[The relationship between lactate clearance rate and delayed encephalopathy after acute carbon monoxide poisoning].

PubMed

Wang, Wei-zhan; Ma, Guo-ying; Zhao, Ling-jun; Li, Jing; Wang, Pu; Li, Ya-qin; Xiao, Qing-mian; Liu, Yong-jian

2013-10-01

To study the relationship between lactate clearance rate (LCR) and prognosis after acute carbon monoxide poisoning in patients with delayed encephalopathy (DEACMP). Data from 354 patients with acute severe carbon monoxide poisoning (ASCOP) were retrospectively analyzed. The patients were divided into hyperlactacidemia group (arterial lactic acid > 2 mmol/L, n=263) and low lactic acidosis group (arterial lactate ≤2 mmol/L, n=91) according to the blood lactic acid level at admission. Arterial blood (1 mL) was collected from all patients before and 6, 24, 72 hours after treatment at ambient air, and arterial blood lactic acid was determined, and LCR was calculated. The initial level of blood lactic acid and LCR at 6, 24, 72 hours were compared between two groups. At the same time, the patients with hyperlactacidemia were divided into high LCR group (LCR more than 10%, n=101) and low LCR group (LCR less than or equal to 10%, n=162) according to 6-hour LCR, and the incidence of DEACMP was compared between two groups. The relationship between LCR and the incidence of DEACMP was analyzed with Spearman linear correlation analysis. The risk factors associated with DEACMP were analyzed with logistic regression analysis. The initial level of blood lactic acid (2.73±0.57 mmol/L vs. 1.69±0.20 mmol/L, t=5.327, P=0.001) and LCR at 6, 24, 72 hours [6 hours: (9.0±2.4)% vs. (1.2±0.6)%, t=9.468, P=0.001; 24 hours: (8.6±3.7)% vs. (1.2±0.4)%, t=4.889, P=0.001; 72 hours: (14.0±3.9)% vs. (1.7±1.0)%, t=5.211, P=0.001] in hyperlactacidemia group were significantly higher than those in low lactic acidosis group. The initial level of blood lactic acid in high LCR group was significantly lower than that in low LCR group (2.41±0.23 mmol/L vs. 2.92±0.63 mmol/L, t=2.429, P=0.023), and LCR at 6 hours and 24 hours were significantly higher than those in low LCR group [6 hours: (11.0±1.2)% vs. (8.0±2.1)%, t=4.487, P=0.001; 24 hours: (12.2±3.0)% vs. (6.3±1.8)%, t=6.264, P=0.001]. But there was no difference in 72-hour LCR between high LCR group and low LCR group [(14.1±3.6)% vs. (13.9±4.1)%, t=0.182, P=0.857]. The incidence of DEACMP in high LCR group was significantly lower than that in low LCR group [15.8% (16/101) vs. 61.1% (99/162), χ(2)=51.814, P=0.001]. The blood LCR at early period (6, 24, 72 hours) in ASCOP patients with hyperlactacidemia was negatively correlated with the incidence of DEACMP (r1=-0.493, P1=0.011; r2=-0.408, P2=0.038; r3=-0.428, P3=0.029). Logistic regression analysis showed that LRC at 6 hours and 24 hours [odds ratio (OR) was 2.701, 1.070, P value was 0.035, 0.001], long-time coma (OR=1.537, P=0.068), contact carbon monoxide (CO) long time (OR=2.686, P=0.014), age (OR=1.464, P=0.017), acute carbon monoxide complications (OR=1.363, P=0.072) patients with ASCOP had an increased risk of DEACMP. LCR is helpful for the assess of DEACMP patients severity, for the treatment guide and for prognosis judgement.

Yersinia Type III Secretion System Master Regulator LcrF

PubMed Central

Schwiesow, Leah; Lam, Hanh

2015-01-01

Many Gram-negative pathogens express a type III secretion (T3SS) system to enable growth and survival within a host. The three human-pathogenic Yersinia species, Y. pestis, Y. pseudotuberculosis, and Y. enterocolitica, encode the Ysc T3SS, whose expression is controlled by an AraC-like master regulator called LcrF. In this review, we discuss LcrF structure and function as well as the environmental cues and pathways known to regulate LcrF expression. Similarities and differences in binding motifs and modes of action between LcrF and the Pseudomonas aeruginosa homolog ExsA are summarized. In addition, we present a new bioinformatics analysis that identifies putative LcrF binding sites within Yersinia target gene promoters. PMID:26644429

Serotonin in the solitary tract nucleus shortens the laryngeal chemoreflex in anesthetized neonatal rats

PubMed Central

Donnelly, William T.; Bartlett, Donald; Leiter, J.C.

2017-01-01

The laryngeal chemoreflex (LCR), an airway protective reflex that causes apnea and bradycardia, has long been suspected as an initiating event in the sudden infant death syndrome (SIDS). Serotonin (5-HT) and 5-HT receptors may be deficient in the brainstems of babies who die of SIDS, and 5-HT seems to be important in terminating apneas directly or in causing arousals or as part of the process of autoresuscitation. We hypothesized that 5-HT in the brainstem would limit the duration of the LCR. We studied anesthetized rat pups between 7 and 21 days of age and made microinjections into the cisterna magna or into the nucleus of the solitary tract (NTS). Focal, bilateral microinjections of 5-HT into the caudal NTS significantly shortened the LCR. The 5-HT 1a receptor antagonist, WAY 100635, did not affect the LCR consistently, nor did a 5-HT2 receptor antagonist, ketanserin, alter the duration of the LCR. The 5-HT3 specific agonist, 1-(3-chlorophenyl)-biguanide, microinjected bilaterally into the caudal NTS significantly shortened the LCR. Thus, endogenous 5-HT released within the NTS may curtail the respiratory depression that is part of the LCR, and serotonergic shortening of the LCR may be attributed to activation of 5-HT3 receptors within the NTS. 5-HT3 receptors are expressed presynaptically on C-fiber afferents of the superior laryngeal nerve, and serotonergic shortening of the LCR may be mediated presynaptically by enhanced activation of inhibitory interneurons within the NTS that terminate during the LCR. PMID:27121960

Aerobic capacity and hepatic mitochondrial lipid oxidation alters susceptibility for chronic high-fat diet-induced hepatic steatosis.

PubMed

Morris, E Matthew; Meers, Grace M E; Koch, Lauren G; Britton, Steven L; Fletcher, Justin A; Fu, Xiaorong; Shankar, Kartik; Burgess, Shawn C; Ibdah, Jamal A; Rector, R Scott; Thyfault, John P

2016-10-01

Rats selectively bred for high capacity running (HCR) or low capacity running (LCR) display divergence for intrinsic aerobic capacity and hepatic mitochondrial oxidative capacity, both factors associated with susceptibility for nonalcoholic fatty liver disease. Here, we tested if HCR and LCR rats display differences in susceptibility for hepatic steatosis after 16 wk of high-fat diets (HFD) with either 45% or 60% of kcals from fat. HCR rats were protected against HFD-induced hepatic steatosis, whereas only the 60% HFD induced steatosis in LCR rats, as marked by a doubling of liver triglycerides. Hepatic complete fatty acid oxidation (FAO) and mitochondrial respiratory capacity were all lower in LCR compared with HCR rats. LCR rats also displayed lower hepatic complete and incomplete FAO in the presence of etomoxir, suggesting a reduced role for noncarnitine palmitoyltransferase-1-mediated lipid catabolism in LCR versus HCR rats. Hepatic complete FAO and mitochondrial respiration were largely unaffected by either chronic HFD; however, 60% HFD feeding markedly reduced 2-pyruvate oxidation, a marker of tricarboxylic acid (TCA) cycle flux, and mitochondrial complete FAO only in LCR rats. LCR rats displayed lower levels of hepatic long-chain acylcarnitines than HCR rats but maintained similar levels of hepatic acetyl-carnitine levels, further supporting lower rates of β-oxidation, and TCA cycle flux in LCR than HCR rats. Finally, only LCR rats displayed early reductions in TCA cycle genes after the acute initiation of a HFD. In conclusion, intrinsically high aerobic capacity confers protection against HFD-induced hepatic steatosis through elevated hepatic mitochondrial oxidative capacity.

High intrinsic aerobic capacity and pomegranate juice are protective against macrophage atherogenecity: studies in high- vs. low-capacity runner (HCR vs. LCR) rats.

PubMed

Rosenblat, Mira; Volkova, Nina; Abassi, Zaid; Britton, Steven L; Koch, Lauren G; Aviram, Michael

2015-10-01

We studied the rat model system of high- vs. low-capacity runner (HCR vs. LCR) rats to question the atherogenic properties (oxidative stress, triglycerides and cholesterol metabolism) in the rat macrophages, serum, liver and heart. Half of the LCR or HCR rats consumed pomegranate juice (PJ; 15 μmol of gallic acid equivalents/rat/day) for 3 weeks and were compared to placebo-treated rats. At the end of the study blood samples, peritoneal macrophages (RPM), livers, and hearts were harvested from the rats. RPM harvested from HCR vs. LCR demonstrated reduced cellular oxidation (21%), increased paraoxonase 2 activity (28%) and decreased triglycerides mass (44%). Macrophage uptake rates of fluorescein-isothiocyanate-labeled low-density lipoprotein (LDL) or oxidized LDL were significantly lower, by 37% or by 18%, respectively, in HCR vs. LCR RPM. PJ consumption significantly decreased all the above atherogenic parameters with more substantial beneficial effects observed in the LCR vs. the HCR rats (~80% vs. ~40% improvement, respectively). Similar hypo-triglyceridemic pattern was noted in serum from HCR vs. LCR. In contrast to the above results, liver oxidation and triglycerides mass were both minimally increased in HCR vs. LCR rats by 31% and 28%, respectively. In the heart, lipid content was very low, and interestingly, an absence of any significant oxidative stress, along with modest triglyceride accumulation, was observed. We conclude that HCR vs. LCR rats demonstrate reduced atherogenicity, mostly in their macrophages. PJ exerts a further improvement, mostly in macrophages from LCR rats. Copyright © 2015 Elsevier Inc. All rights reserved.

Aerobic capacity and hepatic mitochondrial lipid oxidation alters susceptibility for chronic high-fat diet-induced hepatic steatosis

PubMed Central

Morris, E. Matthew; Meers, Grace M. E.; Koch, Lauren G.; Britton, Steven L.; Fletcher, Justin A.; Fu, Xiaorong; Shankar, Kartik; Burgess, Shawn C.; Ibdah, Jamal A.; Rector, R. Scott

2016-01-01

Rats selectively bred for high capacity running (HCR) or low capacity running (LCR) display divergence for intrinsic aerobic capacity and hepatic mitochondrial oxidative capacity, both factors associated with susceptibility for nonalcoholic fatty liver disease. Here, we tested if HCR and LCR rats display differences in susceptibility for hepatic steatosis after 16 wk of high-fat diets (HFD) with either 45% or 60% of kcals from fat. HCR rats were protected against HFD-induced hepatic steatosis, whereas only the 60% HFD induced steatosis in LCR rats, as marked by a doubling of liver triglycerides. Hepatic complete fatty acid oxidation (FAO) and mitochondrial respiratory capacity were all lower in LCR compared with HCR rats. LCR rats also displayed lower hepatic complete and incomplete FAO in the presence of etomoxir, suggesting a reduced role for noncarnitine palmitoyltransferase-1-mediated lipid catabolism in LCR versus HCR rats. Hepatic complete FAO and mitochondrial respiration were largely unaffected by either chronic HFD; however, 60% HFD feeding markedly reduced 2-pyruvate oxidation, a marker of tricarboxylic acid (TCA) cycle flux, and mitochondrial complete FAO only in LCR rats. LCR rats displayed lower levels of hepatic long-chain acylcarnitines than HCR rats but maintained similar levels of hepatic acetyl-carnitine levels, further supporting lower rates of β-oxidation, and TCA cycle flux in LCR than HCR rats. Finally, only LCR rats displayed early reductions in TCA cycle genes after the acute initiation of a HFD. In conclusion, intrinsically high aerobic capacity confers protection against HFD-induced hepatic steatosis through elevated hepatic mitochondrial oxidative capacity. PMID:27600823

Immunization with recombinant V10 protects cynomolgus macaques from lethal pneumonic plague.

