR proteins into different subfamilies will allow grouping of paralogous proteins and simple assignment of newly discovered SR orthologs from other plant species and will promote functional comparisons in diverse plant species.

Structural and Functional Analysis of VQ Motif-Containing Proteins in Arabidopsis as Interacting Proteins of WRKY Transcription Factors1[W][OA

PubMed Central

Cheng, Yuan; Zhou, Yuan; Yang, Yan; Chi, Ying-Jun; Zhou, Jie; Chen, Jian-Ye; Wang, Fei; Fan, Baofang; Shi, Kai; Zhou, Yan-Hong; Yu, Jing-Quan; Chen, Zhixiang

2012-01-01

WRKY transcription factors are encoded by a large gene superfamily with a broad range of roles in plants. Recently, several groups have reported that proteins containing a short VQ (FxxxVQxLTG) motif interact with WRKY proteins. We have recently discovered that two VQ proteins from Arabidopsis (Arabidopsis thaliana), SIGMA FACTOR-INTERACTING PROTEIN1 and SIGMA FACTOR-INTERACTING PROTEIN2, act as coactivators of WRKY33 in plant defense by specifically recognizing the C-terminal WRKY domain and stimulating the DNA-binding activity of WRKY33. In this study, we have analyzed the entire family of 34 structurally divergent VQ proteins from Arabidopsis. Yeast (Saccharomyces cerevisiae) two-hybrid assays showed that Arabidopsis VQ proteins interacted specifically with the C-terminal WRKY domains of group I and the sole WRKY domains of group IIc WRKY proteins. Using site-directed mutagenesis, we identified structural features of these two closely related groups of WRKY domains that are critical for interaction with VQ proteins. Quantitative reverse transcription polymerase chain reaction revealed that expression of a majority of Arabidopsis VQ genes was responsive to pathogen infection and salicylic acid treatment. Functional analysis using both knockout mutants and overexpression lines revealed strong phenotypes in growth, development, and susceptibility to pathogen infection. Altered phenotypes were substantially enhanced through cooverexpression of genes encoding interacting VQ and WRKY proteins. These findings indicate that VQ proteins play an important role in plant growth, development, and response to environmental conditions, most likely by acting as cofactors of group I and IIc WRKY transcription factors. PMID:22535423

Structural and functional analysis of VQ motif-containing proteins in Arabidopsis as interacting proteins of WRKY transcription factors.

PubMed

Cheng, Yuan; Zhou, Yuan; Yang, Yan; Chi, Ying-Jun; Zhou, Jie; Chen, Jian-Ye; Wang, Fei; Fan, Baofang; Shi, Kai; Zhou, Yan-Hong; Yu, Jing-Quan; Chen, Zhixiang

2012-06-01

WRKY transcription factors are encoded by a large gene superfamily with a broad range of roles in plants. Recently, several groups have reported that proteins containing a short VQ (FxxxVQxLTG) motif interact with WRKY proteins. We have recently discovered that two VQ proteins from Arabidopsis (Arabidopsis thaliana), SIGMA FACTOR-INTERACTING PROTEIN1 and SIGMA FACTOR-INTERACTING PROTEIN2, act as coactivators of WRKY33 in plant defense by specifically recognizing the C-terminal WRKY domain and stimulating the DNA-binding activity of WRKY33. In this study, we have analyzed the entire family of 34 structurally divergent VQ proteins from Arabidopsis. Yeast (Saccharomyces cerevisiae) two-hybrid assays showed that Arabidopsis VQ proteins interacted specifically with the C-terminal WRKY domains of group I and the sole WRKY domains of group IIc WRKY proteins. Using site-directed mutagenesis, we identified structural features of these two closely related groups of WRKY domains that are critical for interaction with VQ proteins. Quantitative reverse transcription polymerase chain reaction revealed that expression of a majority of Arabidopsis VQ genes was responsive to pathogen infection and salicylic acid treatment. Functional analysis using both knockout mutants and overexpression lines revealed strong phenotypes in growth, development, and susceptibility to pathogen infection. Altered phenotypes were substantially enhanced through cooverexpression of genes encoding interacting VQ and WRKY proteins. These findings indicate that VQ proteins play an important role in plant growth, development, and response to environmental conditions, most likely by acting as cofactors of group I and IIc WRKY transcription factors.

Survey of rice proteins interacting with OsFCA and OsFY proteins which are homologous to the Arabidopsis flowering time proteins, FCA and FY.

PubMed

Jang, Yun Hee; Park, Hyo-Young; Kim, Soon-Kap; Lee, Jeong Hwan; Suh, Mi Chung; Chung, Young Soo; Paek, Kyung-Hee; Kim, Jeong-Kook

2009-08-01

The FCA protein is involved in controlling flowering time and plays more general roles in RNA-mediated chromatin silencing in Arabidopsis. It contains two RNA-binding domains and a WW domain. The FCA protein interacts with FY, a polyadenylation factor, via its WW domain. We previously characterized a rice gene, OsFCA, which was homologous to FCA. Here, we found that the OsFCA protein could interact through its WW domain with the following proteins: OsFY, a protein containing a CID domain present in RNA-processing factors such as Pcf11 and Nrd1; a protein similar to splicing factor SF1; a protein similar to FUSE splicing factor; and OsMADS8. The FY protein is associated with the 3' end processing machinery in Arabidopsis. Thus, we examined interactions between OsFY and the rice homologs (OsCstF-50, -64 and -77) of the AtCstF-50, -64 and -77 proteins. We found that OsFY could bind OsCstF50, whereas the OsCstF77 protein could bridge the interaction between OsCstF50 and OsCstF64. Taken together, our data suggest that OsFCA could interact with several proteins other than OsFY through its WW domain and may play several roles in rice.

Enhanced Functional Recombinant Factor VII Production by HEK 293 Cells Stably Transfected with VKORC1 where the Gamma-Carboxylase Inhibitor Calumenin is Stably Suppressed by shRNA Transfection

PubMed Central

Wajih, Nadeem; Owen, John; Wallin, Reidar

2008-01-01

Introduction Recombinant members of the vitamin K-dependent protein family (factors IX and VII and protein C) have become important pharmaceuticals in treatment of bleeding disorders and sepsis. However, because the in vivo γ-carboxylation system in stable cell lines used for transfection has a limited capacity of post translational γ carboxylation, the recovery of fully γ-carboxylated and functional proteins is low. Materials and Methods In this work we have engineered recombinant factor VII producing HEK 293 cells to stably overexpress VKORC1, the reduced vitamin K γ-carboxylase cofactor and in addition stably silenced the γ-carboxylase inhibitory protein calumenin. Results and Conclusions Stable cell lines transfected with only a factor VII cDNA had a 9% production of functional recombinant factor VII. On the other hand, these recombinant factor VII producing cells when engineered to overexpress VKORC1 and having calumenin stably suppressed more than 80% by shRNA expression, produced 68% functional factor VII. The technology presented should be applicable to all vertebrae members of the vitamin K-dependent protein family and should lower the production cost of the clinically used factors VII, IX and protein C. PMID:18177690

The effects of different preservation processes on the total protein and growth factor content in a new biological product developed from human amniotic membrane.

PubMed

Russo, Alessandra; Bonci, Paola; Bonci, Paolo

2012-06-01

The aim of this work is to quantify the total protein and growth factors content in a tissue-suspension obtained from processed human amniotic membrane (hAM). hAM was collected, frozen, freeze dried, powdered and sterilized by γ-irradiation. At each step of the process, samples were characterized for the total protein amounts by a Bradford protein assay and for the growth factor concentrations by ELISA test of the tissue suspensions. Frozen-hAM samples show higher release of total proteins and specific growth factors in the tissue suspension in comparison with freeze-dried hAM. We observed that even if the protein extraction is hindered once the tissue is dried, the powdering process allows a greater release in the tissue suspension of total proteins and growth factors after tissue re-solubilization in comparison with only the freeze-drying process (+91 ± 13% for EGF, +16 ± 4% for HGF, +11 ± 5% for FGF, +16 ± 9% for TGF-β1), and a greater release of EGF (85 ± 10%) in comparison with only the freezing process, because proteins become much readily solubilized in the solution. According with these results, we describe a protocol to obtain a new sterile biological product from hAM tissue, with well-known effects of thermal, mechanical and physical processes on the total protein and grow factors contents.

Direct interaction between the tobacco mosaic virus helicase domain and the ATP-bound resistance protein, N factor during the hypersensitive response in tobacco plants.

PubMed

Ueda, Hirokazu; Yamaguchi, Yube; Sano, Hiroshi

2006-05-01

Plants cope with pathogens with distinct mechanisms. One example is a gene-for-gene system, in which plants recognize the pathogen molecule by specified protein(s), this being called the R factor. However, mechanisms of interaction between proteins from the host and the pathogen are not completely understood. Here, we analyzed the mode of interaction between the N factor, a tobacco R factor, and the helicase domain (p50) of tobacco mosaic virus (TMV). To this end, domain dissected proteins were prepared and subjected to Agroinfiltration into intact leaves, followed by yeast two hybrid and pull-down assays. The results pointed to three novel features. First, the N factor was found to directly bind to the p50 of TMV, second, ATP was pre-requisite for this interaction, with formation of an ATP/N factor complex, and third, the N factor was shown to possess ATPase activity, which is enhanced by the p50. Moreover, we found that intra- and/or inter-molecular interactions take place in the N factor molecule. This interaction required ATP, and was disrupted by the p50. Based on these results, we propose a following model for the TMV recognition mechanism in tobacco plants. The N factor forms a complex with ATP, to which the helicase domain interacts, and enhances ATP hydrolysis. The resulting ADP/N factor complex then changes its conformation, thereby facilitating further interaction with the down-stream signaling factor(s). This model is consistent with the idea of 'protein machine'.

Proteopedia: 3D Visualization and Annotation of Transcription Factor-DNA Readout Modes

ERIC Educational Resources Information Center

Dantas Machado, Ana Carolina; Saleebyan, Skyler B.; Holmes, Bailey T.; Karelina, Maria; Tam, Julia; Kim, Sharon Y.; Kim, Keziah H.; Dror, Iris; Hodis, Eran; Martz, Eric; Compeau, Patricia A.; Rohs, Remo

2012-01-01

3D visualization assists in identifying diverse mechanisms of protein-DNA recognition that can be observed for transcription factors and other DNA binding proteins. We used Proteopedia to illustrate transcription factor-DNA readout modes with a focus on DNA shape, which can be a function of either nucleotide sequence (Hox proteins) or base pairing…

Role of growth hormone, insulin-like growth factor-I, and insulin-like growth factor binding proteins in the catabolic response to injury and infection.

PubMed

Lang, Charles H; Frost, Robert A

2002-05-01

The erosion of lean body mass resulting from protracted critical illness remains a significant risk factor for increased morbidity and mortality in this patient population. Previous studies have documented the well known impairment in nitrogen balance results from both an increase in muscle protein degradation as well as a decreased rate of both myofibrillar and sacroplasmic protein synthesis. This protein imbalance may be caused by an increased presence or activity of various catabolic agents, such as tumor necrosis factor-alpha, interleukin-1 beta, interleukin-6 or glucocorticoids, or may be mediated via a decreased concentration or responsiveness to various anabolic hormones, such as growth hormone or insulin-like growth factor-I. This review focuses on recent developments pertaining to the importance of alterations in the growth hormone-insulin-like growth factor-I axis as a mechanism for the observed defects in muscle protein balance.

WRKY transcription factors.

PubMed

Rushton, Paul J; Somssich, Imre E; Ringler, Patricia; Shen, Qingxi J

2010-05-01

WRKY transcription factors are one of the largest families of transcriptional regulators in plants and form integral parts of signalling webs that modulate many plant processes. Here, we review recent significant progress in WRKY transcription factor research. New findings illustrate that WRKY proteins often act as repressors as well as activators, and that members of the family play roles in both the repression and de-repression of important plant processes. Furthermore, it is becoming clear that a single WRKY transcription factor might be involved in regulating several seemingly disparate processes. Mechanisms of signalling and transcriptional regulation are being dissected, uncovering WRKY protein functions via interactions with a diverse array of protein partners, including MAP kinases, MAP kinase kinases, 14-3-3 proteins, calmodulin, histone deacetylases, resistance proteins and other WRKY transcription factors. WRKY genes exhibit extensive autoregulation and cross-regulation that facilitates transcriptional reprogramming in a dynamic web with built-in redundancy. 2010 Elsevier Ltd. All rights reserved.

Impact of antinutritional factors in food proteins on the digestibility of protein and the bioavailability of amino acids and on protein quality.

PubMed

Sarwar Gilani, G; Wu Xiao, Chao; Cockell, Kevin A

2012-08-01

Dietary antinutritional factors have been reported to adversely affect the digestibility of protein, bioavailability of amino acids and protein quality of foods. Published data on these negative effects of major dietary antinutritional factors are summarized in this manuscript. Digestibility and the quality of mixed diets in developing countries are considerably lower than of those in developed regions. For example, the digestibility of protein in traditional diets from developing countries such as India, Guatemala and Brazil is considerably lower compared to that of protein in typical North American diets (54-78 versus 88-94 %). Poor digestibility of protein in the diets of developing countries, which are based on less refined cereals and grain legumes as major sources of protein, is due to the presence of less digestible protein fractions, high levels of insoluble fibre, and/or high concentrations of antinutritional factors present endogenously or formed during processing. Examples of naturally occurring antinutritional factors include glucosinolates in mustard and canola protein products, trypsin inhibitors and haemagglutinins in legumes, tannins in legumes and cereals, gossypol in cottonseed protein products, and uricogenic nucleobases in yeast protein products. Heat/alkaline treatments of protein products may yield Maillard reaction compounds, oxidized forms of sulphur amino acids, D-amino acids and lysinoalanine (LAL, an unnatural nephrotoxic amino acid derivative). Among common food and feed protein products, soyabeans are the most concentrated source of trypsin inhibitors. The presence of high levels of dietary trypsin inhibitors from soyabeans, kidney beans or other grain legumes have been reported to cause substantial reductions in protein and amino acid digestibility (up to 50 %) and protein quality (up to 100 %) in rats and/or pigs. Similarly, the presence of high levels of tannins in sorghum and other cereals, fababean and other grain legumes can cause significant reductions (up to 23 %) in protein and amino acid digestibility in rats, poultry, and pigs. Normally encountered levels of phytates in cereals and legumes can reduce protein and amino acid digestibility by up to 10 %. D-amino acids and LAL formed during alkaline/heat treatment of lactalbumin, casein, soya protein or wheat protein are poorly digestible (less than 40 %), and their presence can reduce protein digestibility by up to 28 % in rats and pigs, and can cause a drastic reduction (100 %) in protein quality, as measured by rat growth methods. The adverse effects of antinutritional factors on protein digestibility and protein quality have been reported to be more pronounced in elderly rats (20-months old) compared to young (5-weeks old) rats, suggesting the use of old rats as a model for assessing the protein digestibility of products intended for the elderly.

The Effect of α-Mating Factor Secretion Signal Mutations on Recombinant Protein Expression in Pichia pastoris

PubMed Central

Lin-Cereghino, Geoff P.; Stark, Carolyn M.; Kim, Daniel; Chang, Jennifer; Shaheen, Nadia; Poerwanto, Hansel; Agari, Kimiko; Moua, Pachai; Low, Lauren K.; Tran, Namphuong; Huang, Amy D.; Nattestad, Maria; Oshiro, Kristin T.; Chang, John William; Chavan, Archana; Tsai, Jerry W.; Lin-Cereghino, Joan

2013-01-01

The methylotrophic yeast, Pichia pastoris, has been genetically engineered to produce many heterologous proteins for industrial and research purposes. In order to secrete proteins for easier purification from the extracellular medium, the coding sequence of recombinant proteins are initially fused to the Saccharomyces cerevisiae α-mating factor secretion signal leader. Extensive site-directed mutagenesis of the prepro region of the α-mating factor secretion signal sequence was performed in order to determine the effects of various deletions and substitutions on expression. Though some mutations clearly dampened protein expression, deletion of amino acids 57-70, corresponding to the predicted 3rd alpha helix of α-mating factor secretion signal, increased secretion of reporter proteins horseradish peroxidase and lipase at least 50% in small-scale cultures. These findings raise the possibility that the secretory efficiency of the leader can be further enhanced in the future. PMID:23454485

Mapping transcription factor interactome networks using HaloTag protein arrays.

PubMed

Yazaki, Junshi; Galli, Mary; Kim, Alice Y; Nito, Kazumasa; Aleman, Fernando; Chang, Katherine N; Carvunis, Anne-Ruxandra; Quan, Rosa; Nguyen, Hien; Song, Liang; Alvarez, José M; Huang, Shao-Shan Carol; Chen, Huaming; Ramachandran, Niroshan; Altmann, Stefan; Gutiérrez, Rodrigo A; Hill, David E; Schroeder, Julian I; Chory, Joanne; LaBaer, Joshua; Vidal, Marc; Braun, Pascal; Ecker, Joseph R

2016-07-19

Protein microarrays enable investigation of diverse biochemical properties for thousands of proteins in a single experiment, an unparalleled capacity. Using a high-density system called HaloTag nucleic acid programmable protein array (HaloTag-NAPPA), we created high-density protein arrays comprising 12,000 Arabidopsis ORFs. We used these arrays to query protein-protein interactions for a set of 38 transcription factors and transcriptional regulators (TFs) that function in diverse plant hormone regulatory pathways. The resulting transcription factor interactome network, TF-NAPPA, contains thousands of novel interactions. Validation in a benchmarked in vitro pull-down assay revealed that a random subset of TF-NAPPA validated at the same rate of 64% as a positive reference set of literature-curated interactions. Moreover, using a bimolecular fluorescence complementation (BiFC) assay, we confirmed in planta several interactions of biological interest and determined the interaction localizations for seven pairs. The application of HaloTag-NAPPA technology to plant hormone signaling pathways allowed the identification of many novel transcription factor-protein interactions and led to the development of a proteome-wide plant hormone TF interactome network.

Isolation and Identification of Proteins Secreted by Cells Cultured within Synthetic Hydrogel-Based Matrices

PubMed Central

2018-01-01

Cells interact with and remodel their microenvironment, degrading large extracellular matrix (ECM) proteins (e.g., fibronectin, collagens) and secreting new ECM proteins and small soluble factors (e.g., growth factors, cytokines). Synthetic mimics of the ECM have been developed as controlled cell culture platforms for use in both fundamental and applied studies. However, how cells broadly remodel these initially well-defined matrices remains poorly understood and difficult to probe. In this work, we have established methods for widely examining both large and small proteins that are secreted by cells within synthetic matrices. Specifically, human mesenchymal stem cells (hMSCs), a model primary cell type, were cultured within well-defined poly(ethylene glycol) (PEG)-peptide hydrogels, and these cell-matrix constructs were decellularized and degraded for subsequent isolation and analysis of deposited proteins. Shotgun proteomics using liquid chromatography and mass spectrometry identified a variety of proteins, including the large ECM proteins fibronectin and collagen VI. Immunostaining and confocal imaging confirmed these results and provided visualization of protein organization within the synthetic matrices. Additionally, culture medium was collected from the encapsulated hMSCs, and a Luminex assay was performed to identify secreted soluble factors, including vascular endothelial growth factor (VEGF), endothelial growth factor (EGF), basic fibroblast growth factor (FGF-2), interleukin 8 (IL-8), and tumor necrosis factor alpha (TNF-α). Together, these methods provide a unique approach for studying dynamic reciprocity between cells and synthetic microenvironments and have the potential to provide new biological insights into cell responses during three-dimensional (3D) controlled cell culture. PMID:29552635

Isolation and Identification of Proteins Secreted by Cells Cultured within Synthetic Hydrogel-Based Matrices.

PubMed

Sawicki, Lisa A; Choe, Leila H; Wiley, Katherine L; Lee, Kelvin H; Kloxin, April M

2018-03-12

Cells interact with and remodel their microenvironment, degrading large extracellular matrix (ECM) proteins (e.g., fibronectin, collagens) and secreting new ECM proteins and small soluble factors (e.g., growth factors, cytokines). Synthetic mimics of the ECM have been developed as controlled cell culture platforms for use in both fundamental and applied studies. However, how cells broadly remodel these initially well-defined matrices remains poorly understood and difficult to probe. In this work, we have established methods for widely examining both large and small proteins that are secreted by cells within synthetic matrices. Specifically, human mesenchymal stem cells (hMSCs), a model primary cell type, were cultured within well-defined poly(ethylene glycol) (PEG)-peptide hydrogels, and these cell-matrix constructs were decellularized and degraded for subsequent isolation and analysis of deposited proteins. Shotgun proteomics using liquid chromatography and mass spectrometry identified a variety of proteins, including the large ECM proteins fibronectin and collagen VI. Immunostaining and confocal imaging confirmed these results and provided visualization of protein organization within the synthetic matrices. Additionally, culture medium was collected from the encapsulated hMSCs, and a Luminex assay was performed to identify secreted soluble factors, including vascular endothelial growth factor (VEGF), endothelial growth factor (EGF), basic fibroblast growth factor (FGF-2), interleukin 8 (IL-8), and tumor necrosis factor alpha (TNF-α). Together, these methods provide a unique approach for studying dynamic reciprocity between cells and synthetic microenvironments and have the potential to provide new biological insights into cell responses during three-dimensional (3D) controlled cell culture.

Molecular switch-like regulation in motor proteins.

PubMed

Tafoya, Sara; Bustamante, Carlos

2018-06-19

Motor proteins are powered by nucleotide hydrolysis and exert mechanical work to carry out many fundamental biological tasks. To ensure their correct and efficient performance, the motors' activities are allosterically regulated by additional factors that enhance or suppress their NTPase activity. Here, we review two highly conserved mechanisms of ATP hydrolysis activation and repression operating in motor proteins-the glutamate switch and the arginine finger-and their associated regulatory factors. We examine the implications of these regulatory mechanisms in proteins that are formed by multiple ATPase subunits. We argue that the regulatory mechanisms employed by motor proteins display features similar to those described in small GTPases, which require external regulatory elements, such as dissociation inhibitors, exchange factors and activating proteins, to switch the protein's function 'on' and 'off'. Likewise, similar regulatory roles are taken on by the motor's substrate, additional binding factors, and even adjacent subunits in multimeric complexes. However, in motor proteins, more than one regulatory factor and the two mechanisms described here often underlie the machine's operation. Furthermore, ATPase regulation takes place throughout the motor's cycle, which enables a more complex function than the binary 'active' and 'inactive' states.This article is part of a discussion meeting issue 'Allostery and molecular machines'. © 2018 The Author(s).

Comparison of the structural basis for thermal stability between archaeal and bacterial proteins.

PubMed

Ding, Yanrui; Cai, Yujie; Han, Yonggang; Zhao, Bingqiang

2012-01-01

In this study, the structural basis for thermal stability in archaeal and bacterial proteins was investigated. There were many common factors that confer resistance to high temperature in both archaeal and bacterial proteins. These factors include increases in the Lys content, the bends and blanks of secondary structure, the Glu content of salt bridge; decreases in the number of main-side chain hydrogen bond and exposed surface area, and changes in the bends and blanks of amino acids. Certainly, the utilization of charged amino acids to form salt bridges is a primary factor. In both heat-resistant archaeal and bacterial proteins, most Glu and Asp participate in the formation of salt bridges. Other factors may influence either archaeal or bacterial protein thermostability, which includes the more frequent occurrence of shorter 3(10)-helices and increased hydrophobicity in heat-resistant archaeal proteins. However, there were increases in average helix length, the Glu content in salt bridges, temperature factors and decreases in the number of main-side chain hydrogen bonds, uncharged-uncharged hydrogen bonds, hydrophobicity, and buried and exposed polar surface area in heat-resistant bacterial proteins. Evidently, there are few similarities and many disparities between the heat-resistant mechanisms of archaeal and bacterial proteins.

Position Matters: Network Centrality Considerably Impacts Rates of Protein Evolution in the Human Protein–Protein Interaction Network

PubMed Central

Feyertag, Felix; Chakraborty, Sandip

2017-01-01

Abstract The proteins of any organism evolve at disparate rates. A long list of factors affecting rates of protein evolution have been identified. However, the relative importance of each factor in determining rates of protein evolution remains unresolved. The prevailing view is that evolutionary rates are dominantly determined by gene expression, and that other factors such as network centrality have only a marginal effect, if any. However, this view is largely based on analyses in yeasts, and accurately measuring the importance of the determinants of rates of protein evolution is complicated by the fact that the different factors are often correlated with each other, and by the relatively poor quality of available functional genomics data sets. Here, we use correlation, partial correlation and principal component regression analyses to measure the contributions of several factors to the variability of the rates of evolution of human proteins. For this purpose, we analyzed the entire human protein–protein interaction data set and the human signal transduction network—a network data set of exceptionally high quality, obtained by manual curation, which is expected to be virtually free from false positives. In contrast with the prevailing view, we observe that network centrality (measured as the number of physical and nonphysical interactions, betweenness, and closeness) has a considerable impact on rates of protein evolution. Surprisingly, the impact of centrality on rates of protein evolution seems to be comparable, or even superior according to some analyses, to that of gene expression. Our observations seem to be independent of potentially confounding factors and from the limitations (biases and errors) of interactomic data sets. PMID:28854629

The C Terminus of the Saccharomyces cerevisiae α-Factor Receptor Contributes to the Formation of Preactivation Complexes with Its Cognate G Protein

PubMed Central

Dosil, Mercedes; Schandel, Kimberly A.; Gupta, Ekta; Jenness, Duane D.; Konopka, James B.

2000-01-01

Binding of the α-factor pheromone to its G-protein-coupled receptor (encoded by STE2) activates the mating pathway in MATa yeast cells. To investigate whether specific interactions between the receptor and the G protein occur prior to ligand binding, we analyzed dominant-negative mutant receptors that compete with wild-type receptors for G proteins, and we analyzed the ability of receptors to suppress the constitutive signaling activity of mutant Gα subunits in an α-factor-independent manner. Although the amino acid substitution L236H in the third intracellular loop of the receptor impairs G-protein activation, this substitution had no influence on the ability of the dominant-negative receptors to sequester G proteins or on the ability of receptors to suppress the GPA1-A345T mutant Gα subunit. In contrast, removal of the cytoplasmic C-terminal domain of the receptor eliminated both of these activities even though the C-terminal domain is unnecessary for G-protein activation. Moreover, the α-factor-independent signaling activity of ste2-P258L mutant receptors was inhibited by the coexpression of wild-type receptors but not by coexpression of truncated receptors lacking the C-terminal domain. Deletion analysis suggested that the distal half of the C-terminal domain is critical for sequestration of G proteins. The C-terminal domain was also found to influence the affinity of the receptor for α-factor in cells lacking G proteins. These results suggest that the C-terminal cytoplasmic domain of the α-factor receptor, in addition to its role in receptor downregulation, promotes the formation of receptor–G-protein preactivation complexes. PMID:10866688

Aggregation factor analysis for protein formulation by a systematic approach using FTIR, SEC and design of experiments techniques.

PubMed

Feng, Yan Wen; Ooishi, Ayako; Honda, Shinya

2012-01-05

A simple systematic approach using Fourier transform infrared (FTIR) spectroscopy, size exclusion chromatography (SEC) and design of experiments (DOE) techniques was applied to the analysis of aggregation factors for protein formulations in stress and accelerated testings. FTIR and SEC were used to evaluate protein conformational and storage stabilities, respectively. DOE was used to determine the suitable formulation and to analyze both the main effect of single factors and the interaction effect of combined factors on aggregation. Our results indicated that (i) analysis at a low protein concentration is not always applicable to high concentration formulations; (ii) an investigation of interaction effects of combined factors as well as main effects of single factors is effective for improving conformational stability of proteins; (iii) with the exception of pH, the results of stress testing with regard to aggregation factors would be available for suitable formulation instead of performing time-consuming accelerated testing; (iv) a suitable pH condition should not be determined in stress testing but in accelerated testing, because of inconsistent effects of pH on conformational and storage stabilities. In summary, we propose a three-step strategy, using FTIR, SEC and DOE techniques, to effectively analyze the aggregation factors and perform a rapid screening for suitable conditions of protein formulation. Copyright © 2011 Elsevier B.V. All rights reserved.

Biochemical and proteomic analysis of spliceosome factors interacting with intron-1 of human papillomavirus type-16.

PubMed

Martínez-Salazar, Martha; López-Urrutia, Eduardo; Arechaga-Ocampo, Elena; Bonilla-Moreno, Raul; Martínez-Castillo, Macario; Díaz-Hernández, Job; Del Moral-Hernández, Oscar; Cedillo-Barrón, Leticia; Martines-Juarez, Víctor; De Nova-Ocampo, Monica; Valdes, Jesús; Berumen, Jaime; Villegas-Sepúlveda, Nicolás

2014-12-05

The human papillomavirus type 16 (HPV-16) E6/E7 spliced transcripts are heterogeneously expressed in cervical carcinoma. The heterogeneity of the E6/E7 splicing profile might be in part due to the intrinsic variation of splicing factors in tumor cells. However, the splicing factors that bind the E6/E7 intron 1 (In-1) have not been defined. Therefore, we aimed to identify these factors; we used HeLa nuclear extracts (NE) for in vitro spliceosome assembly. The proteins were allowed to bind to an RNA/DNA hybrid formed by the In-1 transcript and a 5'-biotinylated DNA oligonucleotide complementary to the upstream exon sequence, which prevented interference in protein binding to the intron. The hybrid probes bound with the nuclear proteins were coupled to streptavidin magnetic beads for chromatography affinity purification. Proteins were eluted and identified by mass spectrometry (MS). Approximately 170 proteins were identified by MS, 80% of which were RNA binding proteins, including canonical spliceosome core components, helicases and regulatory splicing factors. The canonical factors were identified as components of the spliceosomal B-complex. Although 35-40 of the identified factors were cognate splicing factors or helicases, they have not been previously detected in spliceosome complexes that were assembled using in vivo or in vitro models. Copyright © 2014 Elsevier B.V. All rights reserved.

How large B-factors can be in protein crystal structures.

PubMed

Carugo, Oliviero

2018-02-23

Protein crystal structures are potentially over-interpreted since they are routinely refined without any restraint on the upper limit of atomic B-factors. Consequently, some of their atoms, undetected in the electron density maps, are allowed to reach extremely large B-factors, even above 100 square Angstroms, and their final positions are purely speculative and not based on any experimental evidence. A strategy to define B-factors upper limits is described here, based on the analysis of protein crystal structures deposited in the Protein Data Bank prior 2008, when the tendency to allow B-factor to arbitrary inflate was limited. This B-factor upper limit (B_max) is determined by extrapolating the relationship between crystal structure average B-factor and percentage of crystal volume occupied by solvent (pcVol) to pcVol =100%, when, ab absurdo, the crystal contains only liquid solvent, the structure of which is, by definition, undetectable in electron density maps. It is thus possible to highlight structures with average B-factors larger than B_max, which should be considered with caution by the users of the information deposited in the Protein Data Bank, in order to avoid scientifically deleterious over-interpretations.

Polyphenol Compound as a Transcription Factor Inhibitor.

PubMed

Park, Seyeon

2015-10-30

A target-based approach has been used to develop novel drugs in many therapeutic fields. In the final stage of intracellular signaling, transcription factor-DNA interactions are central to most biological processes and therefore represent a large and important class of targets for human therapeutics. Thus, we focused on the idea that the disruption of protein dimers and cognate DNA complexes could impair the transcriptional activation and cell transformation regulated by these proteins. Historically, natural products have been regarded as providing the primary leading compounds capable of modulating protein-protein or protein-DNA interactions. Although their mechanism of action is not fully defined, polyphenols including flavonoids were found to act mostly as site-directed small molecule inhibitors on signaling. There are many reports in the literature of screening initiatives suggesting improved drugs that can modulate the transcription factor interactions responsible for disease. In this review, we focus on polyphenol compound inhibitors against dimeric forms of transcription factor components of intracellular signaling pathways (for instance, c-jun/c-fos (Activator Protein-1; AP-1), c-myc/max, Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and β-catenin/T cell factor (Tcf)).

A mechanism regulating proteolysis of specific proteins during renal tubular cell growth.

PubMed

Franch, H A; Sooparb, S; Du, J; Brown, N S

2001-06-01

Growth factors suppress the degradation of cellular proteins in lysosomes in renal epithelial cells. Whether this process also involves specific classes of proteins that influence growth processes is unknown. We investigated chaperone-mediated autophagy, a lysosomal import pathway that depends on the 73-kDa heat shock cognate protein and allows the degradation of proteins containing a specific lysosomal import consensus sequence (KFERQ motif). Epidermal growth factor (EGF) or ammonia, but not transforming growth factor beta1, suppresses total protein breakdown in cultured NRK-52E renal epithelial cells. EGF or ammonia prolonged the half-life of glyceraldehyde-3-phosphate dehydrogenase, a classic substrate for chaperone-mediated autophagy, by more than 90%, whereas transforming growth factor beta1 did not. EGF caused a similar increase in the half-life of the KFERQ-containing paired box-related transcription factor, Pax2. The increase in half-life was accompanied by an increased accumulation of proteins with a KFERQ motif including glyceraldehyde-3-phosphate dehydrogenase and Pax2. Ammonia also increased the level of the Pax2 protein. Lysosomal import of KFERQ proteins depends on the abundance of the 96-kDa lysosomal glycoprotein protein (lgp96), and we found that EGF caused a significant decrease in lgp96 in cellular homogenates and associated with lysosomes. We conclude that EGF in cultured renal cells regulates the breakdown of proteins targeted for destruction by chaperone-mediated autophagy. Because suppression of this pathway results in an increase in Pax2, these results suggest a novel mechanism for the regulation of cell growth.

[Experimental research on individual-specific rapid potassium supplementation strategy for fatal severe hypokalemia].

PubMed

Du, Yu; Mou, Yi; Liu, Jin

2018-05-01

To explore the effectiveness and safety of the individual-specific rapid potassium supplementation strategy, and to provide experimental basis for treating fatal severe hypokalemia. An acute fatal severe hypokalemia model was reproduced in 20 healthy adult Japanese big ear white rabbits with half lethal dose (LD50) of barium chloride (BaCl 2 ) solution 168 mg×5 mL -1 ×kg -1 . The rabbits were divided into conventional potassium supplementation group and individual-specific rapid potassium supplementation group according to random number table method with 10 rabbits in each group. All the animals were injected with 3% KCl through the auricular marginal veins by a micro-injection pump, and the target plasma potassium concentration was 4 mmol/L. The rabbits in conventional potassium supplementation group were administered continuously potassium infusion at the standard infusion rate of 0.4 mmol×kg -1 ×h -1 . And those in the individual-specific rapid potassium supplementation group were treated in two steps: first, a loading dose of potassium was rapidly injected within 5 minutes, and this step was repeated until the plasma potassium concentration increased to 3.5 mmol/L; second, a sustaining dose of potassium infusion was continued at the rate of 0.4 mmol×kg -1 ×h -1 after the increase in plasma potassium concentration. The changes in electrocardiogram, blood pressure, respiratory rate (RR), plasma potassium concentration, urine potassium concentration, urine volume, potassium content in extracellular fluid (ECF) and other parameters were monitored. The potassium supplementation, potassium excretion and potassium cross cell status were recorded. Adverse reactions and 7-day death were observed. Since the BaCl 2 administration, the plasma potassium concentration of all experimental rabbits were significantly lower than baseline at 0.5 hour, which was decreased below 2.5 mmol/L at 2.0 hours when the ventricular arrhythmias appeared, indicating the reproduction of fatal severe hypokalemia model was successful. There was no significant difference in gender, weight, baseline heart rate (HR), RR, mean arterial pressure (MAP), blood gas analysis or K + , Na + , Cl - levels between the two groups. Compared with baseline levels, MAP was significantly decreased and RR was significantly increased before potassium supplementation in both groups, but the parameters were improved significantly and restored to the baseline after potassium supplementation. There was no significant difference in MAP or RR during potassium supplementation between the two groups. The amount of potassium supplementation in two groups showed no significant differences. However, compared with the conventional potassium supplementation group, in the individual-specific rapid potassium supplementation group, the increase in plasma potassium concentration, urine potassium concentration, and the increase in potassium content in ECF were significantly increased [the increase in plasma potassium concentration (mmol/L): 2.40±0.33 vs. 1.51±0.75, urine potassium concentration (mmol/L): 164.94±18.07 vs. 108.35±19.67, the increase in potassium content in ECF (mmol): 1.17±0.16 vs. 0.73±0.35], the duration of potassium infusion was shortened (hours: 2.1±0.7 vs. 4.7±1.4), the total urine volume, renal excretion of potassium, and the amount of transcellular potassium shift were significantly decreased [total urine volume (mL): 6.40±1.78 vs. 13.60±4.69, renal excretion of potassium (mmol): 1.04±0.26 vs. 1.46±0.51, amount of transcellular potassium shift (mmol): 1.39±0.21 vs. 1.84±0.62], the duration of arrhythmia was shortened (minutes: 19.60±8.92 vs. 71.80±9.84), with statistically significant differences (all P < 0.05). Hyperkalemia did not occur in both groups. The rabbits of the individual-specific rapid potassium supplementation group were all alive, while 4 died in the conventional potassium supplementation group, and statistically significant difference was found between the two groups (P < 0.01). These data demonstrate that the individual-specific rapid potassium supplementation strategy can shorten the time for correcting hypokalemia, which is a better option to reverse life-threatening arrhythmia caused by severe hypokalemia, with a high rescue success rate. The process of potassium supplement is safe and effective.

Over-expression and purification strategies for recombinant multi-protein oligomers: a case study of Mycobacterium tuberculosis σ/anti-σ factor protein complexes.

PubMed

Thakur, Krishan Gopal; Jaiswal, Ravi Kumar; Shukla, Jinal K; Praveena, T; Gopal, B

2010-12-01

The function of a protein in a cell often involves coordinated interactions with one or several regulatory partners. It is thus imperative to characterize a protein both in isolation as well as in the context of its complex with an interacting partner. High resolution structural information determined by X-ray crystallography and Nuclear Magnetic Resonance offer the best route to characterize protein complexes. These techniques, however, require highly purified and homogenous protein samples at high concentration. This requirement often presents a major hurdle for structural studies. Here we present a strategy based on co-expression and co-purification to obtain recombinant multi-protein complexes in the quantity and concentration range that can enable hitherto intractable structural projects. The feasibility of this strategy was examined using the σ factor/anti-σ factor protein complexes from Mycobacterium tuberculosis. The approach was successful across a wide range of σ factors and their cognate interacting partners. It thus appears likely that the analysis of these complexes based on variations in expression constructs and procedures for the purification and characterization of these recombinant protein samples would be widely applicable for other multi-protein systems. Copyright © 2010 Elsevier Inc. All rights reserved.

Structure of the Get3 targeting factor in complex with its membrane protein cargo

DOE PAGES

Mateja, Agnieszka; Paduch, Marcin; Chang, Hsin-Yang; ...

2015-03-06

Tail-anchored (TA) proteins are a physiologically important class of membrane proteins targeted to the endoplasmic reticulum by the conserved guided-entry of TA proteins (GET) pathway. During transit, their hydrophobic transmembrane domains (TMDs) are chaperoned by the cytosolic targeting factor Get3, but the molecular nature of the functional Get3-TA protein targeting complex remains unknown. In this paper, we reconstituted the physiologic assembly pathway for a functional targeting complex and showed that it comprises a TA protein bound to a Get3 homodimer. Crystal structures of Get3 bound to different TA proteins showed an α-helical TMD occupying a hydrophobic groove that spans themore » Get3 homodimer. Finally, our data elucidate the mechanism of TA protein recognition and shielding by Get3 and suggest general principles of hydrophobic domain chaperoning by cellular targeting factors.« less

Acting on Actin: Rac and Rho Played by Yersinia.