PubMed

Cornelius, Claire A; Quenee, Lauriane E; Overheim, Katie A; Koster, Frederick; Brasel, Trevor L; Elli, Derek; Ciletti, Nancy A; Schneewind, Olaf

2008-12-01

Vaccine and therapeutic strategies that prevent infections with Yersinia pestis have been sought for over a century. Immunization with live attenuated (nonpigmented) strains and immunization with subunit vaccines containing recombinant low-calcium-response V antigen (rLcrV) and recombinant F1 (rF1) antigens are considered effective in animal models. Current antiplague subunit vaccines in development for utilization in humans contain both antigens, either as equal concentrations of the two components (rF1 plus rLcrV) or as a fusion protein (rF1-rLcrV). Here, we show that immunization with either purified rLcrV (a protein at the tip of type III needles) or a variant of this protein, recombinant V10 (rV10) (lacking amino acid residues 271 to 300), alone or in combination with rF1, prevented pneumonic lesions and disease pathogenesis. In addition, passive immunization studies showed that specific antibodies of macaques immunized with rLcrV, rV10, or rF1, either alone or in combination, conferred protection against bubonic plague challenge in mice. Finally, we found that when we compared the reactivities of anti-rLcrV and anti-rV10 immune sera from cynomolgus macaques, BALB/c mice, and brown Norway rats with LcrV-derived peptides, rV10, but not rLcrV immune sera, lacked antibodies recognizing linear LcrV oligopeptides.

Complete TCRα gene locus control region activity in T cells derived in vitro from embryonic stem cells

PubMed Central

Lahiji, Armin; Kučerová-Levisohn, Martina; Lovett, Jordana; Holmes, Roxanne; Zúñiga-Pflücker, Juan Carlos; Ortiz, Benjamin D.

2013-01-01

Locus Control Regions (LCR) are cis-acting gene regulatory elements with the unique, integration site-independent ability to transfer the characteristics of their locus-of-origin’s gene expression pattern to a linked transgene in mice. LCR activities have been discovered in numerous T cell lineage expressed gene loci. These elements can be adapted to the design of stem cell gene therapy vectors that direct robust therapeutic gene expression to the T cell progeny of engineered stem cells. Currently, transgenic mice provide the only experimental approach that wholly supports all the critical aspects of LCR activity. Herein we report manifestation of all key features of mouse T cell receptor (TCR)-α gene LCR function in T cells derived in vitro from mouse embryonic stem cells (ESC). High level, copy number-related TCRα LCR-linked reporter gene expression levels are cell type-restricted in this system, and upregulated during the expected stage transition of T cell development. We further report that de novo introduction of TCRα LCR linked transgenes into existing T cell lines yields incomplete LCR activity. Together, these data indicate that establishing full TCRα LCR activity requires critical molecular events occurring prior to final T-lineage determination. This study additionally validates a novel, tractable and more rapid approach for the study of LCR activity in T cells, and its translation to therapeutic genetic engineering. PMID:23720809

Correlation between voluntary cough and laryngeal cough reflex flows in patients with traumatic brain injury.

PubMed

Lee, Sang Chul; Kang, Seong-Woong; Kim, Min Tae; Kim, Yong Kyun; Chang, Won Hyuk; Im, Sang Hee

2013-08-01

To correlate voluntary cough and laryngeal cough reflex (LCR) flows in patients with traumatic brain injury (TBI). Cross-sectional study. University rehabilitation hospital. Patients with TBI (n=25) and healthy controls (n=48). Not applicable. Peak cough flows (PCFs) and LCR flows were measured using a peak flow meter at the oral-nasal interface. The largest value of 3 attempts was recorded for PCF and LCR, respectively. LCR was elicited by 20% solution of pharmaceutic-grade citric acid dissolved in sterile .15M NaCl solution that was inhaled from a nebulizer. PCF was 447.4 ± 99.0 L/min in the control group and 211.7 ± 58.2 L/min in the patient group. LCR was 209.2 ± 63.8L/min in the control group and 170.0 ± 59.7 L/min in the patient group. Both PCF (P=.000) and LCR (P=.013) were significantly reduced in patients with TBI compared to that of the control group. LCR was strongly related to the PCF in both control (R=.645; P=.000) and patient (R=.711; P=.000) groups. As LCR can be measured as a numerical value and significantly correlates with PCF, LCR can be used to estimate cough ability of patients with TBI who cannot cooperate with PCF measurement. Copyright © 2013 American Congress of Rehabilitation Medicine. Published by Elsevier Inc. All rights reserved.

Amino acid residues 196-225 of LcrV represent a plague protective epitope.

PubMed

Quenee, Lauriane E; Berube, Bryan J; Segal, Joshua; Elli, Derek; Ciletti, Nancy A; Anderson, Deborah; Schneewind, Olaf

2010-02-17

LcrV, a protein that resides at the tip of the type III secretion needles of Yersinia pestis, is the single most important plague protective antigen. Earlier work reported monoclonal antibody MAb 7.3, which binds a conformational epitope of LcrV and protects experimental animals against lethal plague challenge. By screening monoclonal antibodies directed against LcrV for their ability to protect immunized mice against bubonic plague challenge, we examined here the possibility of additional protective epitopes. MAb BA5 protected animals against plague, neutralized the Y. pestis type III secretion pathway and promoted opsonophagocytic clearance of bacteria in blood. LcrV residues 196-225 were necessary and sufficient for MAb BA5 binding. Compared to full-length LcrV, a variant lacking its residues 196-225 retained the ability of eliciting plague protection. These results identify LcrV residues 196-225 as a linear epitope that is recognized by the murine immune system to confer plague protection. Copyright (c) 2009 Elsevier Ltd. All rights reserved.

Amino acid residues 196–225 of LcrV represent a plague protective epitope

PubMed Central

Quenee, Lauriane E.; Berube, Bryan J.; Segal, Joshua; Elli, Derek; Ciletti, Nancy A.; Anderson, Deborah; Schneewind, Olaf

2010-01-01

LcrV, a protein that resides at the tip of the type III secretion needles of Yersinia pestis, is the single most important plague protective antigen. Earlier work reported monoclonal antibody MAb 7.3, which binds a conformational epitope of LcrV and protects experimental animals against lethal plague challenge. By screening monoclonal antibodies directed against LcrV for their ability to protect immunized mice against bubonic plague challenge, we examined here the possibility of additional protective epitopes. MAb BA5 protected animals against plague, neutralized the Y. pestis type III secretion pathway and promoted opsonophagocytic clearance of bacteria in blood. LcrV residues 196–225 were necessary and sufficient for MAb-BA5 binding. Compared to full length LcrV, a variant lacking its residues 196–225 retained the ability of eliciting plague protection. These results identify LcrV residues 196–225 as a linear epitope that is recognized by the murine immune system to confer plague protection. PMID:20005318

Temperature sensing in Yersinia pestis: regulation of yopE transcription by lcrF.

PubMed Central

Hoe, N P; Minion, F C; Goguen, J D

1992-01-01

In Escherichia coli, a yopE::lacZ fusion was found to be regulated by temperature in the presence of the cloned BamHI G fragment of Yersinia pestis plasmid pCD1, which contains the lcrF locus. Increasing the copy number of lcrF relative to that of the yopE reporter had a negligible effect on the induction ratio (26 versus 37 degrees C) but caused large reductions in the absolute levels of yopE transcription. We localized the lcrF gene by monitoring the induction phenotype of BamHI G deletion derivatives. Sequencing revealed an open reading frame capable of encoding a protein of 30.8 kDa. A protein product of this size was detected in a T7 expression system, and LcrF-dependent yopE-specific DNA binding activity was observed. As expected, LcrF exhibited 98% homology to VirF of Yersinia enterocolitica and significant homology to the carboxy termini of other members of the AraC family of transcriptional regulatory proteins. These proteins could be divided into two classes according to function: those regulating operons involved in catabolism of carbon and energy sources and those involved in regulating virulence genes. lcrF::lacZ transcriptional fusions were constructed and analyzed in Y. pestis and E. coli. The activity of the fusions was not affected by the native pCD1 virulence plasmid, an intact lcrF gene, or temperature. Thus, induction of lcrF transcription is not essential for temperature-dependent activation of yopE transcription. A portion of LcrF was found associated with the membrane fraction in E. coli; however, pulse-chase experiments indicated that this result is an artifact of fractionation. Images PMID:1624422

Voluntary wheel running improves adipose tissue immunometabolism in ovariectomized low-fit rats.

PubMed

Zidon, Terese M; Park, Young-Min; Welly, Rebecca J; Woodford, Makenzie L; Scroggins, Rebecca J; Britton, Steven L; Koch, Lauren G; Booth, Frank W; Padilla, Jaume; Kanaley, Jill A; Vieira-Potter, Victoria J

2018-01-02

Loss of ovarian hormones is associated with increased adiposity, white adipose tissue (WAT) inflammation, and insulin resistance (IR). Previous work demonstrated ovariectomized (OVX) rats bred for high aerobic fitness (HCR) are protected against weight gain and IR compared to rats bred for low aerobic fitness (LCR) yet wheel running prevents OVX-induced IR in LCR rats. The purpose of this study was to determine whether adipose tissue immunometabolic characteristics from female HCR and LCR rats differs before or after OVX, and whether wheel running mitigates OVX-induced adipose tissue immunometabolic changes in LCR rats. Female OVX HCR and LCR rats were all fed a high fat diet (HFD) (n = 7-8/group) and randomized to either a running wheel or remain sedentary for 11 weeks. Ovary-intact rats (n = 7-12/group) were fed a standard chow diet with no wheel. Ovary-intact LCR rats had a greater visceral WAT inflammatory profile compared to HCR. Following OVX, sedentary LCR rats had greater serum leptin (p<0.001) and WAT inflammation (p<0.05) than sedentary HCR. Wheel running normalized the elevated serum leptin and reduced both visceral (p<0.05) and subcutaneous (p<0.03) WAT inflammatory markers in the LCR rats. Paradoxically, wheel running increased some markers of WAT inflammation in OVX HCR rats (p<0.05), which correlated with observed weight gain. Taken together, HCR rats appear to have a healthier WAT immune and metabolic profile compared to LCR, even following OVX. Wheel running improves WAT health in previously sedentary LCR rats. On the other hand, increased WAT inflammation is associated with adiposity gain despite a high volume of wheel running in HCR rats.

Glutathionylation of Yersinia pestis LcrV and Its Effects on Plague Pathogenesis

PubMed Central

Mitchell, Anthony; Tam, Christina; Elli, Derek; Charlton, Thomas; Osei-Owusu, Patrick; Fazlollahi, Farbod; Faull, Kym F.

2017-01-01

ABSTRACT Glutathionylation, the formation of reversible mixed disulfides between glutathione and protein cysteine residues, is a posttranslational modification previously observed for intracellular proteins of bacteria. Here we show that Yersinia pestis LcrV, a secreted protein capping the type III secretion machine, is glutathionylated at Cys273 and that this modification promotes association with host ribosomal protein S3 (RPS3), moderates Y. pestis type III effector transport and killing of macrophages, and enhances bubonic plague pathogenesis in mice and rats. Secreted LcrV was purified and analyzed by mass spectrometry to reveal glutathionylation, a modification that is abolished by the codon substitution Cys273Ala in lcrV. Moreover, the lcrVC273A mutation enhanced the survival of animals in models of bubonic plague. Investigating the molecular mechanism responsible for these virulence attributes, we identified macrophage RPS3 as a ligand of LcrV, an association that is perturbed by the Cys273Ala substitution. Furthermore, macrophages infected by the lcrVC273A variant displayed accelerated apoptotic death and diminished proinflammatory cytokine release. Deletion of gshB, which encodes glutathione synthetase of Y. pestis, resulted in undetectable levels of intracellular glutathione, and we used a Y. pestis ΔgshB mutant to characterize the biochemical pathway of LcrV glutathionylation, establishing that LcrV is modified after its transport to the type III needle via disulfide bond formation with extracellular oxidized glutathione. PMID:28512097

Intrinsic aerobic capacity impacts susceptibility to acute high-fat diet-induced hepatic steatosis.

PubMed

Morris, E Matthew; Jackman, Matthew R; Johnson, Ginger C; Liu, Tzu-Wen; Lopez, Jordan L; Kearney, Monica L; Fletcher, Justin A; Meers, Grace M E; Koch, Lauren G; Britton, Stephen L; Rector, R Scott; Ibdah, Jamal A; MacLean, Paul S; Thyfault, John P

2014-08-15

Aerobic capacity/fitness significantly impacts susceptibility for fatty liver and diabetes, but the mechanisms remain unknown. Herein, we utilized rats selectively bred for high (HCR) and low (LCR) intrinsic aerobic capacity to examine the mechanisms by which aerobic capacity impacts metabolic vulnerability for fatty liver following a 3-day high-fat diet (HFD). Indirect calorimetry assessment of energy metabolism combined with radiolabeled dietary food was employed to examine systemic metabolism in combination with ex vivo measurements of hepatic lipid oxidation. The LCR, but not HCR, displayed increased hepatic lipid accumulation in response to the HFD despite both groups increasing energy intake. However, LCR rats had a greater increase in energy intake and demonstrated greater daily weight gain and percent body fat due to HFD compared with HCR. Additionally, total energy expenditure was higher in the larger LCR. However, controlling for the difference in body weight, the LCR has lower resting energy expenditure compared with HCR. Importantly, respiratory quotient was significantly higher during the HFD in the LCR compared with HCR, suggesting reduced whole body lipid utilization in the LCR. This was confirmed by the observed lower whole body dietary fatty acid oxidation in LCR compared with HCR. Furthermore, LCR liver homogenate and isolated mitochondria showed lower complete fatty acid oxidation compared with HCR. We conclude that rats bred for low intrinsic aerobic capacity show greater susceptibility for dietary-induced hepatic steatosis, which is associated with a lower energy expenditure and reduced whole body and hepatic mitochondrial lipid oxidation.

LCR-eXXXplorer: a web platform to search, visualize and share data for low complexity regions in protein sequences.

PubMed

Kirmitzoglou, Ioannis; Promponas, Vasilis J

2015-07-01

Local compositionally biased and low complexity regions (LCRs) in amino acid sequences have initially attracted the interest of researchers due to their implication in generating artifacts in sequence database searches. There is accumulating evidence of the biological significance of LCRs both in physiological and in pathological situations. Nonetheless, LCR-related algorithms and tools have not gained wide appreciation across the research community, partly due to the fact that only a handful of user-friendly software is currently freely available. We developed LCR-eXXXplorer, an extensible online platform attempting to fill this gap. LCR-eXXXplorer offers tools for displaying LCRs from the UniProt/SwissProt knowledgebase, in combination with other relevant protein features, predicted or experimentally verified. Moreover, users may perform powerful queries against a custom designed sequence/LCR-centric database. We anticipate that LCR-eXXXplorer will be a useful starting point in research efforts for the elucidation of the structure, function and evolution of proteins with LCRs. LCR-eXXXplorer is freely available at the URL http://repeat.biol.ucy.ac.cy/lcr-exxxplorer. vprobon@ucy.ac.cy Supplementary data are available at Bioinformatics online. © The Author 2015. Published by Oxford University Press.

Variability of the protein sequences of lcrV between epidemic and atypical rhamnose-positive strains of Yersinia pestis.

PubMed

Anisimov, Andrey P; Panfertsev, Evgeniy A; Svetoch, Tat'yana E; Dentovskaya, Svetlana V

2007-01-01

Sequencing of lcrV genes and comparison of the deduced amino acid sequences from ten Y. pestis strains belonging mostly to the group of atypical rhamnose-positive isolates (non-pestis subspecies or pestoides group) showed that the LcrV proteins analyzed could be classified into five sequence types. This classification was based on major amino acid polymorphisms among LcrV proteins in the four "hot points" of the protein sequences. Some additional minor polymorphisms were found throughout these sequence types. The "hot points" corresponded to amino acids 18 (Lys --> Asn), 72 (Lys --> Arg), 273 (Cys --> Ser), and 324-326 (Ser-Gly-Lys --> Arg) in the LcrV sequence of the reference Y. pestis strain CO92. One possible explanation for polymorphism in amino acid sequences of LcrV among different strains is that strain-specific variation resulted from adaptation of the plague pathogen to different rodent and lagomorph hosts.

Adapting in vitro embryonic stem cell differentiation to the study of locus control regions.

PubMed

Lahiji, Armin; Kučerová-Levisohn, Martina; Holmes, Roxanne; Zúñiga-Pflücker, Juan Carlos; Ortiz, Benjamin D

2014-05-01

Numerous locus control region (LCR) activities have been discovered in gene loci important to immune cell development and function. LCRs are a distinct class of cis-acting gene regulatory elements that appear to contain all the DNA sequence information required to establish an independently and predictably regulated gene expression program at any genomic site in native chromatin of a whole animal. As such, LCR-regulated transgenic reporter systems provide invaluable opportunities to investigate the mechanisms of gene regulatory DNA action during development. Furthermore the qualities of LCR-driven gene expression, including spatiotemporal specificity and "integration site-independence" would be highly desirable to incorporate into vectors used in therapeutic genetic engineering. Thus, advancement in the methods used to investigate LCRs is of considerable basic and translational significance. We study the LCR present in the mouse T cell receptor (TCR)-α gene locus. Until recently, transgenic mice provided the only experimental model capable of supporting the entire spectrum of LCR activities. We have recently reported complete manifestation of TCRα LCR function in T cells derived in vitro from mouse embryonic stem cells (ESC), thus validating a complete cell culture model for the full range of LCR activities seen in transgenic mice. Here we discuss the critical parameters involved in studying LCR-regulated gene expression during in vitro hematopoietic differentiation from ESCs. This advance provides an approach to speed progress in the LCR field, and facilitate the clinical application of its findings, particularly to the genetic engineering of T cells. Copyright © 2014 Elsevier B.V. All rights reserved.

Platelet larger cell ratio (P-LCR) in patients with dyslipidemia.

PubMed

Grotto, H Z W; Noronha, J F A

2004-10-01

We have described preliminary results of platelet larger cell ratio (P-LCR) analysis, provided by an automated hematology analyzer in patients with hypercholesterolemia and/or hypertriglyceremia. P-LCR values were significantly higher in patients (n = 41) than in control normal group (n = 20) (P < 0.0001). Larger platelets are more reactive and contribute to vasooclusive events in patients with dyslipidemia. P-LCR may be used as an indicator of risk factor for thromboembolic ischemic events.

TRPV1 channels in the nucleus of the solitary tract mediate thermal prolongation of the LCR in decerebrate piglets

PubMed Central

Xia, Luxi; Bartlett, Donald; Leiter, J.C.

2011-01-01

Elevating body temperature or just the temperature of the dorsal medulla by approximately 2 °C prolongs the laryngeal chemoreflex (LCR) in decerebrate neonatal piglets. We tested the hypothesis that transient receptor potential vanilloid 1 (TRPV1) receptors in the nucleus of the solitary tract (NTS) mediate thermal prolongation of the LCR. We studied the effect of a selective TRPV1 receptor antagonist on thermal prolongation of the LCR, and we tested the effect of a TRPV1 agonist on the duration of the LCR under normothermic conditions. We studied 37 decerebrate neonatal piglets between the ages of post-natal days 4 and 7. The TRPV1 receptor antagonist, 5−iodoresiniferatoxin (65 microM/L in 100 nL), blocked thermal prolongation of the LCR when injected bilaterally into the region of the NTS. The TRPV1 agonist, resiniferatoxin (0.65-1.0 mM/L in 100 nL), prolonged the LCR after bilateral injection into the NTS even when the body temperature of each piglet was normal. The effect of the TRPV1 agonists could be blocked by treatment with the GABAA receptor antagonist, bicuculline, whether given intravenously (0.3 mg/kg) or focally injected bilaterally into the NTS (10 mM in 100nL). We conclude that TRPV1 receptors in the NTS mediate thermal prolongation of the LCR. PMID:21276877

Cooperativeness of the higher chromatin structure of the beta-globin locus revealed by the deletion mutations of DNase I hypersensitive site 3 of the LCR.

PubMed

Fang, Xiangdong; Xiang, Ping; Yin, Wenxuan; Stamatoyannopoulos, George; Li, Qiliang

2007-01-05

High-level transcription of the globin genes requires the enhancement by a distant element, the locus control region (LCR). Such long-range regulation in vivo involves spatial interaction between transcriptional elements, with intervening chromatin looping out. It has been proposed that the clustering of the HS sites of the LCR, the active globin genes, as well as the remote 5' hypersensitive sites (HSs) (HS-60/-62 in mouse, HS-110 in human) and 3'HS1 forms a specific spatial chromatin structure, termed active chromatin hub (ACH). Here we report the effects of the HS3 deletions of the LCR on the spatial chromatin structure of the beta-globin locus as revealed by the chromatin conformation capture (3C) technology. The small HS3 core deletion (0.23 kb), but not the large HS3 deletion (2.3 kb), disrupted the spatial interactions among all the HS sites of the LCR, the beta-globin gene and 3'HS1. We have previously demonstrated that the large HS3 deletion barely impairs the structure of the LCR holocomplex, while the structure is significantly disrupted by the HS3 core deletion. Taken together, these results suggest that the formation of the ACH is dependent on a largely intact LCR structure. We propose that the ACH indeed is an extension of the LCR holocomplex.

Hybrid capture-II and LCR-E7 PCR assays for HPV typing in cervical cytologic samples.

PubMed

Yamazaki, H; Sasagawa, T; Basha, W; Segawa, T; Inoue, M

2001-10-15

As part of an ongoing cohort study in the Hokuriku region of Japan, cervical cell samples from histologically confirmed normal (n = 114) or abnormal (n = 286) women were examined for the presence of HPV DNA using a second-generation hybrid capture assay (HCA-II) and LCR-E7 PCR. HCA-II detected low-risk (HPV-6, -11, -42, 43 and -44) and high-risk (HPV-16, -18, -31, -33, -35, -39, -45, -51, -52, -56, -58, -59 and -68) HPV types, while LCR-E7 PCR detected an additional 7 HPV types and some uncharacterized types. In screening of high-grade squamous intraepithelial lesions (HSILs) and invasive cervical cancer, the sensitivities of HCA-II and LCR-E7 PCR testing the high-risk HPV types were 83% and 81%, respectively, while the specificity of both assays was 93%. The sensitivity of LCR-E7 PCR increased to 87%, which was significantly higher than that in HCA-II, when testing both high-risk and other HPV types. Sixty-eight inconsistent results (17% of total tested) from HCA-II and LCR-E7 PCR were due to (i) low copy number of HPV genome (false-negative for HCA-II, 5.3% and for LCR-E7 PCR, 1.3%), (ii) infection with HPV types undetectable by HCA-II (4.8%), (iii) multiple HPV infections (5%) or (iv) unknown reasons (0.8%). LCR-E7 PCR revealed that infections with HPV-16, -18, -31, -33, -35, -51, -52, -56, -58 or -67 was a high risk for cancer since these types predominated in HSIL and invasive cervical cancer. Samples showing high relative light units (>20) with a high-risk probe in HCA-II also gave positive results in LCR-E7 PCR and were generally associated with abnormal cervical lesions. Thus, we propose that both HCA-II and LCR-E7 PCR are valuable screening tests for premalignant and malignant cervical lesions. Copyright 2001 Wiley-Liss, Inc.

The higher structure of chromatin in the LCR of the beta-globin locus changes during development.