PubMed

Aepfelbacher, Martin; Wolters, Manuel

2017-01-01

Pathogenic bacteria of the genus Yersinia include Y. pestis-the agent of plaque-and two enteropathogens, Y. enterocolitica, and Y. pseudotuberculosis. These pathogens have developed an array of virulence factors aimed at manipulating Rho GTP-binding proteins and the actin cytoskeleton in host cells to cross the intestinal barrier and suppress the immune system. Yersinia virulence factors include outer membrane proteins triggering cell invasion by binding to integrins, effector proteins injected into host cells to manipulate Rho protein functions and a Rho protein-activating exotoxin. Here, we present an overview of how Yersinia and host factors are integrated in a regulatory network that orchestrates the subversion of host defense.

Intrinsic factor

MedlinePlus

Intrinsic factor is a protein that helps your intestines absorb vitamin B12. It is made by cells in the ... Intrinsic factor is a protein that helps your body absorb vitamin B12. Vitamin B12 is needed for red blood ...

Genetics Home Reference: familial cylindromatosis

MedlinePlus

... instructions for making a protein that helps regulate nuclear factor-kappa-B. Nuclear factor-kappa-B is a group of related ... to certain signals. In regulating the action of nuclear factor-kappa-B, the CYLD protein allows cells ...

The sigma factors of Bacillus subtilis.

PubMed Central

Haldenwang, W G

1995-01-01

The specificity of DNA-dependent RNA polymerase for target promotes is largely due to the replaceable sigma subunit that it carries. Multiple sigma proteins, each conferring a unique promoter preference on RNA polymerase, are likely to be present in all bacteria; however, their abundance and diversity have been best characterized in Bacillus subtilis, the bacterium in which multiple sigma factors were first discovered. The 10 sigma factors thus far identified in B. subtilis directly contribute to the bacterium's ability to control gene expression. These proteins are not merely necessary for the expression of those operons whose promoters they recognize; in many instances, their appearance within the cell is sufficient to activate these operons. This review describes the discovery of each of the known B. subtilis sigma factors, their characteristics, the regulons they direct, and the complex restrictions placed on their synthesis and activities. These controls include the anticipated transcriptional regulation that modulates the expression of the sigma factor structural genes but, in the case of several of the B. subtilis sigma factors, go beyond this, adding novel posttranslational restraints on sigma factor activity. Two of the sigma factors (sigma E and sigma K) are, for example, synthesized as inactive precursor proteins. Their activities are kept in check by "pro-protein" sequences which are cleaved from the precursor molecules in response to intercellular cues. Other sigma factors (sigma B, sigma F, and sigma G) are inhibited by "anti-sigma factor" proteins that sequester them into complexes which block their ability to form RNA polymerase holoenzymes. The anti-sigma factors are, in turn, opposed by additional proteins which participate in the sigma factors' release. The devices used to control sigma factor activity in B, subtilis may prove to be as widespread as multiple sigma factors themselves, providing ways of coupling sigma factor activation to environmental or physiological signals that cannot be readily joined to other regulatory mechanisms. PMID:7708009

A Kinome-Wide Small Interfering RNA Screen Identifies Proviral and Antiviral Host Factors in Severe Acute Respiratory Syndrome Coronavirus Replication, Including Double-Stranded RNA-Activated Protein Kinase and Early Secretory Pathway Proteins

PubMed Central

de Wilde, Adriaan H.; Wannee, Kazimier F.; Scholte, Florine E. M.; Goeman, Jelle J.; ten Dijke, Peter; Snijder, Eric J.

2015-01-01

ABSTRACT To identify host factors relevant for severe acute respiratory syndrome-coronavirus (SARS-CoV) replication, we performed a small interfering RNA (siRNA) library screen targeting the human kinome. Protein kinases are key regulators of many cellular functions, and the systematic knockdown of their expression should provide a broad perspective on factors and pathways promoting or antagonizing coronavirus replication. In addition to 40 proteins that promote SARS-CoV replication, our study identified 90 factors exhibiting an antiviral effect. Pathway analysis grouped subsets of these factors in specific cellular processes, including the innate immune response and the metabolism of complex lipids, which appear to play a role in SARS-CoV infection. Several factors were selected for in-depth validation in follow-up experiments. In cells depleted for the β2 subunit of the coatomer protein complex (COPB2), the strongest proviral hit, we observed reduced SARS-CoV protein expression and a >2-log reduction in virus yield. Knockdown of the COPB2-related proteins COPB1 and Golgi-specific brefeldin A-resistant guanine nucleotide exchange factor 1 (GBF1) also suggested that COPI-coated vesicles and/or the early secretory pathway are important for SARS-CoV replication. Depletion of the antiviral double-stranded RNA-activated protein kinase (PKR) enhanced virus replication in the primary screen, and validation experiments confirmed increased SARS-CoV protein expression and virus production upon PKR depletion. In addition, cyclin-dependent kinase 6 (CDK6) was identified as a novel antiviral host factor in SARS-CoV replication. The inventory of pro- and antiviral host factors and pathways described here substantiates and expands our understanding of SARS-CoV replication and may contribute to the identification of novel targets for antiviral therapy. IMPORTANCE Replication of all viruses, including SARS-CoV, depends on and is influenced by cellular pathways. Although substantial progress has been made in dissecting the coronavirus replicative cycle, our understanding of the host factors that stimulate (proviral factors) or restrict (antiviral factors) infection remains far from complete. To study the role of host proteins in SARS-CoV infection, we set out to systematically identify kinase-regulated processes that influence virus replication. Protein kinases are key regulators in signal transduction, controlling a wide variety of cellular processes, and many of them are targets of approved drugs and other compounds. Our screen identified a variety of hits and will form the basis for more detailed follow-up studies that should contribute to a better understanding of SARS-CoV replication and coronavirus-host interactions in general. The identified factors could be interesting targets for the development of host-directed antiviral therapy to treat infections with SARS-CoV or other pathogenic coronaviruses. PMID:26041291

The canonical nuclear factor-κB pathway regulates cell survival in a developmental model of spinal cord motoneurons.

PubMed

Mincheva, Stefka; Garcera, Ana; Gou-Fabregas, Myriam; Encinas, Mario; Dolcet, Xavier; Soler, Rosa M

2011-04-27

In vivo and in vitro motoneuron survival depends on the support of neurotrophic factors. These factors activate signaling pathways related to cell survival or inactivate proteins involved in neuronal death. In the present work, we analyzed the involvement of the nuclear factor-κB (NF-κB) pathway in mediating mouse spinal cord motoneuron survival promoted by neurotrophic factors. This pathway comprises ubiquitously expressed transcription factors that could be activated by two different routes: the canonical pathway, associated with IKKα/IKKβ kinase phosphorylation and nuclear translocation RelA (p65)/p50 transcription factors; and the noncanonical pathway, related to IKKα kinase homodimer phosphorylation and RelB/p52 transcription factor activation. In our system, we show that neurotrophic factors treatment induced IKKα and IKKβ phosphorylation and RelA nuclear translocation, suggesting NF-κB pathway activation. Protein levels of different members of the canonical or noncanonical pathways were reduced in a primary culture of isolated embryonic motoneurons using an interference RNA approach. Even in the presence of neurotrophic factors, selective reduction of IKKα, IKKβ, or RelA proteins induced cell death. In contrast, RelB protein reduction did not have a negative effect on motoneuron survival. Together these results demonstrated that the canonical NF-κB pathway mediates motoneuron survival induced by neurotrophic factors, and the noncanonical pathway is not related to this survival effect. Canonical NF-κB blockade induced an increase of Bim protein level and apoptotic cell death. Bcl-x(L) overexpression or Bax reduction counteracted this apoptotic effect. Finally, RelA knockdown causes changes of CREB and Smn protein levels.

A Single Rainbow Trout Cobalamin-binding Protein Stands in for Three Human Binders

PubMed Central

Greibe, Eva; Fedosov, Sergey; Sorensen, Boe S.; Højrup, Peter; Poulsen, Steen S.; Nexo, Ebba

2012-01-01

Cobalamin uptake and transport in mammals are mediated by three cobalamin-binding proteins: haptocorrin, intrinsic factor, and transcobalamin. The nature of cobalamin-binding proteins in lower vertebrates remains to be elucidated. The aim of this study was to characterize the cobalamin-binding proteins of the rainbow trout (Oncorhynchus mykiss) and to compare their properties with those of the three human cobalamin-binding proteins. High cobalamin-binding capacity was found in trout stomach (210 pmol/g), roe (400 pmol/g), roe fluid (390 nmol/liter), and plasma (2500 nmol/liter). In all cases, it appeared to be the same protein based on analysis of partial sequences and immunological responses. The trout cobalamin-binding protein was purified from roe fluid, sequenced, and further characterized. Like haptocorrin, the trout cobalamin-binding protein was stable at low pH and had a high binding affinity for the cobalamin analog cobinamide. Like haptocorrin and transcobalamin, the trout cobalamin-binding protein was present in plasma and recognized ligands with altered nucleotide moiety. Like intrinsic factors, the trout cobalamin-binding protein was present in the stomach and resisted degradation by trypsin and chymotrypsin. It also resembled intrinsic factor in the composition of conserved residues in the primary cobalamin-binding site in the C terminus. The trout cobalamin-binding protein was glycosylated and displayed spectral properties comparable with those of haptocorrin and intrinsic factor. In conclusion, only one soluble cobalamin-binding protein was identified in the rainbow trout, a protein that structurally behaves like an intermediate between the three human cobalamin-binding proteins. PMID:22872637

BAR domain proteins regulate Rho GTPase signaling.

PubMed

Aspenström, Pontus

2014-01-01

BAR proteins comprise a heterogeneous group of multi-domain proteins with diverse biological functions. The common denominator is the Bin-Amphiphysin-Rvs (BAR) domain that not only confers targeting to lipid bilayers, but also provides scaffolding to mold lipid membranes into concave or convex surfaces. This function of BAR proteins is an important determinant in the dynamic reconstruction of membrane vesicles, as well as of the plasma membrane. Several BAR proteins function as linkers between cytoskeletal regulation and membrane dynamics. These links are provided by direct interactions between BAR proteins and actin-nucleation-promoting factors of the Wiskott-Aldrich syndrome protein family and the Diaphanous-related formins. The Rho GTPases are key factors for orchestration of this intricate interplay. This review describes how BAR proteins regulate the activity of Rho GTPases, as well as how Rho GTPases regulate the function of BAR proteins. This mutual collaboration is a central factor in the regulation of vital cellular processes, such as cell migration, cytokinesis, intracellular transport, endocytosis, and exocytosis.

The mammalian iris-ciliary complex affects organization and synthesis of cytoskeletal proteins of organ and tissue cultured lens epithelial cells.

PubMed

Banerjee, A; Emanuel, K; Parafina, J; Bagchi, M

1992-10-01

A water soluble growth inhibitor was isolated from the mammalian ocular iris-ciliary complex. The molecular weight of this protein is 10 kD or lower as determined by ultrafiltration fractionation. The iris-ciliary (IC) complex water soluble protein(s) significantly inhibits synthesis of lower molecular weight proteins of the epithelial cells of the organ cultured mammalian ocular lens. It was also found that this inhibitory effect of IC is mediated via the structural organization of the lens. Monolayer cultures of the lens epithelial cells exposed to IC did not manifest any inhibition of their protein synthesis. Moreover, these tissue cultured lens epithelial (TCLE) cells showed a significant increase in their protein synthetic activities in response to the presence of IC factors in the culture medium. It is postulated that the IC activity is modulated via either the lens capsule, an extracellular matrix, or due to the specific organization of the intact lens. The specific effects of IC on the cytoskeletal organization and synthesis in the organ cultured lens epithelial (OCLE) and TCLE cells were also examined. Both groups, treated with IC factors, manifested significant alterations in their protein synthetic activities and cytoskeletal architecture. The 3H-leucine incorporation experiments showed that alpha-actin and alpha-tubulin synthesis is partially inhibited by IC factors in OCLE cells but vimentin synthesis is not, whereas in TCLE cells all of them showed increased synthesis in response to IC factors. Turnover rates of these proteins in both OCLE and TCLE cells were also computed. The immunofluorescence and microscopic evaluation of OCLE and TCLE cells exposed to IC factors illustrated significant alteration in the cytoarchitecture of the filaments. We demonstrate that an inhibitor(s) molecule of 10 kD or lower size isolated from IC inhibited protein synthesis of OCLE cells and stimulated protein synthesis in TCLE cells. The IC factor also affects the synthesis and organization of cytoskeletal filaments of both the OCLE and TCLE cells.

First Light for ASTROVIRTEL Project

NASA Astrophysics Data System (ADS)

2000-04-01

Astronomical data archives increasingly resemble virtual gold mines of information. A new project, known as ASTROVIRTEL aims to exploit these astronomical treasure troves by allowing scientists to use the archives as virtual telescopes. The competition for observing time on large space- and ground-based observatories such as the ESA/NASA Hubble Space Telescope and the ESO Very Large Telescope (VLT) is intense. On average, less than a quarter of applications for observing time are successful. The fortunate scientist who obtains observing time usually has one year of so-called proprietary time to work with the data before they are made publicly accessible and can be used by other astronomers. Precious data from these large research facilities retain their value far beyond their first birthday and may still be useful decades after they were first collected. The enormous quantity of valuable astronomical data now stored in the archives of the European Southern Observatory (ESO) and the Space Telescope-European Coordinating Facility (ST-ECF) is increasingly attracting the attention of astronomers. Scientists are aware that one set of observations can serve many different scientific purposes, including some that were not considered at all when the observations were first made. Data archives as "gold mines" for research [ASTROVIRTEL Logo; JPEG - 184 k] Astronomical data archives increasingly resemble virtual gold mines of information. A new project, known as ASTROVIRTEL or "Accessing Astronomical Archives as Virtual Telescopes" aims to exploit these astronomical treasure troves. It is supported by the European Commission (EC) within the "Access to Research Infrastructures" action under the "Improving Human Potential & the Socio-economic Knowledge Base" of the EC (under EU Fifth Framework Programme). ASTROVIRTEL has been established on behalf of the European Space Agency (ESA) and the European Southern Observatory (ESO) in response to rapid developments currently taking place in the fields of telescope and detector construction, computer hardware, data processing, archiving, and telescope operation. Nowadays astronomical telescopes can image increasingly large areas of the sky. They use more and more different instruments and are equipped with ever-larger detectors. The quantity of astronomical data collected is rising dramatically, generating a corresponding increase in potentially interesting research projects. These large collections of valuable data have led to the useful concept of "data mining", whereby large astronomical databases are exploited to support original research. However, it has become obvious that scientists need additional support to cope efficiently with the massive amounts of data available and so to exploit the true potential of the databases. The strengths of ASTROVIRTEL ASTROVIRTEL is the first virtual astronomical telescope dedicated to data mining. It is currently being established at the joint ESO/Space Telescope-European Coordinating Facility Archive in Garching (Germany). Scientists from EC member countries and associated states will be able to apply for support for a scientific project based on access to and analysis of data from the Hubble Space Telescope (HST), Very Large Telescope (VLT), New Technology Telescope (NTT), and Wide Field Imager (WFI) archives, as well as a number of other related archives, including the Infrared Space Observatory (ISO) archive. Scientists will be able to visit the archive site and collaborate with the archive specialists there. Special software tools that incorporate advanced methods for exploring the enormous quantities of information available will be developed. Statements The project co-ordinator, Piero Benvenuti , Head of ST-ECF, elaborates on the advantages of ASTROVIRTEL: "The observations by the ESA/NASA Hubble Space Telescope and, more recently, by the ESO Very Large Telescope, have already been made available on-line to the astronomical community, once the proprietary period of one year has elapsed. ASTROVIRTEL is different, in that astronomers are now invited to regard the archive as an "observatory" in its own right: a facility that, when properly used, may provide an answer to their specific scientific questions. The architecture of the archives as well as their suite of software tools may have to evolve to respond to the new demand. ASTROVIRTEL will try to drive this evolution on the basis of the scientific needs of its users." Peter Quinn , the Head of ESO's Data Management and Operations Division, is of the same opinion: "The ESO/HST Archive Facility at ESO Headquarters in Garching is currently the most rapidly growing astronomical archive resource in the world. This archive is projected to contain more than 100 Terabytes (100,000,000,000,000 bytes) of data within the next four years. The software and hardware technologies for the archive will be jointly developed and operated by ESA and ESO staff and will be common to both HST and ESO data archives. The ASTROVIRTEL project will provide us with real examples of scientific research programs that will push the capabilities of the archive and allow us to identify and develop new software tools for data mining. The growing archive facility will provide the European astronomical community with new digital windows on the Universe." Note [1] This is a joint Press Release by the European Southern Observatory (ESO) and the Space Telescope European Coordinating Facility (ST-ECF). Additional information More information about ASTROVIRTEL can be found at the dedicated website at: http://www.stecf.org/astrovirtel The European Southern Observatory (ESO) is an intergovernmental organisation, supported by eight European countries: Belgium, Denmark, France, Germany, Italy, The Netherlands, Sweden and Switzerland. The European Space Agency is an intergovernmental organisation supported by 15 European countries: Austria, Belgium, Denmark, Finland, France, Germany, Ireland, Italy, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and the United Kingdom. The Space Telescope European Coordinating Facility (ST-ECF) is a co-operation between the European Space Agency and the European Southern Observatory. The Hubble Space Telescope (HST) is a project of international co-operation between NASA and ESA.

Protein Z efficiently depletes thrombin generation in disseminated intravascular coagulation with poor prognosis.

PubMed

Lee, Nuri; Kim, Ji-Eun; Gu, Ja-Yoon; Yoo, Hyun Ju; Kim, Inho; Yoon, Sung-Soo; Park, Seonyang; Han, Kyou-Sup; Kim, Hyun Kyung

2016-01-01

Disseminated intravascular coagulation (DIC) is characterized by consumption of coagulation factors and anticoagulants. Thrombin generation assay (TGA) gives useful information about global hemostatic status. We developed a new TGA system that anticoagulant addition can deplete thrombin generation in plasma, which may reflect defective anticoagulant system in DIC. TGAs were measured on the calibrated automated thrombogram with and without thrombomodulin or protein Z in 152 patients who were suspected of having DIC, yielding four parameters including lag time, endogenous thrombin potential, peak thrombin and time-to-peak in each experiment. Nonsurvivors showed significantly prolonged lag time and time-to-peak in TGA-protein Z system, which was performed with added protein Z. In multivariate Cox regression analysis, lag time and time-to-peak in TGA system were significant independent prognostic factors. In TGA-protein Z system, lag time and time-to-peak were revealed as independent prognostic factors of DIC. Protein Z addition could potentiate its anticoagulant effect in DIC with poor prognosis, suggesting the presence of defective protein Z system. The prolonged lag time and time-to-peak in both TGA and TGA-protein Z systems are expected to be used as independent prognostic factors of DIC.

Engineering a pharmacologically superior form of granulocyte-colony-stimulating factor by fusion with gelatin-like-protein polymer.

PubMed

Huang, Yan-Shan; Wen, Xiao-Fang; Wu, Yi-Liang; Wang, Ye-Fei; Fan, Min; Yang, Zhi-Yu; Liu, Wei; Zhou, Lin-Fu

2010-03-01

The plasma half-life of therapeutic proteins is a critical factor in many clinical applications. Therefore, new strategies to prolong plasma half-life of long-acting peptides and protein drugs are in high demand. Here, we designed an artificial gelatin-like protein (GLK) and fused this hydrophilic GLK polymer to granulocyte-colony-stimulating factor (G-CSF) to generate a chimeric GLK/G-CSF fusion protein. The genetically engineered recombinant GLK/G-CSF (rGLK/G-CSF) fusion protein was purified from Pichia pastoris. In vitro studies demonstrated that rGLK/G-CSF possessed an enlarged hydrodynamic radius, improved thermal stability and retained full bioactivity compared to unfused G-CSF. Following a single subcutaneous administration to rats, the rGLK/G-CSF fusion protein displayed a slower plasma clearance rate and stimulated greater and longer lasting increases in circulating white blood cells than G-CSF. Our findings indicate that fusion with this artificial, hydrophilic, GLK polymer provides many advantages in the construction of a potent hematopoietic factor with extended plasma half-life. This approach could be easily applied to other therapeutic proteins and have important clinical applications. (c) 2009 Elsevier B.V. All rights reserved.