PubMed

Fang, Xiangdong; Yin, Wenxuan; Xiang, Ping; Han, Hemei; Stamatoyannopoulos, George; Li, Qiliang

2009-11-27

The beta-globin locus control region (LCR) is able to enhance the expression of all globin genes throughout the course of development. However, the chromatin structure of the LCR at the different developmental stages is not well defined. We report DNase I and micrococcal nuclease hypersensitivity, chromatin immunoprecipitation analyses for histones H2A, H2B, H3, and H4, and 3C (chromatin conformation capture) assays of the normal and mutant beta-globin loci, which demonstrate that nucleosomes at the DNase I hypersensitive sites of the LCR could be either depleted or retained depending on the stages of development. Furthermore, MNase sensitivity and 3C assays suggest that the LCR chromatin is more open in embryonic erythroblasts than in definitive erythroblasts at the primary- and secondary-structure levels; however, the LCR chromatin is packaged more tightly in embryonic erythroblasts than in definitive erythroblasts at the tertiary chromatin level. Our study provides the first evidence that the occupancy of nucleosomes at a DNase I hypersensitive site is a developmental stage-related event and that embryonic and adult cells possess distinct chromatin structures of the LCR.

Demonstrations with an "LCR" Circuit

ERIC Educational Resources Information Center

Kraftmakher, Yaakov

2011-01-01

The "LCR" circuit is an important topic in the course of electricity and magnetism. Papers in this field consider mainly the forced oscillations and resonance. Our aim is to show how to demonstrate the free and self-excited oscillations in an "LCR" circuit. (Contains 4 figures.)

Relative Accuracy of Nucleic Acid Amplification Tests and Culture in Detecting Chlamydia in Asymptomatic Men

PubMed Central

Cheng, Hong; Macaluso, Maurizio; Vermund, Sten H.; Hook, Edward W.

2001-01-01

Published estimates of the sensitivity and specificity of PCR and ligase chain reaction (LCR) for detecting Chlamydia trachomatis are potentially biased because of study design limitations (confirmation of test results was limited to subjects who were PCR or LCR positive but culture negative). Relative measures of test accuracy are less prone to bias in incomplete study designs. We estimated the relative sensitivity (RSN) and relative false-positive rate (RFP) for PCR and LCR versus cell culture among 1,138 asymptomatic men and evaluated the potential bias of RSN and RFP estimates. PCR and LCR testing in urine were compared to culture of urethral specimens. Discordant results (PCR or LCR positive, but culture negative) were confirmed by using a sequence including the other DNA amplification test, direct fluorescent antibody testing, and a DNA amplification test to detect chlamydial major outer membrane protein. The RSN estimates for PCR and LCR were 1.45 (95% confidence interval [CI] = 1.3 to 1.7) and 1.49 (95% CI = 1.3 to 1.7), respectively, indicating that both methods are more sensitive than culture. Very few false-positive results were found, indicating that the specificity levels of PCR, LCR, and culture are high. The potential bias in RSN and RFP estimates were <5 and <20%, respectively. The estimation of bias is based on the most likely and probably conservative parameter settings. If the sensitivity of culture is between 60 and 65%, then the true sensitivity of PCR and LCR is between 90 and 97%. Our findings indicate that PCR and LCR are significantly more sensitive than culture, while the three tests have similar specificities. PMID:11682509

Human cancer risk estimation for 1,3-butadiene: An assessment of personal exposure and different microenvironments.

PubMed

Huy, Lai Nguyen; Lee, Shun Cheng; Zhang, Zhuozhi

2018-03-01

This study estimated the lifetime cancer risk (LCR) attributable to 1,3-butadiene (BD) personal exposure and to other microenvironments, including residential home, outdoor, in-office, in-vehicle, and dining. Detailed life expectancy by country (WHO), inhalation rate and body weight by gender reported by USEPA were used for the calculation, focusing on adult population (25≤Age<65). LCR estimation of the adult population due to personal exposure exceeded the USEPA benchmark of 1×10 -6 in many cities. For outdoor BD exposure, LCR estimations in 45 out of 175 cities/sites (sharing 26%) exceeded the USEPA benchmark. Out of the top 20 cities having high LCR estimations, developing countries contributed 19 cities, including 14, 3, 1, 1 cities in China, India, Chile, and Pakistan. One city in the United States was in the list due to the nearby industrial facilities. The LCR calculations for BD levels found in residential home, in-vehicle and dining microenvironments also exceeded 1×10 -6 in some cities, while LCR caused by in-office BD levels had the smallest risk. Four cities/regions were used for investigating source distributions to total LCR results because of their sufficient BD data. Home exposure contributed significantly to total LCR value (ranging 56% to 86%), followed by in-vehicle (4% to 38%) and dining (4 to 7%). Outdoor microenvironment shared highly in Tianjin with 6%, whereas in-office contributed from 2-3% for all cities. High LCR estimations found in developing countries highlighted the greater cancer risk caused by BD in other cities without available measurement data. Copyright © 2017 Elsevier B.V. All rights reserved.

Gut microbiota are linked to increased susceptibility to hepatic steatosis in low-aerobic-capacity rats fed an acute high-fat diet.

PubMed

Panasevich, Matthew R; Morris, E M; Chintapalli, S V; Wankhade, U D; Shankar, K; Britton, S L; Koch, L G; Thyfault, J P; Rector, R S

2016-07-01

Poor aerobic fitness is linked to nonalcoholic fatty liver disease and increased all-cause mortality. We previously found that rats with a low capacity for running (LCR) that were fed an acute high-fat diet (HFD; 45% kcal from fat) for 3 days resulted in positive energy balance and increased hepatic steatosis compared with rats that were highly aerobically fit with a high capacity for running (HCR). Here, we tested the hypothesis that poor physiological outcomes in LCR rats following acute HFD feeding are associated with alterations in cecal microbiota. LCR rats exhibited greater body weight, feeding efficiency, 3 days of body weight change, and liver triglycerides after acute HFD feeding compared with HCR rats. Furthermore, compared with HCR rats, LCR rats exhibited reduced expression of intestinal tight junction proteins. Cecal bacterial 16S rDNA revealed that LCR rats had reduced cecal Proteobacteria compared with HCR rats. Microbiota of HCR rats consisted of greater relative abundance of Desulfovibrionaceae and unassigned genera within this family, suggesting increased reduction of endogenous mucins and proteins. Although feeding rats an acute HFD led to reduced Firmicutes in both strains, short-chain fatty acid-producing Phascolarctobacterium was reduced in LCR rats. In addition, Ruminococcae and Ruminococcus were negatively correlated with energy intake in the LCR/HFD rats. Predicted metagenomic function suggested that LCR rats had a greater capacity to metabolize carbohydrate and energy compared with HCR rats. Overall, these data suggest that the populations and metabolic capacity of the microbiota in low-aerobically fit LCR rats may contribute to their susceptibility to acute HFD-induced hepatic steatosis and poor physiologic outcomes.

Gut microbiota are linked to increased susceptibility to hepatic steatosis in low-aerobic-capacity rats fed an acute high-fat diet

PubMed Central

Panasevich, Matthew R.; Morris, E. M.; Chintapalli, S. V.; Wankhade, U. D.; Shankar, K.; Britton, S. L.; Koch, L. G.; Thyfault, J. P.

2016-01-01

Poor aerobic fitness is linked to nonalcoholic fatty liver disease and increased all-cause mortality. We previously found that rats with a low capacity for running (LCR) that were fed an acute high-fat diet (HFD; 45% kcal from fat) for 3 days resulted in positive energy balance and increased hepatic steatosis compared with rats that were highly aerobically fit with a high capacity for running (HCR). Here, we tested the hypothesis that poor physiological outcomes in LCR rats following acute HFD feeding are associated with alterations in cecal microbiota. LCR rats exhibited greater body weight, feeding efficiency, 3 days of body weight change, and liver triglycerides after acute HFD feeding compared with HCR rats. Furthermore, compared with HCR rats, LCR rats exhibited reduced expression of intestinal tight junction proteins. Cecal bacterial 16S rDNA revealed that LCR rats had reduced cecal Proteobacteria compared with HCR rats. Microbiota of HCR rats consisted of greater relative abundance of Desulfovibrionaceae and unassigned genera within this family, suggesting increased reduction of endogenous mucins and proteins. Although feeding rats an acute HFD led to reduced Firmicutes in both strains, short-chain fatty acid-producing Phascolarctobacterium was reduced in LCR rats. In addition, Ruminococcae and Ruminococcus were negatively correlated with energy intake in the LCR/HFD rats. Predicted metagenomic function suggested that LCR rats had a greater capacity to metabolize carbohydrate and energy compared with HCR rats. Overall, these data suggest that the populations and metabolic capacity of the microbiota in low-aerobically fit LCR rats may contribute to their susceptibility to acute HFD-induced hepatic steatosis and poor physiologic outcomes. PMID:27288420

Characterization of nitrite degradation by Lactobacillus casei subsp. rhamnosus LCR 6013.

PubMed

Liu, Dong-mei; Wang, Pan; Zhang, Xin-yue; Xu, Xi-lin; Wu, Hui; Li, Li

2014-01-01

Nitrites are potential carcinogens. Therefore, limiting nitrites in food is critically important for food safety. The nitrite degradation capacity of Lactobacillus casei subsp. rhamnosus LCR 6013 was investigated in pickle fermentation. After LCR 6013 fermentation for 120 h at 37°C, the nitrite concentration in the fermentation system was significantly lower than that in the control sample without the LCR 6013 strain. The effects of NaCl and Vc on nitrite degradation by LCR 6013 in the De Man, Rogosa and Sharpe (MRS) medium were also investigated. The highest nitrite degradations, 9.29 mg/L and 9.89 mg/L, were observed when NaCl and Vc concentrations were 0.75% and 0.02%, respectively in the MRS medium, which was significantly higher than the control group (p ≤ 0.01). Electron capture/gas chromatography and indophenol blue staining were used to study the nitrite degradation pathway of LCR 6013. The nitrite degradation products contained N2O, but no NH4(+). The LCR 6013 strain completely degraded all NaNO2 (50.00 mg/L) after 16 h of fermentation. The enzyme activity of NiR in the periplasmic space was 2.5 times of that in the cytoplasm. Our results demonstrated that L. casei subsp. rhamnosus LCR 6013 can effectively degrade nitrites in both the pickle fermentation system and in MRS medium by NiR. Nitrites are degraded by the LCR 6013 strain, likely via the nitrate respiration pathway (NO2(-)>NO->N2O->N2), rather than the aammonium formation pathway (dissimilatory nitrate reduction to ammonium, DNRA), because the degradation products contain N2O, but not NH4(+).

Overweight female rats selectively breed for low aerobic capacity exhibit increased myocardial fibrosis and diastolic dysfunction

PubMed Central

Johnson, Megan S.; Ma, Lixin; Pulakat, Lakshmi; Mugerfeld, Irina; Hayden, Melvin R.; Garro, Mona; Knight, William; Britton, Steven L.; Koch, Lauren G.; Sowers, James R.

2012-01-01

The statistical association between endurance exercise capacity and cardiovascular disease suggests that impaired aerobic metabolism underlies the cardiovascular disease risk in men and women. To explore this connection, we applied divergent artificial selection in rats to develop low-capacity runner (LCR) and high-capacity runner (HCR) rats and found that disease risks segregated strongly with low running capacity. Here, we tested if inborn low aerobic capacity promotes differential sex-related cardiovascular effects. Compared with HCR males (HCR-M), LCR males (LCR-M) were overweight by 34% and had heavier retroperitoneal, epididymal, and omental fat pads; LCR females (LCR-F) were 20% heavier than HCR females (HCR-F), and their retroperitoneal, but not perireproductive or omental, fat pads were heavier as well. Unlike HCR-M, blood pressure was elevated in LCR-M, and this was accompanied by left ventricular (LV) hypertrophy. Like HCR-F, LCR-F exhibited normal blood pressure and LV weight as well as increased spontaneous cage activity compared with males. Despite normal blood pressures, LCR-F exhibited increased myocardial interstitial fibrosis and diastolic dysfunction, as indicated by increased LV stiffness, a decrease in the initial filling rate, and an increase in diastolic relaxation time. Although females exhibited increased arterial stiffness, ejection fraction was normal. Increased interstitial fibrosis and diastolic dysfunction in LCR-F was accompanied by the lowest protein levels of phosphorylated AMP-actived protein kinase [phospho-AMPK (Thr172)] and silent information regulator 1. Thus, the combination of risk factors, including female sex, intrinsic low aerobic capacity, and overweightness, promote myocardial stiffness/fibrosis sufficient to induce diastolic dysfunction in the absence of hypertension and LV hypertrophy. PMID:22345570

TRPV1 channels in the nucleus of the solitary tract mediate thermal prolongation of the LCR in decerebrate piglets.