Use of multiple picosecond high-mass molecular dynamics simulations to predict crystallographic B-factors of folded globular proteins.

PubMed

Pang, Yuan-Ping

2016-09-01

Predicting crystallographic B-factors of a protein from a conventional molecular dynamics simulation is challenging, in part because the B-factors calculated through sampling the atomic positional fluctuations in a picosecond molecular dynamics simulation are unreliable, and the sampling of a longer simulation yields overly large root mean square deviations between calculated and experimental B-factors. This article reports improved B-factor prediction achieved by sampling the atomic positional fluctuations in multiple picosecond molecular dynamics simulations that use uniformly increased atomic masses by 100-fold to increase time resolution. Using the third immunoglobulin-binding domain of protein G, bovine pancreatic trypsin inhibitor, ubiquitin, and lysozyme as model systems, the B-factor root mean square deviations (mean ± standard error) of these proteins were 3.1 ± 0.2-9 ± 1 Å 2 for Cα and 7.3 ± 0.9-9.6 ± 0.2 Å 2 for Cγ, when the sampling was done for each of these proteins over 20 distinct, independent, and 50-picosecond high-mass molecular dynamics simulations with AMBER forcefield FF12MC or FF14SB. These results suggest that sampling the atomic positional fluctuations in multiple picosecond high-mass molecular dynamics simulations may be conducive to a priori prediction of crystallographic B-factors of a folded globular protein.

Protein stability: a crystallographer’s perspective

DOE Office of Scientific and Technical Information (OSTI.GOV)

Deller, Marc C., E-mail: mdeller@stanford.edu; Kong, Leopold; Rupp, Bernhard

An understanding of protein stability is essential for optimizing the expression, purification and crystallization of proteins. In this review, discussion will focus on factors affecting protein stability on a somewhat practical level, particularly from the view of a protein crystallographer. Protein stability is a topic of major interest for the biotechnology, pharmaceutical and food industries, in addition to being a daily consideration for academic researchers studying proteins. An understanding of protein stability is essential for optimizing the expression, purification, formulation, storage and structural studies of proteins. In this review, discussion will focus on factors affecting protein stability, on a somewhatmore » practical level, particularly from the view of a protein crystallographer. The differences between protein conformational stability and protein compositional stability will be discussed, along with a brief introduction to key methods useful for analyzing protein stability. Finally, tactics for addressing protein-stability issues during protein expression, purification and crystallization will be discussed.« less

A systematic analysis of factors localized to damaged chromatin reveals PARP-dependent recruitment of transcription factors

PubMed Central

Izhar, Lior; Adamson, Britt; Ciccia, Alberto; Lewis, Jedd; Pontano-Vaites, Laura; Leng, Yumei; Liang, Anthony C.; Westbrook, Thomas F.; Harper, J. Wade; Elledge, Stephen J.

2015-01-01

Localization to sites of DNA damage is a hallmark of DNA damage response (DDR) proteins. To identify new DDR factors, we screened epitope-tagged proteins for localization to sites of chromatin damaged by UV laser microirradiation and found >120 proteins that localize to damaged chromatin. These include the BAF tumor suppressor complex and the ALS candidate protein TAF15. TAF15 contains multiple domains that bind damaged chromatin in a PARP-dependent manner, suggesting a possible role as glue that tethers multiple PAR chains together. Many positives were transcription factors and >70% of randomly tested transcription factors localized to sites of DNA damage and approximately 90% were PARP-dependent for localization. Mutational analyses showed that localization to damaged chromatin is DNA-binding domain-dependent. By examining Hoechst staining patterns at damage sites, we see evidence of chromatin decompaction that is PARP-dependent. We propose that PARP-regulated chromatin remodeling at sites of damage allows transient accessibility of DNA-binding proteins. PMID:26004182

Pharmacological targeting of the transcription factor SOX18 delays breast cancer in mice.

PubMed

Overman, Jeroen; Fontaine, Frank; Moustaqil, Mehdi; Mittal, Deepak; Sierecki, Emma; Sacilotto, Natalia; Zuegg, Johannes; Robertson, Avril Ab; Holmes, Kelly; Salim, Angela A; Mamidyala, Sreeman; Butler, Mark S; Robinson, Ashley S; Lesieur, Emmanuelle; Johnston, Wayne; Alexandrov, Kirill; Black, Brian L; Hogan, Benjamin M; De Val, Sarah; Capon, Robert J; Carroll, Jason S; Bailey, Timothy L; Koopman, Peter; Jauch, Ralf; Smyth, Mark J; Cooper, Matthew A; Gambin, Yann; Francois, Mathias

2017-01-31

Pharmacological targeting of transcription factors holds great promise for the development of new therapeutics, but strategies based on blockade of DNA binding, nuclear shuttling, or individual protein partner recruitment have yielded limited success to date. Transcription factors typically engage in complex interaction networks, likely masking the effects of specifically inhibiting single protein-protein interactions. Here, we used a combination of genomic, proteomic and biophysical methods to discover a suite of protein-protein interactions involving the SOX18 transcription factor, a known regulator of vascular development and disease. We describe a small-molecule that is able to disrupt a discrete subset of SOX18-dependent interactions. This compound selectively suppressed SOX18 transcriptional outputs in vitro and interfered with vascular development in zebrafish larvae. In a mouse pre-clinical model of breast cancer, treatment with this inhibitor significantly improved survival by reducing tumour vascular density and metastatic spread. Our studies validate an interactome-based molecular strategy to interfere with transcription factor activity, for the development of novel disease therapeutics.

Quantification of the environmental impact of different dietary protein choices.

PubMed

Reijnders, Lucas; Soret, Sam

2003-09-01

Quantitative environmental evaluations of meat, fresh vegetables, and processed protein based on soybeans suggest that the environmental burden of vegetarian foods is usually relatively low when production and processing are considered. The environmental comparison of cheese varieties made from cow milk and directly from lupine and the evaluation of energy inputs in fish protein and vegetable protein also suggest an environmental advantage for vegetarian food. In the evaluation of processed protein food based on soybeans and meat protein, a variety of environmental impacts associated with primary production and processing are a factor 4.4-> 100 to the disadvantage of meat. The comparison of cheese varieties gives differences in specific environmental impacts ranging between a factor 5 and 21. And energy use for fish protein may be up to a factor 14 more than for protein of vegetable origin. Assessment suggests that on average the complete life cycle environmental impact of nonvegetarian meals may be roughly a factor 1.5-2 higher than the effect of vegetarian meals in which meat has been replaced by vegetable protein. Although on average vegetarian diets may well have an environmental advantage, exceptions may also occur. Long-distance air transport, deep-freezing, and some horticultural practices may lead to environmental burdens for vegetarian foods exceeding those for locally produced organic meat.

Novel host restriction factors implicated in HIV-1 replication.

PubMed

Ghimire, Dibya; Rai, Madhu; Gaur, Ritu

2018-04-01

Human immunodeficiency virus-1 (HIV-1) is known to interact with multiple host cellular proteins during its replication in the target cell. While many of these host cellular proteins facilitate viral replication, a number of them are reported to inhibit HIV-1 replication at various stages of its life cycle. These host cellular proteins, which are known as restriction factors, constitute an integral part of the host's first line of defence against the viral pathogen. Since the discovery of apolipoprotein B mRNA-editing enzyme 3G (APOBEC3G) as an HIV-1 restriction factor, several human proteins have been identified that exhibit anti-HIV-1 restriction. While each restriction factor employs a distinct mechanism of inhibition, the HIV-1 virus has equally evolved complex counter strategies to neutralize their inhibitory effect. APOBEC3G, tetherin, sterile alpha motif and histidine-aspartate domain 1 (SAMHD1), and trim-5α are some of the best known HIV-1 restriction factors that have been studied in great detail. Recently, six novel restriction factors were discovered that exhibit significant antiviral activity: endoplasmic reticulum α1,2-mannosidase I (ERManI), translocator protein (TSPO), guanylate-binding protein 5 (GBP5), serine incorporator (SERINC3/5) and zinc-finger antiviral protein (ZAP). The focus of this review is to discuss the antiviral mechanism of action of these six restriction factors and provide insights into the probable counter-evasion strategies employed by the HIV-1 virus. The recent discovery of new restriction factors substantiates the complex host-pathogen interactions occurring during HIV-1 pathogenesis and makes it imperative that further investigations are conducted to elucidate the molecular basis of HIV-1 replication.

TALE transcription factors during early development of the vertebrate brain and eye.

PubMed

Schulte, Dorothea; Frank, Dale

2014-01-01

Our brain's cognitive performance arises from the coordinated activities of billions of nerve cells. Despite a high degree of morphological and functional differences, all neurons of the vertebrate central nervous system (CNS) arise from a common field of multipotent progenitors. Cell fate specification and differentiation are directed by multistep processes that include inductive/external cues, such as the extracellular matrix or growth factors, and cell-intrinsic determinants, such as transcription factors and epigenetic modulators of proteins and DNA. Here we review recent findings implicating TALE-homeodomain proteins in these processes. Although originally identified as HOX-cofactors, TALE proteins also contribute to many physiological processes that do not require HOX-activity. Particular focus is, therefore, given to HOX-dependent and -independent functions of TALE proteins during early vertebrate brain development. Additionally, we provide an overview about known upstream and downstream factors of TALE proteins in the developing vertebrate brain and discuss general concepts of how TALE proteins function to modulate neuronal cell fate specification. Copyright © 2013 Wiley Periodicals, Inc.

Methylation of eukaryotic elongation factor 2 induced by basic fibroblast growth factor via mitogen-activated protein kinase.

PubMed

Jung, Gyung Ah; Shin, Bong Shik; Jang, Yeon Sue; Sohn, Jae Bum; Woo, Seon Rang; Kim, Jung Eun; Choi, Go; Lee, Kyung Mi; Min, Bon Hong; Lee, Kee Ho; Park, Gil Hong

2011-10-31

Protein arginine methylation is important for a variety of cellular processes including transcriptional regulation, mRNA splicing, DNA repair, nuclear/cytoplasmic shuttling and various signal transduction pathways. However, the role of arginine methylation in protein biosynthesis and the extracellular signals that control arginine methylation are not fully understood. Basic fibroblast growth factor (bFGF) has been identified as a potent stimulator of myofibroblast dedifferentiation into fibroblasts. We demonstrated that symmetric arginine dimethylation of eukaryotic elongation factor 2 (eEF2) is induced by bFGF without the change in the expression level of eEF2 in mouse embryo fibroblast NIH3T3 cells. The eEF2 methylation is preceded by ras-raf-mitogen-activated protein kinase kinase (MEK)-extracellular signal-regulated kinase (ERK1/2)- p21Cip/WAF1 activation, and suppressed by the mitogenactivated protein kinase (MAPK) inhibitor PD98059 and p21Cip/WAF1 short interfering RNA (siRNA). We determined that protein arginine methyltransferase 7 (PRMT7) is responsible for the methylation, and that PRMT5 acts as a coordinator. Collectively, we demonstrated that eEF2, a key factor involved in protein translational elongation is symmetrically arginine-methylated in a reversible manner, being regulated by bFGF through MAPK signaling pathway.

Methylation of eukaryotic elongation factor 2 induced by basic fibroblast growth factor via mitogen-activated protein kinase

PubMed Central

Jung, Gyung Ah; Shin, Bong Shik; Jang, Yeon Sue; Sohn, Jae Bum; Woo, Seon Rang; Kim, Jung Eun; Choi, Go; Lee, Kyung-Mi; Min, Bon Hong

2011-01-01

Protein arginine methylation is important for a variety of cellular processes including transcriptional regulation, mRNA splicing, DNA repair, nuclear/cytoplasmic shuttling and various signal transduction pathways. However, the role of arginine methylation in protein biosynthesis and the extracellular signals that control arginine methylation are not fully understood. Basic fibroblast growth factor (bFGF) has been identified as a potent stimulator of myofibroblast dedifferentiation into fibroblasts. We demonstrated that symmetric arginine dimethylation of eukaryotic elongation factor 2 (eEF2) is induced by bFGF without the change in the expression level of eEF2 in mouse embryo fibroblast NIH3T3 cells. The eEF2 methylation is preceded by ras-raf-mitogen-activated protein kinase kinase (MEK)-extracellular signal-regulated kinase (ERK1/2)-p21Cip/WAF1 activation, and suppressed by the mitogen-activated protein kinase (MAPK) inhibitor PD98059 and p21Cip/WAF1 short interfering RNA (siRNA). We determined that protein arginine methyltransferase 7 (PRMT7) is responsible for the methylation, and that PRMT5 acts as a coordinator. Collectively, we demonstrated that eEF2, a key factor involved in protein translational elongation is symmetrically arginine-methylated in a reversible manner, being regulated by bFGF through MAPK signaling pathway. PMID:21778808

Production of functional human insulin-like growth factor binding proteins (IGFBPs) using recombinant expression in HEK293 cells.

PubMed

Wanscher, Anne Sofie Molsted; Williamson, Michael; Ebersole, Tasja Wainani; Streicher, Werner; Wikström, Mats; Cazzamali, Giuseppe

2015-04-01

Insulin-like growth factor binding proteins (IGFBPs) display many functions in humans including regulation of the insulin-like growth factor (IGF) signaling pathway. The various roles of human IGFBPs make them attractive protein candidates in drug discovery. Structural and functional knowledge on human proteins with therapeutic relevance is needed to design and process the next generation of protein therapeutics. In order to conduct structural and functional investigations large quantities of recombinant proteins are needed. However, finding a suitable recombinant production system for proteins such as full-length human IGFBPs, still remains a challenge. Here we present a mammalian HEK293 expression method suitable for over-expression of secretory full-length human IGFBP-1 to -7. Protein purification of full-length human IGFBP-1, -2, -3 and -5 was conducted using a two-step chromatography procedure and the final protein yields were between 1 and 12mg protein per liter culture media. The recombinant IGFBPs contained PTMs and exhibited high-affinity interactions with their natural ligands IGF-1 and IGF-2. Copyright © 2014 Elsevier Inc. All rights reserved.

[CNC proteins in physiology and pathology].

PubMed

Gęgotek, Agnieszka; Skrzydlewska, Elżbieta

2015-07-06

CNC proteins consist of Bach1, Bach2 and 4 homologous transcription factors: Nrf1, Nrf2, Nrf3 and p45NF-E2. Transcription factors belonging to this group of proteins play a crucial role in protection of cells against oxidative stress. Under physiological conditions, they remain in the cytoplasm in the inactive form or are degraded. However, in oxidative stress conditions, they are translocated to the nucleus, and bind to DNA in the ARE sequence. Consequently, there is transcription of genes encoding cytoprotective proteins, such as phase II enzymes, or low molecular weight antioxidant proteins (i.e., thioredoxin, ferritin, metallothionein) responsible for protecting cells from reactive oxygen species (ROS) action. The activity of transcriptional proteins depends directly on the redox state of the cell. ROS as second messenger signals, control inhibitors of cytoplasmic CNC proteins or potentiate the activity of kinases (MAPK, PKC, PI3K, PERK), leading to phosphorylation of transcription factors. This is conducive to translocation of these molecules into the nucleus and to formation of complexes that initiate the gene expression. Disorders of regulation of the activity of transcription factors belonging to the CNC proteins caused by gene mutations, epigenetic modifications or increased activity of p62, p21, or k-Ras, B-Raf and c-Myc oncogenes, induce changes in the level of ARE-dependent gene expression, which can lead even to the development of carcinogenesis. On the other hand, Nrf transcription factors, inducing the expression of antioxidants and enzymes responsible for the detoxification of xenobiotics, can be considered as a potential target of the action of chemopreventive factors in anticancer therapy.

Genetics Home Reference: anhidrotic ectodermal dysplasia with immune deficiency

MedlinePlus

... The proteins produced from these two genes regulate nuclear factor-kappa-B. Nuclear factor-kappa-B is a group of related ... proteins with impaired function, which reduces activation of nuclear factor-kappa-B. These changes disrupt certain signaling ...

Dissecting protein:protein interactions between transcription factors with an RNA aptamer.

PubMed Central

Tian, Y; Adya, N; Wagner, S; Giam, C Z; Green, M R; Ellington, A D

1995-01-01

Nucleic acid aptamers isolated from random sequence pools have generally proven useful at inhibiting the interactions of nucleic acid binding proteins with their cognate nucleic acids. In order to develop reagents that could also be used to study protein:protein interactions, we have used in vitro selection to search for RNA aptamers that could interact with the transactivating protein Tax from human T-cell leukemia virus. Tax does not normally bind to nucleic acids, but instead stimulates transcription by interacting with a variety of cellular transcription factors, including the cyclic AMP-response element binding protein (CREB), NF-kappa B, and the serum response factor (SRF). Starting from a pool of greater than 10(13) different RNAs with a core of 120 random sequence positions, RNAs were selected for their ability to be co-retained on nitrocellulose filters with Tax. After five cycles of selection and amplification, a single nucleic acid species remained. This aptamer was found to bind Tax with high affinity and specificity, and could disrupt complex formation between Tax and NF-kappa B, but not with SRF. The differential effects of our aptamer probe on protein:protein interactions suggest a model for how the transcription factor binding sites on the surface of the Tax protein are organized. This model is consistent with data from a variety of other studies. PMID:7489503

Preclinical and clinical studies on the use of growth factors for bone repair: a systematic review.

PubMed

Fisher, Daniel Mark; Wong, James Min-Leong; Crowley, Conor; Khan, Wasim S

2013-05-01

Bone healing is a complex process. Whilst the majority of fractures heal with conventional treatment, open fractures, large bone defects and non unions still provide great challenges to Orthopaedic Surgeons. Whilst autologous bone graft is seen as the gold standard, the use of growth factors is a growing area of research to find an effective alternative with lower side effects such as donor site morbidity and the finite amount available. This systematic review aims to summarize the pre clinical in-vivo studies and examine the clinical studies on the use of growth factors in bone healing. Databases: PubMed, Medline, OVID, and Cochrane library. The following key words and search terms were used: Growth Factors, Bone Healing, Bone Morphogenic Protein, Transforming Growth Factor Beta, Insulin Like Growth Factor, Platelet Derived Growth Factor, Fracture. All articles were screened based on title with abstracts and full text articles reviewed as appropriate. Reference lists were reviewed from relevant articles to ensure comprehensive and systematic review. Three tables of studies were constructed focussing on Bone Morphogenic Proteins, Platelet Rich Plasma and Growth Factors and Tissue Engineering. Bone Morphogenic Proteins and Platelet Rich Plasma, which contains multiple growth factors, have been shown in preclinical and clinical trials to be an effective alternative to autologous bone graft. Bone Morphogenic Proteins have been shown to be effective in fracture non union, and in open tibial fractures. Platelet Rich Plasma has shown promise in preclinical trials and some small clinical trials, however numbers are limited. Bone Morphogenic Proteins have been shown to be superior to Platelet Rich Protein in one trial. Combining these growth factors with tissue engineering techniques is the focus of ongoing research, and through further clinical trials the most effective techniques for enhancing bone healing will be revealed.