PubMed

Xia, Luxi; Bartlett, Donald; Leiter, J C

2011-04-30

Elevating body temperature or just the temperature of the dorsal medulla by approximately 2°C prolongs the laryngeal chemoreflex (LCR) in decerebrate neonatal piglets. We tested the hypothesis that transient receptor potential vanilloid 1 (TRPV1) receptors in the nucleus of the solitary tract (NTS) mediate thermal prolongation of the LCR. We studied the effect of a selective TRPV1 receptor antagonist on thermal prolongation of the LCR, and we tested the effect of a TRPV1 agonist on the duration of the LCR under normothermic conditions. We studied 37 decerebrate neonatal piglets between the ages of post-natal days 4 and 7. The TRPV1 receptor antagonist, 5'-iodoresiniferatoxin (65μM/L in 100nL), blocked thermal prolongation of the LCR when injected bilaterally into the region of the NTS. The TRPV1 agonist, resiniferatoxin (0.65-1.0mM/L in 100nL), prolonged the LCR after bilateral injection into the NTS even when the body temperature of each piglet was normal. The effect of the TRPV1 agonists could be blocked by treatment with the GABA(A) receptor antagonist, bicuculline, whether given intravenously (0.3mg/kg) or focally injected bilaterally into the NTS (10mM in 100nL). We conclude that TRPV1 receptors in the NTS mediate thermal prolongation of the LCR. Copyright © 2011 Elsevier B.V. All rights reserved.

A light-colored region of caudal fin: a niche of melanocyte progenitors in crucian carp (Cyprinus carpio L.).

PubMed

Huang, Yaping; Luo, Yurong; Liu, Jinhui; Gui, Saiyu; Wang, Mei; Liu, Wenbin; Peng, Liangyue; Xiao, Yamei

2017-01-01

Melanocyte stem cells are a population of immature cells which sustain the self-renewal and replenish the differentiated melanocytes. In this research, a light-colored region (LCR) is observed at the heel of caudal fin in juvenile crucian carp. By cutting off the caudal fin, the operated caudal fin can regenerate in accordance with the original pigment pattern from the retained LCR. As markers of stem cells, Oct4 and Sox2 have been found to be highly expressed in the LCR as well as Mitfa, a label of the melanoblasts. In vitro, Mitfa + melanoblasts are observed in the cells which are derived from the LCR and transfected with Mitfa-EGFP reporter by using Tol2 transposon system. Furthermore, by real-time qPCR, it is shown that the level of sox2 mRNA is gradually decreased from the LCR to proximal and distal caudal fin, and that of mitfa mRNA in the proximal caudal fin (PCF) is higher than that in the LCR, while it is the lowest in the distal caudal fin. Hence, we propose that the LCR is a pigment progenitor niche, sending melanocytes to the distal of caudal fin, which gradually emerges as caudal fin grow. We reveal that the LCR of caudal fin might be a niche of pigment progenitors, and contribute to pigment-producing stem cells in crucian carp. © 2016 International Federation for Cell Biology.

SOX2 as a New Regulator of HPV16 Transcription.

PubMed

Martínez-Ramírez, Imelda; Del-Castillo-Falconi, Víctor; Mitre-Aguilar, Irma B; Amador-Molina, Alfredo; Carrillo-García, Adela; Langley, Elizabeth; Zentella-Dehesa, Alejandro; Soto-Reyes, Ernesto; García-Carrancá, Alejandro; Herrera, Luis A; Lizano, Marcela

2017-07-05

Persistent infections with high-risk human papillomavirus (HPV) constitute the main risk factor for cervical cancer development. HPV16 is the most frequent type associated to squamous cell carcinomas (SCC), followed by HPV18. The long control region (LCR) in the HPV genome contains the replication origin and sequences recognized by cellular transcription factors (TFs) controlling viral transcription. Altered expression of E6 and E7 viral oncogenes, modulated by the LCR, causes modifications in cellular pathways such as proliferation, leading to malignant transformation. The aim of this study was to identify specific TFs that could contribute to the modulation of high-risk HPV transcriptional activity, related to the cellular histological origin. We identified sex determining region Y (SRY)-box 2 (SOX2) response elements present in HPV16-LCR. SOX2 binding to the LCR was demonstrated by in vivo and in vitro assays. The overexpression of this TF repressed HPV16-LCR transcriptional activity, as shown through reporter plasmid assays and by the down-regulation of endogenous HPV oncogenes. Site-directed mutagenesis revealed that three putative SOX2 binding sites are involved in the repression of the LCR activity. We propose that SOX2 acts as a transcriptional repressor of HPV16-LCR, decreasing the expression of E6 and E7 oncogenes in a SCC context.

Aberrant expression of IFN-γ in Th2 cells from Th2 LCR-deficient mice.

PubMed

Hwang, Soo Seok; Kim, Kiwan; Lee, Wonyong; Lee, Gap Ryol

2012-08-03

The Th2 locus control region (LCR) has been shown to be a crucial cis-acting element for Th2 cytokine expression and Th2 cell differentiation. To study the role of Th2 LCR in ifng locus regulation, we examined the expression of IFN-γ in Th2 cells from Th2 LCR-deficient mice. We found IFN-γ to be aberrantly up-regulated. In addition, histone 3(H3)-acetylation and histone 3 lysine 4 (H3-K4)-methylation greatly increased at the ifng locus of the Th2 cells. GATA-3 and STAT6 bound to the ifng promoter in Th2 cells from the wild type but not from the Th2 LCR-deficient mice, and they directly repressed ifng expression in transient reporter assay. Moreover, ectopic expression of GATA-3 and STAT6-VT repressed the aberrant expression of the ifng gene and restored repressive chromatin state at the ifng locus in Th2 cells from Th2 LCR-deficient mice. These results suggest that expression of the ifng gene and chromatin remodeling of the ifng locus are under the control of a Th2 LCR-mediated Th2 differentiation program. Copyright © 2012 Elsevier Inc. All rights reserved.

Interleukin-1β and interleukin-6 enhance thermal prolongation of the LCR in decerebrate piglets.

PubMed

Xia, Luxi; Bartlett, Donald; Leiter, J C

2016-08-01

Thermal stress and prior upper respiratory tract infection are risk factors for the Sudden Infant Death Syndrome. The adverse effects of prior infection are likely mediated by interleukin-1β (IL-1β). Therefore, we examined the single and combined effects of IL-1β and elevated body temperature on the duration of the Laryngeal Chemoreflex (LCR) in decerebrate neonatal piglets ranging in age from post-natal day (P) 3 to P7. We examined the effects of intraperitoneal (I.P.) injections of 0.3mg/Kg IL-1β with or without I.P. 10mg/Kg indomethacin pretreatment on the duration of the LCR, and in the same animals we also examined the duration of the LCR when body temperature was elevated approximately 2°C. We found that IL-1β significantly increased the duration of the LCR even when body temperature was held constant. There was a significant multiplicative effect when elevated body temperature was combined with IL-1β treatment: prolongation of the LCR was significantly greater than the sum of independent thermal and IL-1β-induced prolongations of the LCR. The effects of IL-1β, but not elevated body temperature, were blocked by pretreatment with indomethacin alone. We also tested the interaction between IL-6 given directly into the nucleus of the solitary tract (NTS) bilaterally in 100ngm microinjections of 50μL and pretreatment with indomethacin. Here again, there was a multiplicative effect of IL-6 treatment and elevated body temperature, which significantly prolonged the LCR. The effect of IL-6 on the LCR, but not elevated body temperature, was blocked by pretreatment with indomethacin. We conclude that cytokines interact with elevated body temperature, probably through direct thermal effects on TRPV1 receptors expressed pre-synaptically in the NTS and through cytokine-dependent sensitization of the TRPV1 receptor. This sensitization is likely initiated by cyclo-oxygenase-2 dependent synthesis of prostaglandin E2, which is stimulated by elevated levels of IL-1β or IL-6. Inflammatory sensitization of the LCR coupled with thermal prolongation of the LCR may increase the propensity for apnea and Sudden Infant Death Syndrome. Copyright © 2016 Elsevier B.V. All rights reserved.

Interleukin-1β and interleukin-6 enhance thermal prolongation of the LCR in decerebrate piglets

PubMed Central

Xia, Luxi; Bartlett, Donald; Leiter, J.C.

2017-01-01

Thermal stress and prior upper respiratory tract infection are risk factors for the Sudden Infant Death Syndrome. The adverse effects of prior infection are likely mediated by interleukin-1β (IL-1β). Therefore, we examined the single and combined effects of IL-1β and elevated body temperature on the duration of the Laryngeal Chemoreflex (LCR) in decerebrate neonatal piglets ranging in age from post-natal day (P) 3 to P7. We examined the effects of intraperitoneal (I.P.) injections of 0.3 mg/Kg IL-1β with or without I.P. 10 mg/Kg indomethacin pretreatment on the duration of the LCR, and in the same animals we also examined the duration of the LCR when body temperature was elevated approximately 2 °C. We found that IL-1β significantly increased the duration of the LCR even when body temperature was held constant. There was a significant multiplicative effect when elevated body temperature was combined with IL-1β treatment: prolongation of the LCR was significantly greater than the sum of independent thermal and IL-1β-induced prolongations of the LCR. The effects of IL-1β, but not elevated body temperature, were blocked by pretreatment with indomethacin alone. We also tested the interaction between IL-6 given directly into the nucleus of the solitary tract (NTS) bilaterally in 100 ngm microinjections of 50 μL and pre-treatment with indomethacin. Here again, there was a multiplicative effect of IL-6 treatment and elevated body temperature, which significantly prolonged the LCR. The effect of IL-6 on the LCR, but not elevated body temperature, was blocked by pretreatment with indomethacin. We conclude that cytokines interact with elevated body temperature, probably through direct thermal effects on TRPV1 receptors expressed pre-synaptically in the NTS and through cytokine-dependent sensitization of the TRPV1 receptor. This sensitization is likely initiated by cyclo-oxygenase-2 dependent synthesis of prostaglandin E2, which is stimulated by elevated levels of IL-1β or IL-6. Inflammatory sensitization of the LCR coupled with thermal prolongation of the LCR may increase the propensity for apnea and Sudden Infant Death Syndrome. PMID:27181326

Glutathionylation of Yersinia pestis LcrV and Its Effects on Plague Pathogenesis.

PubMed

Mitchell, Anthony; Tam, Christina; Elli, Derek; Charlton, Thomas; Osei-Owusu, Patrick; Fazlollahi, Farbod; Faull, Kym F; Schneewind, Olaf

2017-05-16

Glutathionylation, the formation of reversible mixed disulfides between glutathione and protein cysteine residues, is a posttranslational modification previously observed for intracellular proteins of bacteria. Here we show that Yersinia pestis LcrV, a secreted protein capping the type III secretion machine, is glutathionylated at Cys 273 and that this modification promotes association with host ribosomal protein S3 (RPS3), moderates Y. pestis type III effector transport and killing of macrophages, and enhances bubonic plague pathogenesis in mice and rats. Secreted LcrV was purified and analyzed by mass spectrometry to reveal glutathionylation, a modification that is abolished by the codon substitution Cys 273 Ala in lcrV Moreover, the lcrV C273A mutation enhanced the survival of animals in models of bubonic plague. Investigating the molecular mechanism responsible for these virulence attributes, we identified macrophage RPS3 as a ligand of LcrV, an association that is perturbed by the Cys 273 Ala substitution. Furthermore, macrophages infected by the lcrV C273A variant displayed accelerated apoptotic death and diminished proinflammatory cytokine release. Deletion of gshB , which encodes glutathione synthetase of Y. pestis , resulted in undetectable levels of intracellular glutathione, and we used a Y. pestis Δ gshB mutant to characterize the biochemical pathway of LcrV glutathionylation, establishing that LcrV is modified after its transport to the type III needle via disulfide bond formation with extracellular oxidized glutathione. IMPORTANCE Yersinia pestis , the causative agent of plague, has killed large segments of the human population; however, the molecular bases for the extraordinary virulence attributes of this pathogen are not well understood. We show here that LcrV, the cap protein of bacterial type III secretion needles, is modified by host glutathione and that this modification contributes to the high virulence of Y. pestis in mouse and rat models for bubonic plague. These data suggest that Y. pestis exploits glutathione in host tissues to activate a virulence strategy, thereby accelerating plague pathogenesis. Copyright © 2017 Mitchell et al.

LCR testing for gonorrhoea and chlamydia in population surveys and other screenings of low prevalence populations: coping with decreased positive predictive value.

PubMed

Zenilman, J M; Miller, W C; Gaydos, C; Rogers, S M; Turner, C F

2003-04-01

Nucleic acid amplification tests have facilitated field based STD studies and increased screening activities. However, even with highly specific tests, the positive predictive value (PPV) of such tests may be lower than desirable in low prevalence populations. We estimated PPVs for a single LCR test in a population survey in which positive specimens were retested. The Baltimore STD and Behavior Survey (BSBS) was a population based behavioural survey of adults which included collecting urine specimens to assess the prevalence of gonorrhoea and chlamydial infection. Gonorrhoea and chlamydial infection were diagnosed by ligase chain reaction (LCR). Nearly all positive results were retested by LCR. Because of cost considerations, negative results were not confirmed. Predicted curves for the PPV were calculated for a single testing assuming an LCR test sensitivity of 95%, and test specificities in the range 95.0%-99.9%, for disease prevalences between 1% and 10%. Positive specimens were retested to derive empirical estimates of the PPV of a positive result on a single LCR test. 579 participants age 18-35 provided urine specimens. 20 (3.5%) subjects initially tested positive for chlamydial infection, and 39 (6.7%) tested positive for gonococcal infection. If positive results on the repeat LCR are taken as confirmation of a "true" infection, the observed PPV for the first LCR testing was 89.5% for chlamydial infection and 83.3% for gonorrhoea. This is within the range of theoretical PPVs calculated from the assumed sensitivities and specificities of the LCR assays. Empirical performance of a single LCR testing approximated the theoretically predicted PPV in this field study. This result demonstrates the need to take account of the lower PPVs obtained when such tests are used in field studies or clinical screening of low prevalence populations. Repeat testing of specimens, preferably with a different assay (for example, polymerase chain reaction), and disclosure of the non-trivial potential for false positive test results would seem appropriate in all such studies.

Effects of Methylation Status of CpG Sites within the HPV16 Long Control Region on HPV16-Positive Head and Neck Cancer Cells.

PubMed

Zhang, Chunlin; Deng, Zeyi; Pan, Xiaoli; Uehara, Takayuki; Suzuki, Mikio; Xie, Minqiang

2015-01-01

To map comprehensively the methylation status of the CpG sites within the HPV16 long control region (LCR) in HPV-positive cancer cells, and to explore further the effects of methylation status of HPV16 LCR on cell bioactivity and E6 and E7 expression. In addition, to analyze the methylation status of the LCR in HPV-positive oropharyngeal squamous cell carcinoma (OPSCC) patients. Methylation patterns of HPV16 LCR in UM-SCC47, CaSki, and SiHa cells and HPV16-positiive OPSCC specimens were detected by bisulfite-sequencing PCR and TA cloning. For cells treated with 5-aza-2'-deoxycytidine and E6 and E7 knockdown, MTS and trypan blue staining, annexin-V and 7-AAD staining, and prodidium iodide were used to evaluate cell growth and cell proliferation, cell apoptosis, and cell cycle arrest, respectively. E6 and E7 mRNA and protein expression were analyzed by quantitative real-time PCR and immunocytochemistry, respectively. Hypermethylation status of the LCR in UM-SCC47 (79.8%) and CaSki cells (90.0%) and unmethylation status of the LCR in SiHa cells (0%) were observed. Upon demethylation, the cells with different methylation levels responded differently during growth, apoptosis, and cell cycle arrest, as well as in terms of their E6 and E7 expression. In HPV16-positive OPSCC patients, the methylation rates were 9.5% in the entire LCR region, 13.9% in the 5'-LCR, 6.0% in the E6 enhancer, and 9.5% in the p97 promoter, and hypermethylation of p97 promoter was found in a subset of cases (20.0%, 2/10). Our study revealed two different methylation levels of the LCR in HPV16-positive cancer cells and OPSCC patients, which may represent different carcinogenesis mechanisms of HPV-positive cancers cells. Demethylating the meCpGs in HPV16 LCR might be a potential target for a subgroup of HPV16-positive patients with head and neck squamous cell carcinoma.

Brain diabetic neurodegeneration segregates with low intrinsic aerobic capacity

PubMed Central

Choi, Joungil; Chandrasekaran, Krish; Demarest, Tyler G; Kristian, Tibor; Xu, Su; Vijaykumar, Kadambari; Dsouza, Kevin Geoffrey; Qi, Nathan R; Yarowsky, Paul J; Gallipoli, Rao; Koch, Lauren G; Fiskum, Gary M; Britton, Steven L; Russell, James W

2014-01-01

Objectives Diabetes leads to cognitive impairment and is associated with age-related neurodegenerative diseases including Alzheimer's disease (AD). Thus, understanding diabetes-induced alterations in brain function is important for developing early interventions for neurodegeneration. Low-capacity runner (LCR) rats are obese and manifest metabolic risk factors resembling human “impaired glucose tolerance” or metabolic syndrome. We examined hippocampal function in aged LCR rats compared to their high-capacity runner (HCR) rat counterparts. Methods Hippocampal function was examined using proton magnetic resonance spectroscopy and imaging, unbiased stereology analysis, and a Y maze. Changes in the mitochondrial respiratory chain function and levels of hyperphosphorylated tau and mitochondrial transcriptional regulators were examined. Results The levels of glutamate, myo-inositol, taurine, and choline-containing compounds were significantly increased in the aged LCR rats. We observed a significant loss of hippocampal neurons and impaired cognitive function in aged LCR rats. Respiratory chain function and activity were significantly decreased in the aged LCR rats. Hyperphosphorylated tau was accumulated within mitochondria and peroxisome proliferator-activated receptor-gamma coactivator 1α, the NAD+-dependent protein deacetylase sirtuin 1, and mitochondrial transcription factor A were downregulated in the aged LCR rat hippocampus. Interpretation These data provide evidence of a neurodegenerative process in the hippocampus of aged LCR rats, consistent with those seen in aged-related dementing illnesses such as AD in humans. The metabolic and mitochondrial abnormalities observed in LCR rat hippocampus are similar to well-described mechanisms that lead to diabetic neuropathy and may provide an important link between cognitive and metabolic dysfunction. PMID:25356430

Physically active rats lose more weight during calorie restriction.

PubMed

Smyers, Mark E; Bachir, Kailey Z; Britton, Steven L; Koch, Lauren G; Novak, Colleen M

2015-02-01

Daily physical activity shows substantial inter-individual variation, and low physical activity is associated with obesity and weight gain. Elevated physical activity is also associated with high intrinsic aerobic capacity, which confers considerable metabolic health benefits. Rats artificially selected for high intrinsic aerobic capacity (high-capacity runners, HCR) are more physically active than their low-capacity counterparts (low-capacity runners, LCR). To test the hypothesis that physical activity counters metabolic thriftiness, we measured physical activity and weight loss during three weeks of 50% calorie restriction (CR) in the HCR and LCR rat lines. At baseline, HCR ate more and were more active than LCR; this was seen in male rats, where LCR are considerably heavier than HCR, as well as in a set of female rats where body weight did not differ between the lines, demonstrating that this effect is consistent across sex and not secondary to body weight. We show for the first time that HCR lose more weight than LCR relative to baseline. Physical activity levels declined throughout CR, and this was more pronounced in HCR than in LCR, yet some aspects of activity remained elevated in HCR relative to LCR even during CR. This is consistent with the idea that low physical activity contributes to metabolic thriftiness during food restriction, allowing LCR to defend body mass, particularly lean mass. This has implications for physical activity during diet-induced weight loss, the genetic underpinnings of individual differences in weight loss during a diet, and the potential evolutionary opposition between metabolic thriftiness and aerobic capacity. Copyright © 2014 Elsevier Inc. All rights reserved.

Synergistic and additive properties of the beta-globin locus control region (LCR) revealed by 5'HS3 deletion mutations: implication for LCR chromatin architecture.

PubMed

Fang, Xiangdong; Sun, Jin; Xiang, Ping; Yu, Man; Navas, Patrick A; Peterson, Kenneth R; Stamatoyannopoulos, George; Li, Qiliang

2005-08-01

Deletion of the 234-bp core element of the DNase I hypersensitive site 3 (5'HS3) of the locus control region (LCR) in the context of a human beta-globin locus yeast artificial chromosome (beta-YAC) results in profound effects on globin gene expression in transgenic mice. In contrast, deletion of a 2.3-kb 5'HS3 region, which includes the 234-bp core sequence, has a much milder phenotype. Here we report the effects of these deletions on chromatin structure in the beta-globin locus of adult erythroblasts. The 234-bp 5'HS3 deletion abolished histone acetylation throughout the beta-globin locus; recruitment of RNA polymerase II (pol II) to the LCR and beta-globin gene promoter was reduced to a basal level; and formation of all the 5' DNase I hypersensitive sites of the LCR was disrupted. The 2.3-kb 5'HS3 deletion mildly reduced the level of histone acetylation but did not change the profile across the whole locus; the 5' DNase I hypersensitive sites of the LCR were formed, but to a lesser extent; and recruitment of pol II was reduced, but only marginally. These data support the hypothesis that the LCR forms a specific chromatin structure and acts as a single entity. Based on these results we elaborate on a model of LCR chromatin architecture which accommodates the distinct phenotypes of the 5'HS3 and HS3 core deletions.

Physically Active Rats Lose More Weight during Calorie Restriction

PubMed Central

Smyers, Mark E.; Bachir, Kailey Z.; Britton, Steven L.; Koch, Lauren G.; Novak, Colleen M.

2014-01-01

Daily physical activity shows substantial inter-individual variation, and low physical activity is associated with obesity and weight gain. Elevated physical activity is also associated with high intrinsic aerobic capacity, which confers considerable metabolic health benefits. Rats artificially selected for high intrinsic aerobic capacity (high-capacity runners, HCR) are more physically active than their low-capacity counterparts (low-capacity runners, LCR). To test the hypothesis that physical activity counters metabolic thriftiness, we measured physical activity and weight loss during three weeks of 50% calorie restriction (CR) in the HCR and LCR rat lines. At baseline, HCR ate more and were more active than LCR; this was seen in male rats, where LCR are considerably heavier than HCR, as well as in a set of female rats where body weight did not differ between the lines, demonstrating that this effect is consistent across sex and not secondary to body weight. We show for the first time that HCR lose more weight than LCR relative to baseline. Physical activity levels declined throughout CR, and this was more pronounced in HCR than in LCR, yet some aspects of activity remained elevated in HCR relative to LCR even during CR. This is consistent with the idea that low physical activity contributes to metabolic thriftiness during food restriction, allowing LCR to defend body mass, particularly lean mass. This has implications for physical activity during diet-induced weight loss, the genetic underpinnings of individual differences in weight loss during a diet, and the potential evolutionary opposition between metabolic thriftiness and aerobic capacity. PMID:25449411

Plasmid-determined cytotoxicity in Yersinia pestis and Yersinia pseudotuberculosis.