The ubiquitin ligase Nedd4 mediates oxidized low-density lipoprotein-induced downregulation of insulin-like growth factor-1 receptor

PubMed Central

Higashi, Yusuke; Sukhanov, Sergiy; Parthasarathy, Sampath; Delafontaine, Patrice

2008-01-01

Oxidized low-density lipoprotein (LDL) is proatherogenic and induces smooth muscle cell apoptosis, which contributes to atherosclerotic plaque destabilization. We showed previously that oxidized LDL downregulates insulin-like growth factor-1 receptor in human smooth muscle cells and that this is critical for induction of apoptosis. To identify mechanisms, we exposed smooth muscle cells to 60 μg/ml oxidized LDL or native LDL and assessed insulin-like growth factor-1 receptor mRNA levels, protein synthesis rate, and receptor protein stability. Oxidized LDL decreased insulin-like growth factor-1 receptor mRNA levels by 30% at 8 h compared with native LDL, and this decrease was maintained for up to 20 h. However, insulin-like growth factor-1 receptor protein synthesis rate was not altered by oxidized LDL. Pulse-chase labeling experiments revealed that oxidized LDL reduced insulin-like growth factor-1 receptor protein half-life to 12.2 ± 1.7 h from 24.4 ± 4.7 h with native LDL. This destabilization of insulin-like growth factor-1 receptor protein was accompanied by enhanced receptor ubiquitination. Overexpression of dominant-negative Nedd4 prevented oxidized LDL-induced downregulation of insulin-like growth factor-1 receptor, suggesting that Nedd4 was the ubiquitin ligase that mediated receptor downregulation. However, the proteasome inhibitors lactacystin, MG-132, and proteasome inhibitor-1 failed to block oxidized LDL-induced downregulation of insulin-like growth factor-1 receptor. Thus oxidized LDL downregulates insulin-like growth factor-1 receptor by destabilizing the protein via Nedd4-enhanced ubiquitination, leading to degradation via a proteasome-independent pathway. This finding provides novel insights into oxidized LDL-triggered oxidant signaling and mechanisms of smooth muscle cell depletion that contribute to plaque destabilization and coronary events. PMID:18723765

A high sensitivity MEA probe for measuring real time rat brain glucose flux.

PubMed

Wei, Wenjing; Song, Yilin; Shi, Wentao; Lin, Nansen; Jiang, Tingjun; Cai, Xinxia

2014-05-15

The mammalian central nervous system (CNS) relies on a constant supply of external glucose for its undisturbed operation. This article presents an implantable Multi-Electrode Array (MEA) probe for brain glucose measurement. The MEA was implemented on Silicon-On-Insulator (SOI) wafer using Micro-Electro-Mechanical-Systems (MEMS) methods. There were 16 platinum recording sites on the probe and enzyme glucose oxidase (GOx) was immobilized on them. The glucose sensitivity of the MEA probe was as high as 489 µA mM(-1) cm(-2). 1,3-Phenylenediamine (mPD) was electropolymerized onto the Pt recording surfaces to prevent larger molecules such as ascorbic acid (AA), 3,4-dihydroxyphenylacetic acid (DOPAC), serotonin (5-HT), and dopamine (DA) from reaching the recording sites surface. The MEA probe was implanted in the anesthetized rat striatum and responded to glucose levels which were altered by intraperitoneal injection of glucose and insulin. After the in vivo experiment, the MEA probe still kept sensitivity to glucose, these suggested that the MEA probe was reliable for glucose monitoring in brain extracellular fluid (ECF). © 2013 Published by Elsevier B.V.

Long-term studies on the economic impact of ticks on Sanga cattle in Zambia.

PubMed

de Castro, J J; James, A D; Minjauw, B; Di Giulio, G U; Permin, A; Pegram, R G; Chizyuka, H G; Sinyangwe, P

1997-01-01

Three different tick control policies were tested in groups of traditionally managed Sanga cattle in the Central Province of Zambia over a period of 3 years. One group was given strategic tick control using 12 pyrethroid acaricide spray applications between the onset and the end of the wet season (October to March). The productivity of this herd was compared with that of a group with no tick control and a group under an intensive tick control regimen of spraying every week in the wet season and every 2 weeks in the dry season (36 applications per year). The highest output was associated with intensive tick control, followed by strategic control and then no tick control policies. However, when the costs of tick control were taken into account, the strategic tick control policy produced the best economic result, followed by the intensive and then the no tick control policies. Neither the strategic nor the intensive tick control policy was sufficient to prevent the transmission of East Coast fever (ECF) infection when this disease was introduced to the area.

A Recommendation for Naming Transcription Factor Proteins in the Grasses

USDA-ARS?s Scientific Manuscript database

Transcription factors are central for the exquisite temporal and spatial expression patterns of many genes. These proteins are characterized by their ability to be tethered to particular regulatory sequences in the genes that they control. While many other proteins participate in the regulation of g...

The Relationship of Novel Plasma Proteins in the Early Neonatal Period With Retinopathy of Prematurity

PubMed Central

Lynch, Anne M.; Wagner, Brandie D.; Mandava, Naresh; Palestine, Alan G.; Mourani, Peter M.; McCourt, Emily A.; Oliver, Scott C. N.; Abman, Steven H.

2016-01-01

Purpose Retinopathy of prematurity (ROP) is a vision-threatening disease associated with abnormal retinal vascular development. Proteins from the insulin-like growth factor pathway are related to ROP. However, there is a paucity of research on the role of other proteins in ROP. The aim of this study was to identify plasma proteins related to clinically significant ROP. Methods We measured 1121 plasma proteins in the early neonatal period in infants at risk for ROP using an aptamer-based proteomic technology. The primary aim of the study was to compare plasma protein concentrations in infants who did (n = 12) and did not (n = 23) subsequently develop clinically significant ROP using logistic regression. As a secondary aim, we examined patterns in the proteins across categories of clinically significant, low-grade, and no ROP groups. Results Lower levels of 16 proteins were associated with an increased risk of clinically significant ROP. In this group, superoxide dismutase (Mn), mitochondrial (MnSOD), and chordin-like protein 1 (CRDL1) were highly ranked. Other proteins in this group included: C-C motif chemokine 14 (HCC-1), prolactin, insulin-like growth factor-binding protein 7 (IGFBP-7), and eotaxin. Higher levels of 12 proteins were associated with a higher risk for ROP. Fibroblast growth factor 19 (FGF-19) was the top-ranked protein target followed by hepatocyte growth factor-like protein (MSP), luteinizing hormone (LH), cystatin M, plasminogen, and proprotein convertase subtilisin/kexin type 9 (PCSK9). We also noted different patterns in the trend of concentrations of proteins across the clinically significant, low-grade, and no ROP groups. Conclusions We discovered plasma proteins with novel associations with clinically significant ROP (MnSOD, CRDL1, PCSK9), proteins with links to established ROP signaling pathways (IGFBP-7), and proteins such as MnSOD that may be a target for future therapeutic interventions. PMID:27679852

Plant Abiotic Stress Proteomics: The Major Factors Determining Alterations in Cellular Proteome

PubMed Central

Kosová, Klára; Vítámvás, Pavel; Urban, Milan O.; Prášil, Ilja T.; Renaut, Jenny

2018-01-01

HIGHLIGHTS: Major environmental and genetic factors determining stress-related protein abundance are discussed.Major aspects of protein biological function including protein isoforms and PTMs, cellular localization and protein interactions are discussed.Functional diversity of protein isoforms and PTMs is discussed. Abiotic stresses reveal profound impacts on plant proteomes including alterations in protein relative abundance, cellular localization, post-transcriptional and post-translational modifications (PTMs), protein interactions with other protein partners, and, finally, protein biological functions. The main aim of the present review is to discuss the major factors determining stress-related protein accumulation and their final biological functions. A dynamics of stress response including stress acclimation to altered ambient conditions and recovery after the stress treatment is discussed. The results of proteomic studies aimed at a comparison of stress response in plant genotypes differing in stress adaptability reveal constitutively enhanced levels of several stress-related proteins (protective proteins, chaperones, ROS scavenging- and detoxification-related enzymes) in the tolerant genotypes with respect to the susceptible ones. Tolerant genotypes can efficiently adjust energy metabolism to enhanced needs during stress acclimation. Stress tolerance vs. stress susceptibility are relative terms which can reflect different stress-coping strategies depending on the given stress treatment. The role of differential protein isoforms and PTMs with respect to their biological functions in different physiological constraints (cellular compartments and interacting partners) is discussed. The importance of protein functional studies following high-throughput proteome analyses is presented in a broader context of plant biology. In summary, the manuscript tries to provide an overview of the major factors which have to be considered when interpreting data from proteomic studies on stress-treated plants. PMID:29472941

Lewis lung carcinoma regulation of mechanical stretch-induced protein synthesis in cultured myotubes.

PubMed

Gao, Song; Carson, James A

2016-01-01

Mechanical stretch can activate muscle and myotube protein synthesis through mammalian target of rapamycin complex 1 (mTORC1) signaling. While it has been established that tumor-derived cachectic factors can induce myotube wasting, the effect of this catabolic environment on myotube mechanical signaling has not been determined. We investigated whether media containing cachectic factors derived from Lewis lung carcinoma (LLC) can regulate the stretch induction of myotube protein synthesis. C2C12 myotubes preincubated in control or LLC-derived media were chronically stretched. Protein synthesis regulation by anabolic and catabolic signaling was then examined. In the control condition, stretch increased mTORC1 activity and protein synthesis. The LLC treatment decreased basal mTORC1 activity and protein synthesis and attenuated the stretch induction of protein synthesis. LLC media increased STAT3 and AMP-activated protein kinase phosphorylation in myotubes, independent of stretch. Both stretch and LLC independently increased ERK1/2, p38, and NF-κB phosphorylation. In LLC-treated myotubes, the inhibition of ERK1/2 and p38 rescued the stretch induction of protein synthesis. Interestingly, either leukemia inhibitory factor or glycoprotein 130 antibody administration caused further inhibition of mTORC1 signaling and protein synthesis in stretched myotubes. AMP-activated protein kinase inhibition increased basal mTORC1 signaling activity and protein synthesis in LLC-treated myotubes, but did not restore the stretch induction of protein synthesis. These results demonstrate that LLC-derived cachectic factors can dissociate stretch-induced signaling from protein synthesis through ERK1/2 and p38 signaling, and that glycoprotein 130 signaling is associated with the basal stretch response in myotubes. Copyright © 2016 the American Physiological Society.

Refolding of soluble leukemia inhibitory factor receptor fusion protein (gp 190 sol DAF) from urea.

PubMed

Liu, H; Moreau, J F; Gualde, N; Fu, J

1997-04-01

The insoluble inclusion bodies of soluble leukemia inhibitory factor receptor fusion protein (gp 190 sol DAF) was solubilized in 8 M urea on the unfolding transitions, and several factors on the aggregate formation were indirectly analyzed for the refolding of gp 190 sol DAF. Results indicate that the refolding yield can be considerably increased at lowering concentration of the unfolding protein, a little soluble protein with the slow refolding appears in the process of the aggregate formation and the concentration of the denaturant must be down to a minimum level for its refolding.

Transcription Factor IIB (TFIIB)-Related Protein (pBrp), a Plant-Specific Member of the TFIIB-Related Protein Family

PubMed Central

Lagrange, Thierry; Hakimi, Mohamed-Ali; Pontier, Dominique; Courtois, Florence; Alcaraz, Jean Pierre; Grunwald, Didier; Lam, Eric; Lerbs-Mache, Silva

2003-01-01

Although it is now well documented that metazoans have evolved general transcription factor (GTF) variants to regulate their complex patterns of gene expression, there is so far no information regarding the existence of specific GTFs in plants. Here we report the characterization of a ubiquitously expressed gene that encodes a bona fide novel transcription factor IIB (TFIIB)-related protein in Arabidopsis thaliana. We have shown that this protein is the founding member of a plant-specific TFIIB-related protein family named pBrp (for plant-specific TFIIB-related protein). Surprisingly, in contrast to common GTFs that are localized in the nucleus, the bulk of pBrp proteins are bound to the cytoplasmic face of the plastid envelope, suggesting an organelle-specific function for this novel class of TFIIB-related protein. We show that pBrp proteins harbor conditional proteolytic signals that can target these proteins for rapid turnover by the proteasome-mediated protein degradation pathway. Interestingly, under conditions of proteasome inhibition, pBrp proteins accumulate in the nucleus. Together, our results suggest a possible involvement of these proteins in an intracellular signaling pathway between plastids and the nucleus. Our data provide the first evidence for an organelle-related evolution of the eukaryotic general transcription machinery. PMID:12697827

Enhancement of the efficacy of therapeutic proteins by formulation with PEGylated liposomes; a case of FVIII, FVIIa and G-CSF.

PubMed

Yatuv, Rivka; Robinson, Micah; Dayan, Inbal; Baru, Moshe

2010-02-01

Improving the pharmacodynamics of protein drugs has the potential to improve the care and the quality of life of patients suffering from a variety of diseases. Four approaches to improve protein drugs are described: PEGylation, amino acid substitution, fusion to carrier proteins and encapsulation. A new platform technology based on the binding of proteins/peptides to the outer surface of PEGylated liposomes (PEGLip) is then presented. Binding of proteins to PEGLip is non-covalent, highly specific and dependent on an amino acid consensus sequence within the proteins. Association of proteins with PEGLip results in substantial enhancement of the pharmacodynamic properties of proteins following administration. This has been demonstrated in preclinical studies and clinical trials with coagulation factors VIII and VIIa. It has also been demonstrated in preclinical studies with granulocyte colony-stimulating factor. A mechanism is presented that explains the improvements in hemostatic efficacy of PEGLip-formulated coagulation factors VIII and VIIa. The reader will gain an understanding of the advantages and disadvantages of each of the approaches discussed. PEGLip formulation is an important new approach to improve the pharmacodynamics of protein drugs. This approach may be applied to further therapeutic proteins in the future.

Investigation of carbohydrate and protein metabolism in the digestive organs of the rabbit under the combined influence of vibration, acceleration and irradiation

NASA Technical Reports Server (NTRS)

Yuy, R. I.

1975-01-01

During spaceflight, the organism is subjected to the influence of various extremal factors such as acceleration, vibration, irradiation, etc. The study of the influence of these factors on metabolism, especially carbohydrate and protein metabolism, in young rabbits is of great significance in simulation experiments. Dynamic factors and irradiation, depending on dose and duration, lead to reduced RNA and protein metabolism.

Identification of pathogenic factors potentially involved in Staphylococcus aureus keratitis using proteomics.

PubMed

Khan, Shamila; Cole, Nerida; Hume, Emma B H; Garthwaite, Linda L; Nguyen-Khuong, Terry; Walsh, Bradley J; Willcox, Mark D P

2016-10-01

Staphylococcus is a leading cause of microbial keratitis, characterized by destruction of the cornea by bacterial exoproteins and host-associated factors. The aim of this study was to compare extracellular and cell-associated proteins produced by two different isolates of S. aureus, a virulent clinical isolate (Staph 38) and a laboratory strain (Staphylococcus aureus 8325-4) of weaker virulence in the mouse keratitis model. Proteins were analyzed using 2D polyacrylamide gel electrophoresis and identified by subsequent mass spectrometry. Activity of staphylococcal adhesins was assessed by allowing strains to bind to various proteins adsorbed onto polymethylmethacrylate squares. Thirteen proteins in the extracellular fraction and eight proteins in the cell-associated fractions after bacterial growth were produced in increased amounts in the clinical isolate Staph 38. Four of these proteins were S. aureus virulence factor adhesins, fibronectin binding protein A, staphopain, glyceraldehyde-3-phosphate dehydrogenase 2 and extracellular adherence protein. The clinical isolate Staph 38 adhered to a greater extent to all mammalian proteins tested, indicating the potential of the adhesins to be active on its surface. Other proteins with increased expression in Staph 38 included potential moonlighting proteins and proteins involved in transcription or translation. This is the first demonstration of the proteome of S. aureus isolates from keratitis. These results indicate that the virulent clinical isolate produces more potentially important virulence factors compared to the less virulent laboratory strain and these may be associated with the ability of a S. aureus strain to cause more severe keratitis. Copyright © 2016 Elsevier Ltd. All rights reserved.

Universal features of fluctuations in globular proteins.

PubMed

Erman, Burak

2016-06-01

Using data from 2000 non-homologous protein crystal structures, we show that the distribution of residue B factors of proteins collapses onto a single master curve. We show by maximum entropy arguments that this curve is a Gamma function whose order and dispersion are obtained from experimental data. The distribution for any given specific protein can be generated from the master curve by a linear transformation. Any perturbation of the B factor distribution of a protein, imposed at constant energy, causes a decrease in the entropy of the protein relative to that of the reference state. Proteins 2016; 84:721-725. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Sperm and Spermatids Contain Different Proteins and Bind Distinct Egg Factors

PubMed Central

Teperek, Marta; Miyamoto, Kei; Simeone, Angela; Feret, Renata; Deery, Michael J.; Gurdon, John B.; Jullien, Jerome

2014-01-01

Spermatozoa are more efficient at supporting normal embryonic development than spermatids, their immature, immediate precursors. This suggests that the sperm acquires the ability to support embryonic development during spermiogenesis (spermatid to sperm maturation). Here, using Xenopus laevis as a model organism, we performed 2-D Fluorescence Difference Gel Electrophoresis (2D-DIGE) and mass spectrometry analysis of differentially expressed proteins between sperm and spermatids in order to identify factors that could be responsible for the efficiency of the sperm to support embryonic development. Furthermore, benefiting from the availability of egg extracts in Xenopus, we also tested whether the chromatin of sperm could attract different egg factors compared to the chromatin of spermatids. Our analysis identified: (1) several proteins which were present exclusively in sperm; but not in spermatid nuclei and (2) numerous egg proteins binding to the sperm (but not to the spermatid chromatin) after incubation in egg extracts. Amongst these factors we identified many chromatin-associated proteins and transcriptional repressors. Presence of transcriptional repressors binding specifically to sperm chromatin could suggest its preparation for the early embryonic cell cycles, during which no transcription is observed and suggests that sperm chromatin has a unique protein composition, which facilitates the recruitment of egg chromatin remodelling factors. It is therefore likely that the acquisition of these sperm-specific factors during spermiogenesis makes the sperm chromatin suitable to interact with the maternal factors and, as a consequence, to support efficient embryonic development. PMID:25244019

[Chlamydia trachomatis proteasome protein as one of the significant pathogenicity factors of exciter].

PubMed

Davydov, D Iu; Zigangirova, N A

2014-01-01

Sex-related infections are a global problem. Such infections may lead to acute or chronic diseases. Chlamydia trachomatis is a dangerous and widespread pathogenicity factor that is not sensitive to conventional drugs and has no obvious symptoms. Protein CPAF is leading factor of pathogenesis. This protein inhibits the signaling pathways of host cell and supports long survival of the pathogen in the host cell. The goal of this work was to review general properties of the proteasome Chlamydia protein CPAF, its functions, and role in pathology. The role of protein CPAF in the anti-chlamydia immune reaction is discussed. The prospects of the development of promising anti-chlamydia vaccine, as well as new effective anti-chlamydia drugs are also discussed.

Myeloid Leukemia Factor Acts in a Chaperone Complex to Regulate Transcription Factor Stability and Gene Expression.

PubMed

Dyer, Jamie O; Dutta, Arnob; Gogol, Madelaine; Weake, Vikki M; Dialynas, George; Wu, Xilan; Seidel, Christopher; Zhang, Ying; Florens, Laurence; Washburn, Michael P; Abmayr, Susan M; Workman, Jerry L

2017-06-30

Mutations that affect myelodysplasia/myeloid leukemia factor (MLF) proteins are associated with leukemia and several other cancers. However, with no strong homology to other proteins of known function, the role of MLF proteins in the cell has remained elusive. Here, we describe a proteomics approach that identifies MLF as a member of a nuclear chaperone complex containing a DnaJ protein, BCL2-associated anthanogene 2, and Hsc70. This complex associates with chromatin and regulates the expression of target genes. The MLF complex is bound to sites of nucleosome depletion and sites containing active chromatin marks (e.g., H3K4me3 and H3K4me1). Hence, MLF binding is enriched at promoters and enhancers. Additionally, the MLF-chaperone complex functions to regulate transcription factor stability, including the RUNX transcription factor involved in hematopoiesis. Although Hsc70 and other co-chaperones have been shown to play a role in nuclear translocation of a variety of proteins including transcription factors, our findings suggest that MLF and the associated co-chaperones play a direct role in modulating gene transcription. Copyright © 2016 Elsevier Ltd. All rights reserved.

Interaction of Leptospira Elongation Factor Tu with Plasminogen and Complement Factor H: A Metabolic Leptospiral Protein with Moonlighting Activities

PubMed Central

Abe, Cecília M.; Monaris, Denize; Morais, Zenaide M.; Souza, Gisele O.; Vasconcellos, Sílvio A.; Isaac, Lourdes; Abreu, Patrícia A. E.; Barbosa, Angela S.

2013-01-01

The elongation factor Tu (EF-Tu), an abundant bacterial protein involved in protein synthesis, has been shown to display moonlighting activities. Known to perform more than one function at different times or in different places, it is found in several subcellular locations in a single organism, and may serve as a virulence factor in a range of important human pathogens. Here we demonstrate that Leptospira EF-Tu is surface-exposed and performs additional roles as a cell-surface receptor for host plasma proteins. It binds plasminogen in a dose-dependent manner, and lysine residues are critical for this interaction. Bound plasminogen is converted to active plasmin, which, in turn, is able to cleave the natural substrates C3b and fibrinogen. Leptospira EF-Tu also acquires the complement regulator Factor H (FH). FH bound to immobilized EF-Tu displays cofactor activity, mediating C3b degradation by Factor I (FI). In this manner, EF-Tu may contribute to leptospiral tissue invasion and complement inactivation. To our knowledge, this is the first description of a leptospiral protein exhibiting moonlighting activities. PMID:24312361

Acquired activated protein C resistance associated with anti-protein S antibody as a strong risk factor for DVT in non-SLE patients.

PubMed

Nojima, Junzo; Kuratsune, Hirohiko; Suehisa, Etsuji; Kawasaki, Tomio; Machii, Takashi; Kitani, Teruo; Iwatani, Yoshinori; Kanakura, Yuzuru

2002-11-01

Anti-phospholipid (aPL) antibodies (Abs) are well known to be associated with thromboembolic events in patients with systemic lupus erythematosus (SLE). However, the clinical relevance of a PL Abs in patients without SLE (non-SLE) who have venous thromboembolism remains unclear. We evaluated 143 non-SLE patients with a first episode of clinically suspected deep vein thrombosis (DVT) by using objective tests for diagnosing DVT and laboratory tests including the activated protein C resistance (APC-R) test, the factor V Leiden test, and various aPL Abs. The prevalence of acquired APC-R, in which case there was no factor V Leiden mutation, was significantly higher in patients with DVT (15/58 cases, 25.9%, p < 0.0001) than in those without DVT (3/80 cases, 3.7%), and confirmed that acquired APC-R was a strong risk factor for DVT (odds ratio [OR], 8.95; 95% confidence intervals [CI], 2.45-32.7; p < 0.001). Multivariate logistic analysis revealed that the presence of LA, aCL, anti-beta2-glycoprotein I, anti-prothrombin and anti-protein C Abs was not reliable as a risk factor for DVT in non-SLE patients, and that the presence of anti-protein S Abs was the most significant risk factor for DVT (OR, 5.88; 95% CI, 1.96-17.7; p < 0.002). Furthermore, the presence of anti-protein S Abs was strongly associated with acquired APC-R (OR, 57.8; 95% CI, 8.53-391; p < 0.0001). These results suggest that acquired APC-R may reflect functional interference by anti-protein S Abs of the protein C pathway, which action may represent an important mechanism for the development DVT in non-SLE patients.

What we know about ST13, a co-factor of heat shock protein, or a tumor suppressor?*

PubMed Central

Shi, Zheng-zheng; Zhang, Jia-wei; Zheng, Shu

2007-01-01

This article is to summarize the molecular and functional analysis of the gene “suppression of tumorigenicity 13” (ST13). ST13 is in fact the gene encoding Hsp70 interacting protein (Hip), a co-factor (co-chaperone) of the 70-kDa heat shock proteins (Hsc/Hsp70). By collaborating with other positive co-factors such as Hsp40 and the Hsp70-Hsp90 organizing protein (Hop), or competing with negative co-factors such as Bcl2-associated athanogen 1 (Bag1), Hip may facilitate the chaperone function of Hsc/Hsp70 in protein folding and repair, and in controlling the activity of regulatory proteins such as steroid receptors and regulators of proliferation or apoptosis. Although the nomenclature of ST13 implies a role in the suppression of tumorigenicity (ST), to date available experimental data are not sufficient to support its role in cancer development, except for the possible down-regulation of ST13 in gastric and colorectal cancers. Further investigation of this gene at the physiological level would benefit our understanding of diseases such as endocrinological disorders, cancer, and neurodegeneration commonly associated with protein misfolding. PMID:17323428

Analyzing the Effects of Climate Factors on Soybean Protein, Oil Contents, and Composition by Extensive and High-Density Sampling in China.

PubMed

Song, Wenwen; Yang, Ruping; Wu, Tingting; Wu, Cunxiang; Sun, Shi; Zhang, Shouwei; Jiang, Bingjun; Tian, Shiyan; Liu, Xiaobing; Han, Tianfu

2016-05-25

From 2010 to 2013, 763 soybean samples were collected from an extensive area of China. The correlations between seed compositions and climate data were analyzed. The contents of crude protein and water-soluble protein, total amount of protein plus oil, and most of the amino acids were positively correlated with an accumulated temperature ≥15 °C (AT15) and the mean daily temperature (MDT) but were negatively correlated with hours of sunshine (HS) and diurnal temperature range (DTR). The correlations of crude oil and most fatty acids with climate factors were opposite to those of crude protein. Crude oil content had a quadratic regression relationship with MDT, and a positive correlation between oil content and MDT was found when the daily temperature was <19.7 °C. A path analysis indicated that DTR was the main factor that directly affected soybean protein and oil contents. The study illustrated the effects of climate factors on soybean protein and oil contents and proposed agronomic practices for improving soybean quality in different regions of China. The results provide a foundation for the regionalization of high-quality soybean production in China and similar regions in the world.

Immunoproteomic identification of immunogenic proteins in Cronobacter sakazakii strain BAA-894.

PubMed

Wang, Jian; Du, Xin-Jun; Lu, Xiao-Nan; Wang, Shuo

2013-03-01

Cronobacter spp. are emerging opportunistic pathogens. Cronobacter sakazakii is considered as the predominant species in all infections. So far, our understanding of the species' immunogens and potential virulence factors of Cronobacter spp. remains limited. In this study, an immunoproteomic approach was used to investigate soluble and insoluble proteins from the genome-sequenced strain C. sakazakii ATCC BAA-894. Proteins were separated using two-dimensional electrophoresis, detected by Western blotting with polyclonal antibodies of C. sakazakii BAA-894, and identified using tandem mass spectrometry (MALDI-MS and MALDI-MS/MS, MS/MSMS). A total of 11 immunoreactive proteins were initially identified in C. sakazakii BAA-894, including two outer membrane proteins, four periplasmic proteins, and five cytoplasmic proteins. In silico functional analysis of the 11 identified proteins indicated three proteins that were initially described as immunogens of pathogenic bacteria. For the remaining eight proteins, one protein was categorized as a potential virulence factor involved in protection against reactive oxygen species, and seven proteins were considered to play potential roles in adhesion, invasion, and biofilm formation. To our knowledge, this is the first time that immunogenic proteins of C. sakazakii BAA-894 have been identified as immunogens and potential virulence factors by an immunoproteomics approach. Future studies should investigate the roles of these proteins in bacterial pathogenesis and modulation of host immune responses during infection to identify their potential as molecular therapeutic targets.

Quality Matters: Extension of Clusters of Residues with Good Hydrophobic Contacts Stabilize (Hyper)Thermophilic Proteins

PubMed Central

2015-01-01

Identifying determinant(s) of protein thermostability is key for rational and data-driven protein engineering. By analyzing more than 130 pairs of mesophilic/(hyper)thermophilic proteins, we identified the quality (residue-wise energy) of hydrophobic interactions as a key factor for protein thermostability. This distinguishes our study from previous ones that investigated predominantly structural determinants. Considering this key factor, we successfully discriminated between pairs of mesophilic/(hyper)thermophilic proteins (discrimination accuracy: ∼80%) and searched for structural weak spots in E. coli dihydrofolate reductase (classification accuracy: 70%). PMID:24437522

Isomer profiling of perfluorinated substances as a tool for source tracking: a review of early findings and future applications.

PubMed

Benskin, Jonathan P; De Silva, Amila O; Martin, Jonathan W

2010-01-01

The two major manufacturing techniques for perfluorochemicals can be distinguished based on the isomeric profile of their products. ECF (major use from 1950s to 2002) results in a product containing both linear and branched isomers, while telomerization (major use from 2002 to present) typically yields an isomerically pure, linear product. Among the most important question today, which has implication for future regulation of these chemicals, is to what extent human and environmental exposure is from historical products (i.e., ECF) versus currently manufactured fluorochemicals (i.e., telomer). Perfluoroalkyl-chain branching can also affect the physical and chemical properties of these chemicals, which may influence their environmental transport and degradation, partitioning, bioaccumulation, pharmacokinetics, and toxicity. Unless perfluorinated substances are considered as individual isomers, much of this information will be overlooked or missed altogether, which could potentially lead to inaccuracies in human and environmental risk assessments. In this review, we have highlighted novel findings, current knowledge gaps, and areas for improvement based on early experiments on the disposition of PFA and PFA-precursor isomers in the environment. We have also emphasized the wealth of information that can potentially be gleaned from future work in this area, which renders routine adoption of isomer-specific methodologies an attractive and logical next step in the progression of fluorochemicals analysis. However, despite vast improvements in recent years, a fast and comprehensive method capable of separating all major PFA and PFA-precursor isomers, while removing interferences is still required before these methods becomes routine in most labs. Purified and characterized standards of PFOA and PFOS that have isomer profiles consistent with those of historically produced (i.e., 3M) PFOS and PFOA are also required. The limited data available on PFA isomer profiles that exist in the environment and the biological properties of each isomer suggest that examination of isomer profiles may yield clues on the source of PFA contamination to human and the environment. For example, contributions from historical versus current PFOA emissions can be quantified by examining the isomer profile in abiotic samples . Similarly, residual PFOS/PFOA in pre-2002 consumer products may be distinguished from directly emitted PFOS/PFOA by the existence of slight difference in isomer profile. PFOS signatures may also have the potential to distinguish between indirect exposure (via precursors) versus direct exposure (via the sulfonate), based on findings of isomer-specific and/or enantiospecific biotransformation in vitro. Isomer-specific monitoring extended to longer-chain PFAs may also be informative in determining current and historical exposure sources. Finally, given the recent increase of production of PFOSF-based chemicals, following their 2002 phase out, the ability of using isomer profiles to distinguish between historical and currently produced PFOS may also be possible.

Effects of vernal equinox solar eclipse on temperature and wind direction in Switzerland

NASA Astrophysics Data System (ADS)

Eugster, Werner; Emmel, Carmen; Wolf, Sebastian; Buchmann, Nina; McFadden, Joseph P.; Whiteman, Charles David

2017-12-01

The vernal equinox total solar eclipse of 20 March 2015 produced a maximum occultation of 65.8-70.1 % over Switzerland during the morning hours (09:22 to 11:48 CET). Skies were generally clear over the Swiss Alps due to a persistent high-pressure band between the UK and Russia associated with a rather weak pressure gradient over the continent. To assess the effects of penumbral shading on near-surface meteorology across Switzerland, air temperature data measured at 10 min intervals at 184 MeteoSwiss weather stations were used. Wind speed and direction data were available from 165 of these stations. Additionally, six Swiss FluxNet eddy covariance flux (ECF) sites provided turbulent measurements at 20 Hz resolution. During maximum occultation, the temperature drop was up to 5.8 K at a mountain site where cold air can pool in a topographic depression. The bootstrapped average of the maximum temperature drops of all 184 MeteoSwiss sites during the solar eclipse was 1.51 ± 0.02 K (mean ± SE). A detailed comparison with literature values since 1834 showed a temperature decrease of 2.6 ± 1.7 K (average of all reports), with extreme values up to 11 K. On fair weather days under weak larger-scale pressure gradients, local thermo-topographic wind systems develop that are driven by small-scale pressure and temperature gradients. At one ECF site, the penumbral shading delayed the morning transition from down-valley to up-valley wind conditions. At another site, it prevented this transition from occurring at all. Data from the 165 MeteoSwiss sites measuring wind direction did not show a consistent pattern of wind direction response to the passing of the penumbral shadow. These results suggest that the local topographic setting had an important influence on the temperature drop and the wind flow patterns during the eclipse. A significant cyclonic effect of the passing penumbral shadow was found in the elevation range ≈ 1700-2700 m a. s. l., but not at lower elevations of the Swiss Plateau. This contrasts with an earlier theory that the anticyclonic outflow should reach as far as ≈ 2400 km from the center of the eclipse, which would have included all of Switzerland during the 2015 eclipse. Thus, measurable effects of penumbral shading on the local wind system could be even found at ≈ 2000 km from the path of the eclipse (that is, Switzerland during the 2015 eclipse), and our results tend to lend support to a newer theory that the anticyclonic cold-air outflow from the center of the eclipse only extends ≈ 1600 km outwards, with cyclonic flow beyond that distance.

Two CGTCA motifs and a GHF1/Pit1 binding site mediate cAMP-dependent protein kinase A regulation of human growth hormone gene expression in rat anterior pituitary GC cells.

PubMed

Shepard, A R; Zhang, W; Eberhardt, N L

1994-01-21

We established the cis-acting elements which mediate cAMP responsiveness of the human growth hormone (hGH) gene in transiently transfected rat anterior pituitary tumor GC cells. Analysis of the intact hGH gene or hGH 5'-flanking DNA (5'-FR) coupled to the hGh cDNA or chloramphenicol acetyltransferase or luciferase genes, indicated that cAMP primarily stimulated hGH promoter activity. Cotransfection of a protein kinase A inhibitory protein cDNA demonstrated that the cAMP response was mediated by protein kinase A. Mutational analysis of the hGH promoter identified two core cAMP response element motifs (CGTCA) located at nucleotides -187/-183 (distal cAMP response element; dCRE) and -99/-95 (proximal cAMP response element; pCRE) and a pituitary-specific transcription factor (GHF1/Pit1) binding site at nucleotides -123/-112 (dGHF1) which were required for cAMP responsiveness. GHF1 was not a limiting factor, since overexpression of GHF1 in cotransfections increased basal but not forskolin induction levels. Gel shift analyses indicated that similar, ubiquitous, thermostable protein(s) specifically bound the pCRE and dCRE motifs. The CGTCA motif-binding factors were cAMP response element binding protein (CREB)/activating transcription factor-1 (ATF-1)-related, since the DNA-protein complex was competed by unlabeled CREB consensus oligonucleotide, specifically supershifted by antisera to CREB and ATF-1 but not ATF-2, and was bound by purified CREB with the same relative binding affinity (pCRE < dCRE < CREB) and mobility as the GC nuclear extract. UV cross-linking and Southwestern blot analyses revealed multiple DNA-protein interactions of which approximately 100- and approximately 45-kDa proteins were predominant; the approximately 45-kDa protein may represent CREB. These results indicate that CREB/ATF-1-related factors act coordinately with the cell-specific factor GHF1 to mediate cAMP-dependent regulation of hGH-1 gene transcription in anterior pituitary somatotrophs.

A Systematic Analysis of Factors Localized to Damaged Chromatin Reveals PARP-Dependent Recruitment of Transcription Factors.

PubMed

Izhar, Lior; Adamson, Britt; Ciccia, Alberto; Lewis, Jedd; Pontano-Vaites, Laura; Leng, Yumei; Liang, Anthony C; Westbrook, Thomas F; Harper, J Wade; Elledge, Stephen J

2015-06-09

Localization to sites of DNA damage is a hallmark of DNA damage response (DDR) proteins. To identify DDR factors, we screened epitope-tagged proteins for localization to sites of chromatin damaged by UV laser microirradiation and found >120 proteins that localize to damaged chromatin. These include the BAF tumor suppressor complex and the amyotrophic lateral sclerosis (ALS) candidate protein TAF15. TAF15 contains multiple domains that bind damaged chromatin in a poly-(ADP-ribose) polymerase (PARP)-dependent manner, suggesting a possible role as glue that tethers multiple PAR chains together. Many positives were transcription factors; > 70% of randomly tested transcription factors localized to sites of DNA damage, and of these, ∼90% were PARP dependent for localization. Mutational analyses showed that localization to damaged chromatin is DNA-binding-domain dependent. By examining Hoechst staining patterns at damage sites, we see evidence of chromatin decompaction that is PARP dependent. We propose that PARP-regulated chromatin remodeling at sites of damage allows transient accessibility of DNA-binding proteins. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

Pharmacological targeting of the transcription factor SOX18 delays breast cancer in mice

PubMed Central

Overman, Jeroen; Fontaine, Frank; Moustaqil, Mehdi; Mittal, Deepak; Sierecki, Emma; Sacilotto, Natalia; Zuegg, Johannes; Robertson, Avril AB; Holmes, Kelly; Salim, Angela A; Mamidyala, Sreeman; Butler, Mark S; Robinson, Ashley S; Lesieur, Emmanuelle; Johnston, Wayne; Alexandrov, Kirill; Black, Brian L; Hogan, Benjamin M; De Val, Sarah; Capon, Robert J; Carroll, Jason S; Bailey, Timothy L; Koopman, Peter; Jauch, Ralf; Smyth, Mark J; Cooper, Matthew A; Gambin, Yann; Francois, Mathias

2017-01-01

Pharmacological targeting of transcription factors holds great promise for the development of new therapeutics, but strategies based on blockade of DNA binding, nuclear shuttling, or individual protein partner recruitment have yielded limited success to date. Transcription factors typically engage in complex interaction networks, likely masking the effects of specifically inhibiting single protein-protein interactions. Here, we used a combination of genomic, proteomic and biophysical methods to discover a suite of protein-protein interactions involving the SOX18 transcription factor, a known regulator of vascular development and disease. We describe a small-molecule that is able to disrupt a discrete subset of SOX18-dependent interactions. This compound selectively suppressed SOX18 transcriptional outputs in vitro and interfered with vascular development in zebrafish larvae. In a mouse pre-clinical model of breast cancer, treatment with this inhibitor significantly improved survival by reducing tumour vascular density and metastatic spread. Our studies validate an interactome-based molecular strategy to interfere with transcription factor activity, for the development of novel disease therapeutics. DOI: http://dx.doi.org/10.7554/eLife.21221.001 PMID:28137359

Structural definition of a potent macrophage activating factor derived from vitamin D3-binding protein with adjuvant activity for antibody production.

PubMed

Yamamoto, N

1996-10-01

Incubation of human vitamin D3-binding protein (Gc protein), with a mixture of immobilized beta-galactosidase and sialidase, efficiently generated a potent macrophage activating factor, a protein with N-acetylgalactosamine as the remaining sugar. Stepwise incubation of Gc protein with immobilized beta-galactosidase and sialidase, and isolation of the intermediates with immobilized lectins, revealed that either sequence of hydrolysis of Gc glycoprotein by these glycosidases yields the macrophage-activating factor, implying that Gc protein carries a trisaccharide composed of N-acetylgalactosamine and dibranched galactose and sialic acid termini. A 3 hr incubation of mouse peritoneal macrophages with picomolar amounts of the enzymatically generated macrophage-activating factor (GcMAF) resulted in a greatly enhanced phagocytic activity. Administration of a minute amount (10-50 pg/mouse) of GcMAF resulted in a seven- to nine-fold enhanced phagocytic activity of macrophages. Injection of sheep red blood cells (SRBC) along with GcMAF into mice produced a large number of anti-SRBC antibody secreting splenic cells in 2-4 days.

[Transcription factors NF-kB, HIF-1, HIF-2, growth factor VEGF, VEGFR2 and carboanhydrase IX mRNA and protein level in the development of kidney cancer metastasis].