PubMed Central

Goguen, J D; Walker, W S; Hatch, T P; Yother, J

1986-01-01

Yersinia pestis KIM5 was found to be cytotoxic for the IC21 and P388D1 mouse macrophage cell lines, as well as for resident peritoneal macrophages from C57BL/6 mice. Affected cells phagocytosed KIM5 inefficiently, became spherical, detached readily from culture dishes, and retained 51Cr poorly. The cytotoxic effect was dependent on the presence of the 75-kilobase plasmid pCD1. Because this plasmid also encodes the low calcium response (LCR), three Mu d1 insertion mutants previously shown to be LCR- and of reduced virulence in mice were examined for cytotoxicity; all were found to be atoxic. The insertions in these mutants lie within three distinct LCR loci (lcrB, C, and D). Like LCR, cytotoxicity was expressed only at 37 degrees C. Unlike LCR, it was not influenced by Ca2+ concentration, indicating that the V and W antigens are probably not involved. Yersinia pseudotuberculosis was found to have a similar plasmid-dependent cytotoxicity. Thus, biological activity observed as cytotoxicity in vitro may well be a common feature contributing to virulence of the yersiniae. Images PMID:3949380

Intranasal delivery of a protein subunit vaccine using a Tobacco Mosaic Virus platform protects against pneumonic plague.

PubMed

Arnaboldi, Paul M; Sambir, Mariya; D'Arco, Christina; Peters, Lauren A; Seegers, Jos F M L; Mayer, Lloyd; McCormick, Alison A; Dattwyler, Raymond J

2016-11-11

Yersinia pestis, one of history's deadliest pathogens, has killed millions over the course of human history. It has attributes that make it an ideal choice to produce mass casualties and is a prime candidate for use as a biological weapon. When aerosolized, Y. pestis causes pneumonic plague, a pneumonia that is 100% lethal if not promptly treated with effective antibiotics. Currently, there is no FDA approved plague vaccine. The current lead vaccine candidate, a parenterally administered protein subunit vaccine comprised of the Y. pestis virulence factors, F1 and LcrV, demonstrated variable levels of protection in primate pneumonic plague models. As the most likely mode of exposure in biological attack with Y. pestis is by aerosol, this raises a question of whether this parenteral vaccine will adequately protect humans against pneumonic plague. In the present study we evaluated two distinct mucosal delivery platforms for the intranasal (IN) administration of LcrV and F1 vaccine proteins, a live bacterial vector, Lactobacillus plantarum, and a Tobacco Mosaic Virus (TMV) based delivery platform. IN administration of L. plantarum expressing LcrV, or TMV-conjugated to LcrV and F1 (TMV-LcrV+TMV-F1) resulted in the similar induction of high titers of IgG antibodies and evidence of proinflammatory cytokine secretion. However, only the TMV-conjugate delivery platform protected against subsequent lethal challenge with Y. pestis. TMV-LcrV+TMV-F1 co-vaccinated mice had no discernable morbidity and no mortality, while mice vaccinated with L. plantarum expressing LcrV or rLcrV+rF1 without TMV succumbed to infection or were only partially protected. Thus, TMV is a suitable mucosal delivery platform for an F1-LcrV subunit vaccine that induces complete protection against pneumonic infection with a lethal dose of Y. pestis in mice. Copyright © 2016 Elsevier Ltd. All rights reserved.

Parasitism and body condition in humpback chub from the Colorado and Little Colorado Rivers, Grand Canyon, Arizona

USGS Publications Warehouse

Hoffnagle, Timothy L.; Choudhury, Anindo; Cole, Rebecca A.

2006-01-01

Glen Canyon Dam has greatly altered the Colorado River in Grand Canyon. The Little Colorado River (LCR) provides a small refuge of seasonally warm and turbid water that is thought to be more suitable than the Colorado River for endangered humpback chub Gila cypha. However, the LCR has low productivity and contains nonnative fishes and parasites, which pose a threat to humpback chub. The Colorado River hosts a different suite of nonnative fishes and is cold and clear but more productive. We compared condition factor (K), abdominal fat index (AFI), and presence and number of two introduced pathogenic parasites (Lernaea cyprinacea and Bothriocephalus acheilognathi) between juvenile (<150 mm total length) humpback chub from the LCR and those from the Colorado River during 1996a??1999. Both K and AFI were lower and L. cyprinacea prevalence and B. acheilognathi prevalence were higher in LCR fish than in Colorado River fish for all years. Mean K and AFI were 0.622 and 0.48, respectively, in the LCR and 0.735 and 2.02, respectively, in the Colorado River, indicating that fish in the Colorado River were more robust. Mean prevalence of L. cyprinacea was 23.9% and mean intensity was 1.73 L. cyprinacea/infected fish in the LCR, whereas prevalence was 3.2% and intensity was 1.0 L. cyprinacea/infected fish in the Colorado River. Mean prevalence of B. acheilognathi was 51.0% and mean intensity was 25.0 B. acheilognathi/infected fish in the LCR, whereas prevalence was 15.8% and intensity was 12.0 B. acheilognathi/infected fish in the Colorado River. Increased parasitism and poorer body condition in humpback chub from the LCR challenge the paradigm that warmer LCR waters are more suitable for humpback chub than the colder Colorado River and indicate the need to consider the importance and benefits of all available habitats, as well as biotic and abiotic factors, when managing endangered species and their environment.

Flanking HS-62.5 and 3' HS1, and regions upstream of the LCR, are not required for beta-globin transcription.

PubMed

Bender, M A; Byron, Rachel; Ragoczy, Tobias; Telling, Agnes; Bulger, Michael; Groudine, Mark

2006-08-15

The locus control region (LCR) was thought to be necessary and sufficient for establishing and maintaining an open beta-globin locus chromatin domain in the repressive environment of the developing erythrocyte. However, deletion of the LCR from the endogenous locus had no significant effect on chromatin structure and did not silence transcription. Thus, the cis-regulatory elements that confer the open domain remain unidentified. The conserved DNaseI hypersensitivity sites (HSs) HS-62.5 and 3'HS1 that flank the locus, and the region upstream of the LCR have been implicated in globin gene regulation. The flanking HSs bind CCCTC binding factor (CTCF) and are thought to interact with the LCR to form a "chromatin hub" involved in beta-globin gene activation. Hispanic thalassemia, a deletion of the LCR and 27 kb upstream, leads to heterochromatinization and silencing of the locus. Thus, the region upstream of the LCR deleted in Hispanic thalassemia (upstream Hispanic region [UHR]) may be required for expression. To determine the importance of the UHR and flanking HSs for beta-globin expression, we generated and analyzed mice with targeted deletions of these elements. We demonstrate deletion of these regions alone, and in combination, do not affect transcription, bringing into question current models for the regulation of the beta-globin locus.

Ovariectomized Highly Fit Rats Are Protected against Diet-Induced Insulin Resistance.

PubMed

Park, Young-Min; Kanaley, Jill A; Zidon, Terese M; Welly, Rebecca J; Scroggins, Rebecca J; Britton, Steven L; Koch, Lauren G; Thyfault, John P; Booth, Frank W; Padilla, Jaume; Vieira-Potter, Victoria J

2016-07-01

In the absence of exercise training, rats selectively bred for high intrinsic aerobic capacity (high-capacity running (HCR)) are protected against ovariectomy (OVX)-induced insulin resistance (IR) and obesity compared with those bred for low intrinsic aerobic capacity (low-capacity running (LCR)). This study determined whether OVX HCR rats remain protected with exposure to high-fat diet (HFD) compared with OVX LCR rats. Female HCR and LCR rats (n = 36; age, 27-33 wk) underwent OVX and were randomized to a standard chow diet (NC, 5% kcal fat) or HFD (45% kcal fat) ad libitum for 11 wk. Total energy expenditure, resting energy expenditure, spontaneous physical activity (SPA), and glucose tolerance were assessed midway, whereas fasting circulating metabolic markers, body composition, adipose tissue distribution, and skeletal muscle adenosine monophosphate-activated protein kinase (AMPK), and mitochondrial markers were assessed at sacrifice. Both HCR and LCR rats experienced HFD-induced increases in total and visceral adiposity after OVX. Despite similar gains in adiposity, HCR rats were protected from HFD-induced IR and reduced total energy expenditure observed in LCR rats (P < 0.05). This metabolic protection was likely attributed to a compensatory increase in SPA and associated preservation of skeletal muscle AMPK activity in HCR; however, HFD significantly reduced SPA and AMPK activity in LCR (P < 0.05). In both lines, HFD reduced citrate synthase activity, gene expression of markers of mitochondrial biogenesis (tFAM, NRF1, and PGC-1α), and protein levels of mitochondrial oxidative phosphorylation complexes I, II, IV, and V in skeletal muscle (all P < 0.05). After OVX, HCR and LCR rats differentially respond to HFD such that HCR increase while LCR decrease SPA. This "physical activity compensation" likely confers protection from HFD-induced IR and reduced energy expenditure in HCR rats.

Human HMG box transcription factor HBP1: a role in hCD2 LCR function.

PubMed Central

Zhuma, T; Tyrrell, R; Sekkali, B; Skavdis, G; Saveliev, A; Tolaini, M; Roderick, K; Norton, T; Smerdon, S; Sedgwick, S; Festenstein, R; Kioussis, D

1999-01-01

The locus control region (LCR) of the human CD2 gene (hCD2) confers T cell-specific, copy-dependent and position-independent gene expression in transgenic mice. This LCR consists of a strong T cell-specific enhancer and an element without enhancer activity (designated HSS3), which is required for prevention of position effect variegation (PEV) in transgenic mice. Here, we identified the HMG box containing protein-1 (HBP1) as a factor binding to HSS3 of the hCD2 LCR. Within the LCR, HBP1 binds to a novel TTCATTCATTCA sequence that is higher in affinity than other recently reported HBP1-binding sites. Mice transgenic for a hCD2 LCR construct carrying a deletion of the HBP1-binding sequences show a propensity for PEV if the transgene integrates in a heterochromatic region of the chromosome such as the centromere or telomere. We propose that HBP1 plays an important role in chromatin opening and remodelling activities by binding to and bending the DNA, thus allowing DNA-protein and/or protein-protein interactions, which increase the probability of establishing an active locus. PMID:10562551

Activity of human papillomavirus type 16 P97 promoter in immortal and tumorigenic human oral keratinocytes.

PubMed

Kook, J K; Kim, J H; Min, B M

1998-10-01

We previously immortalized normal human oral keratinocytes (NHOK) by transfection with cloned human papillomavirus type 16 (HPV-16) genome and converted these immortalized cells to tumorigenic cells with chemical carcinogens. Since the tumorigenic cells expressed higher level of HPV-16 E6/E7 transcripts, we predicted that enhanced E6/E7 expression was induced by mutations at the long control region (LCR) of the viral genome integrated into cellular chromosome. To test this possibility, we sequenced the entire HPV-16 LCR from immortalized and tumorigenic cells, but no difference in the sequences in all of the tested cells was observed. However, it is possible that such differences in the expression of E6/E7 could have originated from different activities of cellular transcription factors in the different cells. To examine this prospect, we subcloned entire LCR into a reporter gene and determined the promoter activity of LCR in immortalized and tumorigenic cells. We found that the LCR promoter activity was significantly higher in tumorigenic cells when comparing to immortalized cells. We also observed that at least 477 nucleotides upstream of E6 open reading frame are needed for the maximum LCR promoter activity in tumorigenic cells.

Molecular phylogenetics reveals convergent evolution in lower Congo River spiny eels.