PubMed

Spirina, L V; Usynin, Y A; Yurmazov, Z A; Slonimskaya, E M; Kolegova, E S; Kondakova, I V

2017-01-01

Here, we have investigated the participation of nuclear factors NF-kB, HIF-1 and HIF-2, VEGF, VEGFR2, and carboanhydrase IX in clear-cell renal cancer. We have determined the expression and protein level of transcription factors, VEGF, VEGFR2, and carboanhydrase IX in tumor and normal tissues of 30 patients with kidney cancer. The Real-Time PCR and ELISA were used in the study. The low levels of HIF-1 mRNA expression associated with high levels of HIF-1 protein were also associated with metastasis. The expression levels of VEGF, VEGFR2, and their protein levels are increased in primary tumors of patients with disseminated kidney cancer compared to nonmetastatic cancer. No correlation was revealed between the content of mRNA and encoded proteins in the kidney cancer tissues. The changes in the ratios of mRNA levels and the respective proteins (HIF-1α, HIF-2, NF-kB, VEGF, VEGFR2, and carboanhydrase IX) may contribute to kidney-cancer metastasis.

Protein aggregation studied by forward light scattering and light transmission analysis

NASA Astrophysics Data System (ADS)

Penzkofer, A.; Shirdel, J.; Zirak, P.; Breitkreuz, H.; Wolf, E.

2007-12-01

The aggregation of the circadian blue-light photo-receptor cryptochrome from Drosophila melanogaster (dCry) is studied by transmission and forward light scattering measurement in the protein transparent wavelength region. The light scattering in forward direction is caused by Rayleigh scattering which is proportional to the degree of aggregation. The light transmission through the samples in the transparent region is reduced by Mie light scattering in all directions. It depends on the degree of aggregation and the monomer volume fill factor of the aggregates (less total scattering with decreasing monomer volume fill factor of protein globule) allowing a distinction between tightly packed protein aggregation (monomer volume fill factor 1) and loosely packed protein aggregation (monomer volume fill factor less than 1). An increase in aggregation with temperature, concentration, and blue-light exposure is observed. At a temperature of 4 °C and a protein concentration of less than 0.135 mM no dCry aggregation was observed, while at 24 °C and 0.327 mM gelation occurred (loosely packed aggregates occupying the whole solution volume).

Angiogenic therapy for cardiac repair based on protein delivery systems.

PubMed

Formiga, F R; Tamayo, E; Simón-Yarza, T; Pelacho, B; Prósper, F; Blanco-Prieto, M J

2012-05-01

Cardiovascular diseases remain the first cause of morbidity and mortality in the developed countries and are a major problem not only in the western nations but also in developing countries. Current standard approaches for treating patients with ischemic heart disease include angioplasty or bypass surgery. However, a large number of patients cannot be treated using these procedures. Novel curative approaches under investigation include gene, cell, and protein therapy. This review focuses on potential growth factors for cardiac repair. The role of these growth factors in the angiogenic process and the therapeutic implications are reviewed. Issues including aspects of growth factor delivery are presented in relation to protein stability, dosage, routes, and safety matters. Finally, different approaches for controlled growth factor delivery are discussed as novel protein delivery platforms for cardiac regeneration.

CHEMOSENSITIZATION BY A NON-APOPTOGENIC HEAT SHOCK PROTEIN 70-BINDING APOPTOSIS INDUCING FACTOR MUTANT

EPA Science Inventory

Chemosensitization by a non-apoptogenic heat shock protein 70-binding apoptosis inducing factor mutant

Abstract
HSP70 inhibits apoptosis by neutralizing the caspase activator Apaf-1 and by interacting with apoptosis inducing factor (AIF), a mitochondrial flavoprotein wh...

Hypoxia in Invasion and Metastasis

DTIC Science & Technology

2007-08-01

hypoxia and activating HIF-1 downregulate the DNA mismatch repair proteins ( mlh1 and/or msh2), a group of important proteins for maintaining genetic...Investigate the hypoxia and activating HIF-1 downregulate the DNA mismatch repair proteins ( mlh1 and/or msh2) (Month 7-12) Methods: We performed a parallel...inducible factors from invasive tumor cells. Changes in the level of multiple hypoxia related factor (HIF-1) and DNA mismatch repair proteins ( MLH1 , MSH2

Statistical approaches to maximize recombinant protein expression in Escherichia coli: a general review.

PubMed

Papaneophytou, Christos P; Kontopidis, George

2014-02-01

The supply of many valuable proteins that have potential clinical or industrial use is often limited by their low natural availability. With the modern advances in genomics, proteomics and bioinformatics, the number of proteins being produced using recombinant techniques is exponentially increasing and seems to guarantee an unlimited supply of recombinant proteins. The demand of recombinant proteins has increased as more applications in several fields become a commercial reality. Escherichia coli (E. coli) is the most widely used expression system for the production of recombinant proteins for structural and functional studies. However, producing soluble proteins in E. coli is still a major bottleneck for structural biology projects. One of the most challenging steps in any structural biology project is predicting which protein or protein fragment will express solubly and purify for crystallographic studies. The production of soluble and active proteins is influenced by several factors including expression host, fusion tag, induction temperature and time. Statistical designed experiments are gaining success in the production of recombinant protein because they provide information on variable interactions that escape the "one-factor-at-a-time" method. Here, we review the most important factors affecting the production of recombinant proteins in a soluble form. Moreover, we provide information about how the statistical design experiments can increase protein yield and purity as well as find conditions for crystal growth. Copyright © 2013 Elsevier Inc. All rights reserved.

Enteral delivery of proteins enhances the expression of proteins involved in the cytoskeleton and protein biosynthesis in human duodenal mucosa.

PubMed

Goichon, Alexis; Bertrand, Julien; Chan, Philippe; Lecleire, Stéphane; Coquard, Aude; Cailleux, Anne-Françoise; Vaudry, David; Déchelotte, Pierre; Coëffier, Moïse

2015-08-01

Amino acids are well known to be key effectors of gut protein turnover. We recently reported that enteral delivery of proteins markedly stimulated global duodenal protein synthesis in carbohydrate-fed healthy humans, but specifically affected proteins remain unknown. We aimed to assess the influence of an enteral protein supply on the duodenal mucosal proteome in carbohydrate-fed humans. Six healthy volunteers received for 5 h, on 2 occasions and in random order, either an enteral infusion of maltodextrins alone (0.25 g · kg⁻¹ · h⁻¹) mimicking the fed state or maltodextrins with a protein powder (0.14 g proteins · kg⁻¹ · h⁻¹). Endoscopic duodenal biopsy specimens were then collected and frozen until analysis. A 2-dimensional polyacrylamide gel electrophoresis-based comparative proteomics analysis was then performed, and differentially expressed proteins (at least ±1.5-fold change; Student's t test, P < 0.05) were identified by mass spectrometry. Protein expression changes were confirmed by Western blot analysis. Thirty-two protein spots were differentially expressed after protein delivery compared with maltodextrins alone: 28 and 4 spots were up- or downregulated, respectively. Among the 22 identified proteins, 11 upregulated proteins were involved either in the cytoskeleton (ezrin, moesin, plastin 1, lamin B1, vimentin, and β-actin) or in protein biosynthesis (glutamyl-prolyl-transfer RNA synthetase, glutaminyl-transfer RNA synthetase, elongation factor 2, elongation factor 1δ, and eukaryotic translation and initiation factor 3 subunit f). Enteral delivery of proteins altered the duodenal mucosal proteome and mainly stimulated the expression of proteins involved in cytoskeleton and protein biosynthesis. These results suggest that protein supply may affect intestinal morphology by stimulating actin cytoskeleton remodeling. © 2015 American Society for Nutrition.

Thymidylate synthase (TS) protein expression as a prognostic factor in advanced colorectal cancer: a comparison with TS mRNA expression.

PubMed

Nakagawa, Tateo; Shimada, Mitsuo; Kurita, Nobuhiro; Iwata, Takashi; Nishioka, Masanori; Yoshikawa, Kozo; Higashijima, Jun; Utsunomiya, Tohru

2012-06-01

The role of intratumoral thymidylate synthase (TS) mRNA or protein expression is still controversial and little has been reported regarding relation of them in colorectal cancer. Forty-six patients with advanced colorectal cancer who underwent surgical resection were included. TS mRNA expression was determined by the Danenberg tumor profile method based on laser-captured micro-dissection of the tumor cells. TS protein expression was evaluated using immunohistochemical staining. TS mRNA expression tended to relate TS protein expression. Statistical significance was not found in overall survival between the TS mRNA high group and low group regardless of performing adjuvant chemotherapy. The overall survival in the TS protein negative group was significantly higher than that in positive group in all and the patients without adjuvant chemotherapy. Multivariate analysis showed TS protein expression was as an independent prognostic factor. TS protein expression tends to be related TS mRNA expression and is an independent prognostic factor in advanced colorectal cancer.

Links between Dietary Protein Sources, the Gut Microbiota, and Obesity.

PubMed

Madsen, Lise; Myrmel, Lene S; Fjære, Even; Liaset, Bjørn; Kristiansen, Karsten

2017-01-01

The association between the gut microbiota and obesity is well documented in both humans and in animal models. It is also demonstrated that dietary factors can change the gut microbiota composition and obesity development. However, knowledge of how diet, metabolism and gut microbiota mutually interact and modulate energy metabolism and obesity development is still limited. Epidemiological studies indicate an association between intake of certain dietary protein sources and obesity. Animal studies confirm that different protein sources vary in their ability to either prevent or induce obesity. Different sources of protein such as beans, vegetables, dairy, seafood, and meat differ in amino acid composition. Further, the type and level of other factors, such as fatty acids and persistent organic pollutants (POPs) vary between dietary protein sources. All these factors can modulate the composition of the gut microbiota and may thereby influence their obesogenic properties. This review summarizes evidence of how different protein sources affect energy efficiency, obesity development, and the gut microbiota, linking protein-dependent changes in the gut microbiota with obesity.

Links between Dietary Protein Sources, the Gut Microbiota, and Obesity

PubMed Central

Madsen, Lise; Myrmel, Lene S.; Fjære, Even; Liaset, Bjørn; Kristiansen, Karsten

2017-01-01

The association between the gut microbiota and obesity is well documented in both humans and in animal models. It is also demonstrated that dietary factors can change the gut microbiota composition and obesity development. However, knowledge of how diet, metabolism and gut microbiota mutually interact and modulate energy metabolism and obesity development is still limited. Epidemiological studies indicate an association between intake of certain dietary protein sources and obesity. Animal studies confirm that different protein sources vary in their ability to either prevent or induce obesity. Different sources of protein such as beans, vegetables, dairy, seafood, and meat differ in amino acid composition. Further, the type and level of other factors, such as fatty acids and persistent organic pollutants (POPs) vary between dietary protein sources. All these factors can modulate the composition of the gut microbiota and may thereby influence their obesogenic properties. This review summarizes evidence of how different protein sources affect energy efficiency, obesity development, and the gut microbiota, linking protein-dependent changes in the gut microbiota with obesity. PMID:29311977

Role of activator protein-1 on the effect of arginine-glycine-aspartic acid containing peptides on transforming growth factor-beta1 promoter activity.

PubMed

Ruiz-Torres, M P; Perez-Rivero, G; Diez-Marques, M L; Griera, M; Ortega, R; Rodriguez-Puyol, M; Rodríguez-Puyol, D

2007-01-01

While arginine-glycine-aspartic acid-based peptidomimetics have been employed for the treatment of cardiovascular disorders and cancer, their use in other contexts remains to be explored. Arginine-glycine-aspartic acid-serine induces Transforming growth factor-beta1 transcription in human mesangial cells, but the molecular mechanisms involved have not been studied extensively. We explored whether this effect could be due to Activator protein-1 activation and studied the potential pathways involved. Addition of arginine-glycine-aspartic acid-serine promoted Activator protein-1 binding to its cognate sequence within the Transforming growth factor-beta1 promoter as well as c-jun and c-fos protein abundance. Moreover, this effect was suppressed by curcumin, a c-Jun N terminal kinase inhibitor, and was absent when the Activator protein-1 cis-regulatory element was deleted. Activator protein-1 binding was dependent on the activity of integrin linked kinase, as transfection with a dominant negative mutant suppressed both Activator protein-1 binding and c-jun and c-fos protein increment. Integrin linked kinase was, in turn, dependent on Phosphoinositol-3 kinase activity. Arginine-glycine-aspartic acid-serine stimulated Phosphoinositol-3 kinase activity, and Transforming growth factor-beta1 promoter activation was abrogated by the use of Phosphoinositol-3 kinase specific inhibitors. In summary, we propose that arginine-glycine-aspartic acid-serine activates Integrin linked kinase via the Phosphoinositol-3 kinase pathway and this leads to activation of c-jun and c-fos and increased Activator protein-1 binding and Transforming growth factor-beta1 promoter activity. These data may contribute to understand the molecular mechanisms involved in the cellular actions of arginine-glycine-aspartic acid-related peptides and enhance their relevance as these products evolve into clinical therapeutic use.

Streptococcus agalactiae Non-Pilus, Cell Wall-Anchored Proteins: Involvement in Colonization and Pathogenesis and Potential as Vaccine Candidates

PubMed Central

Pietrocola, Giampiero; Arciola, Carla Renata; Rindi, Simonetta; Montanaro, Lucio; Speziale, Pietro

2018-01-01

Group B Streptococcus (GBS) remains an important etiological agent of several infectious diseases including neonatal septicemia, pneumonia, meningitis, and orthopedic device infections. This pathogenicity is due to a variety of virulence factors expressed by Streptococcus agalactiae. Single virulence factors are not sufficient to provoke a streptococcal infection, which is instead promoted by the coordinated activity of several pathogenicity factors. Such determinants, mostly cell wall-associated and secreted proteins, include adhesins that mediate binding of the pathogen to host extracellular matrix/plasma ligands and cell surfaces, proteins that cooperate in the invasion of and survival within host cells and factors that neutralize phagocytosis and/or modulate the immune response. The genome-based approaches and bioinformatics tools and the extensive use of biophysical and biochemical methods and animal model studies have provided a great wealth of information on the molecular structure and function of these virulence factors. In fact, a number of new GBS surface-exposed or secreted proteins have been identified (GBS immunogenic bacterial adhesion protein, leucine-rich repeat of GBS, serine-rich repeat proteins), the three-dimensional structures of known streptococcal proteins (αC protein, C5a peptidase) have been solved and an understanding of the pathogenetic role of “old” and new determinants has been better defined in recent years. Herein, we provide an update of our current understanding of the major surface cell wall-anchored proteins from GBS, with emphasis on their biochemical and structural properties and the pathogenetic roles they may have in the onset and progression of host infection. We also focus on the antigenic profile of these compounds and discuss them as targets for therapeutic intervention. PMID:29686667

Analysis of In Vivo Chromatin and Protein Interactions of Arabidopsis Transcript Elongation Factors.

PubMed

Pfab, Alexander; Antosz, Wojciech; Holzinger, Philipp; Bruckmann, Astrid; Griesenbeck, Joachim; Grasser, Klaus D

2017-01-01

A central step to elucidate the function of proteins commonly comprises the analysis of their molecular interactions in vivo. For nuclear regulatory proteins this involves determining protein-protein interactions as well as mapping of chromatin binding sites. Here, we present two protocols to identify protein-protein and chromatin interactions of transcript elongation factors (TEFs) in Arabidopsis. The first protocol (Subheading 3.1) describes protein affinity-purification coupled to mass spectrometry (AP-MS) that utilizes suspension cultured cells as experimental system. This approach provides an unbiased view of proteins interacting with epitope-tagged TEFs. The second protocol (Subheading 3.2) depicts details about a chromatin immunoprecipitation (ChIP) procedure to characterize genomic binding sites of TEFs. These methods should be valuable tools for the analysis of a broad variety of nuclear proteins.

The strategy of fusion genes construction determines efficient expression of introduced transcription factors.

PubMed

Adamus, Tomasz; Konieczny, Paweł; Sekuła, Małgorzata; Sułkowski, Maciej; Majka, Marcin

2014-01-01

The main goal in gene therapy and biomedical research is an efficient transcription factors (TFs) delivery system. SNAIL, a zinc finger transcription factor, is strongly involved in tumor, what makes its signaling pathways an interesting research subject. The necessity of tracking activation of intracellular pathways has prompted fluorescent proteins usage as localization markers. Advanced molecular cloning techniques allow to generate fusion proteins from fluorescent markers and transcription factors. Depending on fusion strategy, the protein expression levels and nuclear transport ability are significantly different. The P2A self-cleavage motif through its cleavage ability allows two single proteins to be simultaneously expressed. The aim of this study was to compare two strategies for introducing a pair of genes using expression vector system. We have examined GFP and SNAI1 gene fusions by comprising common nucleotide polylinker (multiple cloning site) or P2A motif in between them, resulting in one fusion or two independent protein expressions respectively. In each case transgene expression levels and translation efficiency as well as nuclear localization of expressed protein have been analyzed. Our data showed that usage of P2A motif provides more effective nuclear transport of SNAIL transcription factor than conventional genes linker. At the same time the fluorescent marker spreads evenly in subcellular space.

Genome-wide analysis of gene expression and protein secretion of Babesia canis during virulent infection identifies potential pathogenicity factors.

PubMed

Eichenberger, Ramon M; Ramakrishnan, Chandra; Russo, Giancarlo; Deplazes, Peter; Hehl, Adrian B

2017-06-13

Infections of dogs with virulent strains of Babesia canis are characterized by rapid onset and high mortality, comparable to complicated human malaria. As in other apicomplexan parasites, most Babesia virulence factors responsible for survival and pathogenicity are secreted to the host cell surface and beyond where they remodel and biochemically modify the infected cell interacting with host proteins in a very specific manner. Here, we investigated factors secreted by B. canis during acute infections in dogs and report on in silico predictions and experimental analysis of the parasite's exportome. As a backdrop, we generated a fully annotated B. canis genome sequence of a virulent Hungarian field isolate (strain BcH-CHIPZ) underpinned by extensive genome-wide RNA-seq analysis. We find evidence for conserved factors in apicomplexan hemoparasites involved in immune-evasion (e.g. VESA-protein family), proteins secreted across the iRBC membrane into the host bloodstream (e.g. SA- and Bc28 protein families), potential moonlighting proteins (e.g. profilin and histones), and uncharacterized antigens present during acute crisis in dogs. The combined data provides a first predicted and partially validated set of potential virulence factors exported during fatal infections, which can be exploited for urgently needed innovative intervention strategies aimed at facilitating diagnosis and management of canine babesiosis.

Functional interaction of the DNA-binding transcription factor Sp1 through its DNA-binding domain with the histone chaperone TAF-I.

PubMed

Suzuki, Toru; Muto, Shinsuke; Miyamoto, Saku; Aizawa, Kenichi; Horikoshi, Masami; Nagai, Ryozo

2003-08-01

Transcription involves molecular interactions between general and regulatory transcription factors with further regulation by protein-protein interactions (e.g. transcriptional cofactors). Here we describe functional interaction between DNA-binding transcription factor and histone chaperone. Affinity purification of factors interacting with the DNA-binding domain of the transcription factor Sp1 showed Sp1 to interact with the histone chaperone TAF-I, both alpha and beta isoforms. This interaction was specific as Sp1 did not interact with another histone chaperone CIA nor did other tested DNA-binding regulatory factors (MyoD, NFkappaB, p53) interact with TAF-I. Interaction of Sp1 and TAF-I occurs both in vitro and in vivo. Interaction with TAF-I results in inhibition of DNA-binding, and also likely as a result of such, inhibition of promoter activation by Sp1. Collectively, we describe interaction between DNA-binding transcription factor and histone chaperone which results in negative regulation of the former. This novel regulatory interaction advances our understanding of the mechanisms of eukaryotic transcription through DNA-binding regulatory transcription factors by protein-protein interactions, and also shows the DNA-binding domain to mediate important regulatory interactions.

Discovery and Development of Kelch-like ECH-Associated Protein 1. Nuclear Factor Erythroid 2-Related Factor 2 (KEAP1:NRF2) Protein-Protein Interaction Inhibitors: Achievements, Challenges, and Future Directions.

PubMed

Jiang, Zheng-Yu; Lu, Meng-Chen; You, Qi-Dong

2016-12-22

The transcription factor Nrf2 is the primary regulator of the cellular defense system, and enhancing Nrf2 activity has potential usages in various diseases, especially chronic age-related and inflammatory diseases. Recently, directly targeting Keap1-Nrf2 protein-protein interaction (PPI) has been an emerging strategy to selectively and effectively activate Nrf2. This Perspective summarizes the progress in the discovery and development of Keap1-Nrf2 PPI inhibitors, including the Keap1-Nrf2 regulatory mechanisms, biochemical techniques for inhibitor identification, and approaches for identifying peptide and small-molecule inhibitors, as well as discusses privileged structures and future directions for further development of Keap1-Nrf2 PPI inhibitors.

Identification of host factors potentially involved in RTM-mediated resistance during potyvirus long distance movement.

PubMed

Sofer, Luc; Cabanillas, Daniel Garcia; Gayral, Mathieu; Téplier, Rachèle; Pouzoulet, Jérôme; Ducousso, Marie; Dufin, Laurène; Bréhélin, Claire; Ziegler-Graff, Véronique; Brault, Véronique; Revers, Frédéric

2017-07-01

The long distance movement of potyviruses is a poorly understood step of the viral cycle. Only factors inhibiting this process, referred to as "Restricted TEV Movement" (RTM), have been identified in Arabidopsis thaliana. On the virus side, the potyvirus coat protein (CP) displays determinants required for long-distance movement and for RTM-based resistance breaking. However, the potyvirus CP was previously shown not to interact with the RTM proteins. We undertook the identification of Arabidopsis factors which directly interact with either the RTM proteins or the CP of lettuce mosaic virus (LMV). An Arabidopsis cDNA library generated from companion cells was screened with LMV CP and RTM proteins using the yeast two-hybrid system. Fourteen interacting proteins were identified. Two of them were shown to interact with CP and the RTM proteins suggesting that a multiprotein complex could be formed between the RTM proteins and virions or viral ribonucleoprotein complexes. Co-localization experiments in Nicotiana benthamiana showed that most of the viral and cellular protein pairs co-localized at the periphery of chloroplasts which suggests a putative role for plastids in this process.

Maize endosperm-specific transcription factors O2 and PBF network the regulation of protein and starch synthesis

PubMed Central

Zhang, Zhiyong; Zheng, Xixi; Yang, Jun; Messing, Joachim; Wu, Yongrui

2016-01-01

The maize endosperm-specific transcription factors opaque2 (O2) and prolamine-box binding factor (PBF) regulate storage protein zein genes. We show that they also control starch synthesis. The starch content in the PbfRNAi and o2 mutants was reduced by ∼5% and 11%, respectively, compared with normal genotypes. In the double-mutant PbfRNAi;o2, starch was decreased by 25%. Transcriptome analysis reveals that >1,000 genes were affected in each of the two mutants and in the double mutant; these genes were mainly enriched in sugar and protein metabolism. Pyruvate orthophosphate dikinase 1 and 2 (PPDKs) and starch synthase III (SSIII) are critical components in the starch biosynthetic enzyme complex. The expression of PPDK1, PPDK2, and SSIII and their protein levels are further reduced in the double mutants as compared with the single mutants. When the promoters of these genes were analyzed, we found a prolamine box and an O2 box that can be additively transactivated by PBF and O2. Starch synthase IIa (SSIIa, encoding another starch synthase for amylopectin) and starch branching enzyme 1 (SBEI, encoding one of the two main starch branching enzymes) are not directly regulated by PBF and O2, but their protein levels are significantly decreased in the o2 mutant and are further decreased in the double mutant, indicating that o2 and PbfRNAi may affect the levels of some other transcription factor(s) or mRNA regulatory factor(s) that in turn would affect the transcript and protein levels of SSIIa and SBEI. These findings show that three important traits—nutritional quality, calories, and yield—are linked through the same transcription factors. PMID:27621432

Rapid Modulation of Protein Expression in the Rat Hippocampus Following Deep Brain Stimulation of the Fornix.

PubMed

Gondard, Elise; Chau, Hien N; Mann, Amandeep; Tierney, Travis S; Hamani, Clement; Kalia, Suneil K; Lozano, Andres M

2015-01-01

The forniceal area is currently being evaluated as a target for deep brain stimulation (DBS) to improve cognitive function in patients with Alzheimer's disease. The molecular changes at downstream targets within the stimulated circuit are unknown. To analyze the modulation of hippocampal protein expression following 1 h of fornix DBS in the rat. Animals underwent bilateral forniceal DBS for 1 h and sacrificed at different time-points after the initiation of the stimulation (1 h, 2.5 h, 5 h, 25 h). Bilateral hippocampi were isolated for western blot analyses. Forniceal DBS led to a dramatic elevation of cFos post-stimulation, suggesting that forniceal DBS activates the hippocampus. There was also a significant increase in candidate proteins including several trophic factors, such as brain derived neurotrophic factor (BDNF) and vascular endothelial growth factor (VEGF) but not glial cell-derived neurotrophic factor (GDNF). There was in addition, increased expression of the synaptic markers growth associated protein 43 (GAP-43), synaptophysin and α-synuclein. No changes were observed at the studied time-points in Alzheimer's-related proteins including amyloid precursor protein (APP), tau, phosphorylated tau (ptau), or selected chaperone proteins (HSP40, HSP70 and CHIP). Forniceal DBS triggers hippocampal activity and rapidly modulate the expression of neurotrophic factors and markers of synaptic plasticity known to play key roles in memory processing. The clinical effects of DBS of the fornix may, in part, be mediated by producing changes in the expression of these proteins. Copyright © 2015 Elsevier Inc. All rights reserved.

Some growth factors stimulate cultured adult rabbit ventricular myocyte hypertrophy in the absence of mechanical loading

NASA Technical Reports Server (NTRS)

Decker, R. S.; Cook, M. G.; Behnke-Barclay, M.; Decker, M. L.

1995-01-01

Cultured adult rabbit cardiac myocytes treated with recombinant growth factors display enhanced rates of protein accumulation (ie, growth) in response to insulin and insulin-like growth factors (IGFs), but epidermal growth factor, acidic or basic fibroblast growth factor, and platelet-derived growth factor failed to increase contractile protein synthesis or growth of the heart cells. Insulin and IGF-1 increased growth rates by stimulating anabolic while simultaneously inhibiting catabolic pathways, whereas IGF-2 elevated growth modestly by apparently inhibiting lysosomal proteolysis. Neutralizing antibodies directed against either IGF-1 or IGF-2 or IGF binding protein 3 blocked protein accumulation. A monoclonal antibody directed against the IGF-1 receptor also inhibited changes in protein turnover provoked by recombinant human IGF-1 but not IGF-2. Of the other growth factors tested, only transforming growth factor-beta 1 increased the fractional rate of myosin heavy chain (MHC) synthesis, with beta-MHC synthesis being elevated and alpha-MHC synthesis being suppressed. However, the other growth factors were able to modestly stimulate the rate of DNA synthesis in this preparation. Bromodeoxyuridine labeling revealed that these growth factors increased DNA synthesis in myocytes and nonmyocytes alike, but the heart cells displayed neither karyokinesis or cytokinesis. In contrast, cocultures of cardiac myocytes and nonmyocytes and nonmyocyte-conditioned culture medium failed to enhance the rate of cardiac MHC synthesis or its accumulation, implying that quiescent heart cells do not respond to "conditioning" by cardiac nonmyocytes. These findings demonstrated that insulin and the IGFs promote passively loaded cultured adult rabbit heart cells to hypertrophy but suggest that other growth factors tested may be limited in this regard.

Gradient elution behavior of proteins in hydrophobic interaction chromatography with U-shaped retention factor curves.

PubMed

Creasy, Arch; Lomino, Joseph; Barker, Gregory; Khetan, Anurag; Carta, Giorgio

2018-04-27

Protein retention in hydrophobic interaction chromatography is described by the solvophobic theory as a function of the kosmostropic salt concentration. In general, an increase in salt concentration drives protein partitioning to the hydrophobic surface while a decrease reduces it. In some cases, however, protein retention also increases at low salt concentrations resulting in a U-shaped retention factor curve. During gradient elution the salt concentration is gradually decreased from a high value thereby reducing the retention factor and increasing the protein chromatographic velocity. For these conditions, a steep gradient can overtake the protein in the column, causing it to rebind. Two dynamic models, one based on the local equilibrium theory and the other based on the linear driving force approximation, are presented. We show that the normalized gradient slope determines whether the protein elutes in the gradient, partially elutes, or is trapped in the column. Experimental results are presented for two different monoclonal antibodies and for lysozyme on Capto Phenyl (High Sub) resin. One of the mAbs and lysozyme exhibit U-shaped retention factor curves and for each, we determine the critical gradient slope beyond which 100% recovery is no longer possible. Elution with a reverse gradient is also demonstrated at low salt concentrations for these proteins. Understanding this behavior has implications in the design of gradient elution since the gradient slope impacts protein recovery. Copyright © 2018 Elsevier B.V. All rights reserved.

Intact stable isotope labeled plasma proteins from the SILAC-labeled HepG2 secretome.

PubMed

Mangrum, John B; Martin, Erika J; Brophy, Donald F; Hawkridge, Adam M

2015-09-01

The plasma proteome remains an attractive biospecimen for MS-based biomarker discovery studies. The success of these efforts relies on the continued development of quantitative MS-based proteomics approaches. Herein we report the use of the SILAC-labeled HepG2 secretome as a source for stable isotope labeled plasma proteins for quantitative LC-MS/MS measurements. The HepG2 liver cancer cell line secretes the major plasma proteins including serum albumin, apolipoproteins, protease inhibitors, coagulation factors, and transporters that represent some of the most abundant proteins in plasma. The SILAC-labeled HepG2 secretome was collected, spiked into human plasma (1:1 total protein), and then processed for LC-MS/MS analysis. A total of 62 and 56 plasma proteins were quantified (heavy:light (H/L) peptide pairs) from undepleted and depleted (serum albumin and IgG), respectively, with log2 H/L = ± 6. Major plasma proteins quantified included albumin, apolipoproteins (e.g., APOA1, APOA2, APOA4, APOB, APOC3, APOE, APOH, and APOM), protease inhibitors (e.g., A2M and SERPINs), coagulation factors (e.g., Factor V, Factor X, fibrinogen), and transport proteins (e.g., TTR). The average log2 H/L values for shared plasma proteins in both undepleted and depleted plasma samples were 0.43 and 0.44, respectively. This work further expands the SILAC strategy into MS-based biomarker discovery of clinical biospecimens. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Active G protein-coupled receptors (GPCR), matrix metalloproteinases 2/9 (MMP2/9), heparin-binding epidermal growth factor (hbEGF), epidermal growth factor receptor (EGFR), erbB2, and insulin-like growth factor 1 receptor (IGF-1R) are necessary for trenbolone acetate-induced alterations in protein turnover rate of fused bovine satellite cell cultures.

PubMed

Thornton, K J; Kamanga-Sollo, E; White, M E; Dayton, W R

2016-06-01

Trenbolone acetate (TBA), a testosterone analog, increases protein synthesis and decreases protein degradation in fused bovine satellite cell (BSC) cultures. However, the mechanism through which TBA alters these processes remains unknown. Recent studies indicate that androgens improve rate and extent of muscle growth through a nongenomic mechanism involving G protein-coupled receptors (GPCR), matrix metalloproteinases (MMP), heparin-binding epidermal growth factor (hbEGF), the epidermal growth factor receptor (EGFR), erbB2, and the insulin-like growth factor-1 receptor (IGF-1R). We hypothesized that TBA activates GPCR, resulting in activation of MMP2/9 that releases hbEGF, which activates the EGFR and/or erbB2. To determine whether the proposed nongenomic pathway is involved in TBA-mediated alterations in protein turnover, fused BSC cultures were treated with TBA in the presence or absence of inhibitors for GPCR, MMP2/9, hbEGF, EGFR, erbB2, or IGF-1R, and resultant protein synthesis and degradation rates were analyzed. Assays were replicated at least 9 times for each inhibitor experiment utilizing BSC cultures obtained from at least 3 different steers that had no previous exposure to steroid compounds. As expected, fused BSC cultures treated with 10 n TBA exhibited increased ( < 0.05) protein synthesis rates and decreased ( < 0.05) protein degradation rates when compared to control cultures. Treatment of fused BSC cultures with 10 n TBA in the presence of inhibitors for GPCR, MMP2/9, hbEGF, EGFR, erbB2, or IGF-1R suppressed ( < 0.05) TBA-mediated increases in protein synthesis rate. Alternatively, inhibition of GPCR, MMP2/9, hbEGF, EGFR, erbB2, or IGF-1R in the presence of 10 n TBA each had no ( > 0.05) effect on TBA-mediated decreases in protein degradation. However, inhibition of both EGFR and erbB2 in the presence of 10 n TBA resulted in decreased ( < 0.05) ability of TBA to decrease protein degradation rate. Additionally, fused BSC cultures treated with 10 n TBA exhibit increased ( < 0.05) pAKT protein levels. These data indicate the TBA-mediated increases in protein synthesis likely involve GPCR, MMP2/9, hbEGF, EGFR, erbB2, and IGF-1R. However, the mechanism through which TBA mediates changes in protein degradation is different and appears to involve only the EGFR and erbB2. Furthermore, it appears the protein kinase B pathway is involved in TBA's effects on fused BSC cultures.

Immunogenicity and immune tolerance coagulation Factors VIII and IX.

PubMed

Rup, B

2003-01-01

Some of the major issues related to the development and control of antibodies that occur during treatment of haemophilia with replacement factors (Factor VIII and Factor IX) are reviewed. Information on analytical issues, immunogenicity, and immune tolerance may be applicable to the study of other therapeutic proteins. Conversely, new information obtained from evaluation of other therapeutic protein products may address issues that remain unresolved for Factor VIII and FIX replacement therapy.

Regulatory role of the 90-kDa-heat-shock protein (Hsp90) and associated factors on gene expression.

PubMed

Erlejman, Alejandra G; Lagadari, Mariana; Toneatto, Judith; Piwien-Pilipuk, Graciela; Galigniana, Mario D

2014-02-01

The term molecular chaperone was first used to describe the ability of nucleoplasmin to prevent the aggregation of histones with DNA during the assembly of nucleosomes. Subsequently, the name was extended to proteins that mediate the post-translational assembly of oligomeric complexes protecting them from denaturation and/or aggregation. Hsp90 is a 90-kDa molecular chaperone that represents the major soluble protein of the cell. In contrast to most conventional chaperones, Hsp90 functions as a refined sensor of protein function and its principal role in the cell is to facilitate biological activity to properly folded client proteins that already have a preserved tertiary structure. Consequently, Hsp90 is related to basic cell functions such as cytoplasmic transport of soluble proteins, translocation of client proteins to organelles, and regulation of the biological activity of key signaling factors such as protein kinases, ubiquitin ligases, steroid receptors, cell cycle regulators, and transcription factors. A growing amount of evidence links the protective action of this molecular chaperone to mechanisms related to posttranslational modifications of soluble nuclear factors as well as histones. In this article, we discuss some aspects of the regulatory action of Hsp90 on transcriptional regulation and how this effect could have impacted genetic assimilation mechanism in some organisms. Copyright © 2013 Elsevier B.V. All rights reserved.

Nutrition and physical activity for the prevention and treatment of age-related sarcopenia.

PubMed

Bosaeus, Ingvar; Rothenberg, Elisabet

2016-05-01

Sarcopenia, defined as loss of skeletal muscle mass and function, is associated with adverse outcomes such as physical disability, impaired quality of life and increased mortality. Several mechanisms are involved in the development of sarcopenia. Potentially modifiable factors include nutrition and physical activity. Protein metabolism is central to the nutritional issues, along with other potentially modifying nutritional factors as energy balance and vitamin D status. An increasing but still incomplete knowledge base has generated recent recommendations on an increased protein intake in the elderly. Several factors beyond the total amount of protein consumed emerge as potentially important in this context. A recent summit examined three hypotheses: (1) A meal threshold; habitually consuming 25-30 g protein at breakfast, lunch and dinner provides sufficient protein to effectively stimulate muscle protein anabolism; (2) Protein quality; including high-quality protein at each meal improves postprandial muscle protein synthesis; and (3) performing physical activity in close temporal proximity to a high-quality protein meal enhances muscle anabolism. Optimising the potential for muscle protein anabolism by consuming an adequate amount of high-quality protein at each meal, in combination with physical activity, appears as a promising strategy to prevent or delay the onset of sarcopenia. However, results of interventions are inconsistent, and well-designed, standardised studies evaluating exercise or nutrition interventions are needed before guidelines can be developed for the prevention and treatment of age-related sarcopenia.

Secreted Proteins Defy the Expression Level–Evolutionary Rate Anticorrelation

PubMed Central

Feyertag, Felix; Berninsone, Patricia M.; Alvarez-Ponce, David

2017-01-01

The rates of evolution of the proteins of any organism vary across orders of magnitude. A primary factor influencing rates of protein evolution is expression. A strong negative correlation between expression levels and evolutionary rates (the so-called E–R anticorrelation) has been observed in virtually all studied organisms. This effect is currently attributed to the abundance-dependent fitness costs of misfolding and unspecific protein–protein interactions, among other factors. Secreted proteins are folded in the endoplasmic reticulum, a compartment where chaperones, folding catalysts, and stringent quality control mechanisms promote their correct folding and may reduce the fitness costs of misfolding. In addition, confinement of secreted proteins to the extracellular space may reduce misinteractions and their deleterious effects. We hypothesize that each of these factors (the secretory pathway quality control and extracellular location) may reduce the strength of the E–R anticorrelation. Indeed, here we show that among human proteins that are secreted to the extracellular space, rates of evolution do not correlate with protein abundances. This trend is robust to controlling for several potentially confounding factors and is also observed when analyzing protein abundance data for 6 human tissues. In addition, analysis of mRNA abundance data for 32 human tissues shows that the E–R correlation is always less negative, and sometimes nonsignificant, in secreted proteins. Similar observations were made in Caenorhabditis elegans and in Escherichia coli, and to a lesser extent in Drosophila melanogaster, Saccharomyces cerevisiae and Arabidopsis thaliana. Our observations contribute to understand the causes of the E–R anticorrelation. PMID:28007979

Endogenous versus Exogenous Growth Factor Regulation of Articular Chondrocytes

PubMed Central

Shi, Shuiliang; Chan, Albert G.; Mercer, Scott; Eckert, George J.; Trippel, Stephen B.

2014-01-01

Anabolic growth factors that regulate the function of articular chondrocytes are candidates for articular cartilage repair. Such factors may be delivered by pharmacotherapy in the form of exogenous proteins, or by gene therapy as endogenous proteins. It is unknown whether delivery method influences growth factor effectiveness in regulating articular chondrocyte reparative functions. We treated adult bovine articular chondrocytes with exogenous recombinant insulin-like growth factor-I (IGF-I) and transforming growth factor-beta1 (TGF-β1), or with the genes encoding these growth factors for endogenous production. Treatment effects were measured as change in chondrocyte DNA content, glycosaminoglycan production, and aggrecan gene expression. We found that IGF-I stimulated chondrocyte biosynthesis similarly when delivered by either exogenous or endogenous means. In contrast, exogenous TGF-ß1 stimulated these reparative functions, while endogenous TGF-ß1 had little effect. Endogenous TGF-ß1 became more bioactive following activation of the transgene protein product. These data indicate that effective mechanisms of growth factor delivery for articular cartilage repair may differ for different growth factors. In the case of IGF-I, gene therapy or protein therapy appear to be viable options. In contrast, TGF-ß1 gene therapy may be constrained by a limited ability of chondrocytes to convert latent complexes to an active form. PMID:24105960