PubMed

Alter, S Elizabeth; Brown, Bianca; Stiassny, Melanie L J

2015-10-15

The lower Congo River (LCR) is a region of exceptional species diversity and endemism in the Congo basin, including numerous species of spiny eels (genus Mastacembelus). Four of these exhibit distinctive phenotypes characterized by greatly reduced optic globes deeply embedded into the head (cryptophthalmia) and reduced (or absent) melanin pigmentation, among other characteristics. A strikingly similar cryptophthalmic phenotype is also found in members of a number of unrelated fish families, strongly suggesting the possibility of convergent evolution. However, little is known about the evolutionary processes that shaped diversification in LCR Mastacembelus, their biogeographic origins, or when colonization of the LCR occurred. We sequenced mitochondrial and nuclear genes from Mastacembelus species collected in the lower Congo River, and compared them with other African species and Asian representatives as outgroups. We analyzed the sequence data using Maximum Likelihood and Bayesian phylogenetic inference. Bayesian and Maximum Likelihood phylogenetic analyses, and Bayesian coalescent methods for species tree reconstruction, reveal that endemic LCR spiny eels derive from two independent origins, clearly demonstrating convergent evolution of the cryptophthalmic phenotype. Mastacembelus crassus, M. aviceps, and M. simbi form a clade, allied to species found in southern, eastern and central Africa. Unexpectedly, M. brichardi and brachyrhinus fall within a clade otherwise endemic to Lake Tanganikya (LT) ca. 1500 km east of the LCR. Divergence dating suggests the ages of these two clades of LCR endemics differ markedly. The age of the crassus group is estimated at ~4 Myr while colonization of the LCR by the brichardi-brachyrhinus progenitor was considerably more recent, dated at ~0.5 Myr. The phylogenetic framework of spiny eels presented here, the first to include LCR species, demonstrates that cryptophthalmia and associated traits evolved at least twice in Mastacembelus: once in M. brichardi and at least once in the M. crassus clade. Timing of diversification is broadly consistent with the onset of modern high-energy flow conditions in the LCR and with previous studies of endemic cichlids. The close genetic relationship between M. brichardi and M. brachyrhinus is particularly notable given the extreme difference in phenotype between these species, and additional work is needed to better understand the evolutionary history of diversification in this clade. The findings presented here demonstrate strong, multi-trait convergence in LCR spiny eels, suggesting that extreme selective pressures have shaped numerous phenotypic attributes of the endemic species of this region.

Surgery results in exaggerated and persistent cognitive decline in a rat model of the Metabolic Syndrome.

PubMed

Feng, Xiaomei; Degos, Vincent; Koch, Lauren G; Britton, Steven L; Zhu, Yinggang; Vacas, Susana; Terrando, Niccolò; Nelson, Jeffrey; Su, Xiao; Maze, Mervyn

2013-05-01

Postoperative cognitive decline can be reproduced in animal models. In a well-validated rat model of the Metabolic Syndrome, we sought to investigate whether surgery induced a more severe and persistent form of cognitive decline similar to that noted in preliminary clinical studies. In rats that had been selectively bred for low and high exercise endurance, the low capacity runners (LCR) exhibited features of Metabolic Syndrome (obesity, dyslipidemia, insulin resistance, and hypertension). Tibial fracture surgery was performed under isoflurane anesthesia in LCR and high capacity runner (HCR) rats and cognitive function was assessed postoperatively in a trace-fear conditioning paradigm and Morris Water Maze; non-operated rats were exposed to anesthesia and analgesia (sham). Group sizes were n = 6. On postoperative D7, LCR rats had shorter freezing times than postoperative HCR rats. Five months postoperatively, LCR rats had a flatter learning trajectory and took longer to locate the submerged platform than postoperative HCR rats; dwell-time in the target quadrant in a probe trial was shorter in the postoperative LCR compared to HCR rats. LCR and HCR sham rats did not differ in any test. Postoperatively, LCR rats diverged from HCR rats exhibiting a greater decline in memory, acutely, with persistent learning and memory decline, remotely; this could not be attributed to changes in locomotor or swimming performance. This Metabolic Syndrome animal model of surgery-induced cognitive decline corroborates, with high fidelity, preliminary findings of postoperative cognitive dysfunction in Metabolic Syndrome patients.

CpG methylation in human papillomavirus (HPV) type 31 long control region (LCR) in cervical infections associated with cytological abnormalities.

PubMed

László, Brigitta; Ferenczi, Annamária; Madar, László; Gyöngyösi, Eszter; Szalmás, Anita; Szakács, Levente; Veress, György; Kónya, József

2016-08-01

The mechanisms that regulate papillomavirus gene expression include DNA methylation. The transcription of papillomavirus oncogenes E6 and E7 is controlled by certain regulatory elements in the LCR, which include binding sites for the E2 protein, a viral regulator of oncogene expression. In HPV-31-infected exfoliated cervical cells, the CpG methylation of the entire LCR was determined by next-generation sequencing after bisulfite modification. Six of the 22 cases had methylated CpG sites in the HPV-31 LCR, including position 7479 and/or 7485, at the promoter distal E2 binding site, thus suggesting a potential regulatory mechanism for papillomavirus transcription.

Oral vaccination with different antigens from Yersinia pestis KIM delivered by live attenuated Salmonella typhimurium elicits a protective immune response against plague.

PubMed

Branger, Christine G; Fetherston, Jacqueline D; Perry, Robert D; Curtiss, Roy

2007-01-01

The use of live recombinant Salmonella attenuated vaccine (RASV) encoding Yersinia proteins is a promising new approach for the vaccination against Yersinia pestis. We have tested the efficacy of 2 proteins, Psn and a portion of LcrV in protecting mice against virulent Yersinia pestis challenge. To remove the immunosuppressive properties of LcrV protein, the lcrV gene, without the TLR2 receptor sequence, was cloned into a beta-lactamase secretion vector. Immunizations were performed with RSAV expressing LcrV or Psn. Challenge with a virulent Y. pestis strain was performed 4 weeks after the last immunization. Our results show that the truncated LcrV protein delivered by RASV is sufficient to afford a full protective immune response in a mouse model of bubonic plague and the Psn protein afforded partial protection in a non-optimized system. This finding should facilitate the design and development of a new generation of vaccines against Y. pestis.

The search for early markers of plague: evidence for accumulation of soluble Yersinia pestis LcrV in bubonic and pneumonic mouse models of disease.

PubMed

Flashner, Yehuda; Fisher, Morly; Tidhar, Avital; Mechaly, Adva; Gur, David; Halperin, Gideon; Zahavy, Eran; Mamroud, Emanuelle; Cohen, Sara

2010-07-01

Markers of the early stages of plague, a rapidly progressing deadly disease, are crucial for enabling the onset of an effective treatment. Here, we show that V-antigen protein (LcrV) is accumulated in the serum of Yersinia pestis-infected mice before bacterial colonization of the spleen and dissemination to blood, in a model of bubonic plague. LcrV accumulation is detected earlier than that of F1 capsular antigen, an established marker of disease. In a mouse model of pneumonic plague, LcrV can be determined in the bronchoalveolar lavage fluid somewhat later than F1, but before dissemination of Y. pestis to the blood. Thus, determination of soluble LcrV is suggested as a potential useful tool for monitoring disease progression in both bubonic and pneumonic plague. Moreover, it may be of particular advantage in cases of infections with F1 nonproducing strains.

Evaluation of sub-microsecond recovery resonators for In Vivo Electron Paramagnetic Resonance Imaging

PubMed Central

F, Hyodo; S, Subramanian; N, Devasahayam; R, Murugesan; K, Matsumoto; JB, Mitchell; MC, Krishna

2008-01-01

Time-domain (TD) electron paramagnetic resonance (EPR) imaging at 300 MHz for in vivo applications requires resonators with recovery times less than 1 microsecond after pulsed excitation to reliably capture the rapidly decaying free induction decay (FID). In this study, we tested the suitability of the Litz foil coil resonator (LCR), commonly used in MRI, for in vivo EPR/EPRI applications in the TD mode and compared with parallel coil resonator (PCR). In TD mode, the sensitivity of LCR was lower than that of the PCR. However, in continuous wave (CW) mode, the LCR showed better sensitivity. The RF homogeneity was similar in both the resonators. The axis of the RF magnetic field is transverse to the cylindrical axis of the LCR, making the resonator and the magnet co-axial. Therefore, the loading of animals, and placing of the anesthesia nose cone and temperature monitors was more convenient in the LCR compared to the PCR whose axis is perpendicular to the magnet axis. PMID:18042414

Insulators to improve expression of a 3(')IgH LCR-driven reporter gene in transgenic mouse models.

PubMed

Guglielmi, Laurence; Le Bert, Marc; Truffinet, Véronique; Cogné, Michel; Denizot, Yves

2003-08-01

A locus control region (LCR) containing four transcriptional enhancers lies downstream of the IgH chain locus. We studied transgenes carrying a 3(')IgH LCR-driven GFP reporter gene for expression and B cell differentiation stage specificity. We also compared transgenes that were or were not flanked by two copies of the beta-globin HS4 insulator, an element defined by its ability to protect transgenes from the influences of surrounding genes at the insertion site. Results indicate that insulators are instrumental in sustaining GFP expression in GFP-3(')LCR transgenic mice when they were included. Flow cytometry experiments reported a strictly B cell specific GFP expression from pre-B cells in bone marrow to mature B cells in spleen. Despite addition of 5(')HS4 insulators to the GFP-3(')LCR construct, complete transgene silencing occurred in some transgenic lines and was systematically observed in ageing animals from all lines.

Platelet parameters (PLT, MPV, P-LCR) in patients with schizophrenia, unipolar depression and bipolar disorder.

PubMed

Wysokiński, Adam; Szczepocka, Ewa

2016-03-30

There are no studies comparing platelet parameters platelet parameters (platelet count (PLT), mean platelet volume (MPV) and platelet large cell ratio (P-LCR)) between patients with schizophrenia, bipolar disorder and unipolar depression. Therefore, the aim of this study was to determine and compare differences in PLT, MPV and P-LCR in patients with schizophrenia, unipolar depression and bipolar disorder. This was a retrospective, cross-sectional, naturalistic study of 2377 patients (schizophrenia n=1243; unipolar depression n=791; bipolar disorder n=343, including bipolar depression n=259 and mania n=84). There were significant differences for PLT, MPV and P-LCR values between study groups. A significant percentage of patients with bipolar disorder had abnormal (too low or too high) number of platelets. Negative correlation between PLT and age was found in all study groups and positive correlation between age and MPV and P-LCR was found in patients with schizophrenia. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Vaginal impact of the oral administration of total freeze-dried culture of LCR 35 in healthy women.

PubMed

Bohbot, J M; Cardot, J M

2012-01-01

The use of probiotics in the prevention or treatment of some vaginal infections has been the subject of numerous studies. To assess the presence of Lactobacillus casei rhamnosus (LCR35) in the vagina after an oral administration, an open randomised pilot study was conducted on 20 healthy women of child-bearing age. Materials and Methods. 2 groups of 10 women were given a 28-day oral course, that is, at least 108 CFU/day (group 1) or 2 × 108 CFU/day (group 2) of LCR35. Nugent score and vaginal screening for LCR35 were undertaken before and after 28 days of treatment. Results. The mean Nugent score decreased in group 1 (-0,2) as well as in group 2 (-0,3). 10% of women in group 1 versus 40% of women in group 2 were carrying LCR35 at the end of the trial. Conclusion. LCR35, at the minimal dose of 2 × 108 CFU/day, can return the Nugent score to normal in healthy women of child-bearing age, by means of a well-tolerated vaginal temporary presence. Phase III clinical trials will specify the preventive or curative impact of this orally administered strain on a range of vaginal disorders such as bacterial vaginosis or vulvovaginal candidiasis.

A multicenter phase II study of salvage photodynamic therapy using talaporfin sodium (ME2906) and a diode laser (PNL6405EPG) for local failure after chemoradiotherapy or radiotherapy for esophageal cancer

PubMed Central

Yano, Tomonori; Kasai, Hiroi; Horimatsu, Takahiro; Yoshimura, Kenichi; Teramukai, Satoshi; Morita, Satoshi; Tada, Harue; Yamamoto, Yoshinobu; Kataoka, Hiromi; Kakushima, Naomi; Ishihara, Ryu; Isomoto, Hajime; Muto, Manabu

2017-01-01

Photodynamic therapy (PDT) showed promising efficacy for local failure after chemoradiotherapy (CRT) for esophageal cancer. However, PDT required long sun shade period. This study aimed to evaluate the safety and efficacy of PDT using second generation photosensitizer, talaporfin sodium for local failure after CRT. This was the multi-institutional non-randomized phase II study. Patients with histologically proven local failure limited within the muscularis propria after 50Gy or more radiotherapy (RT) for esophageal cancer were eligible. We set the primary endpoint as local complete response (L-CR) per patients. And, secondary endpoints were confirmed L-CR, local progression free survival (L-PFS), progression free survival (PFS), overall survival (OS), L-CR per lesions (Lesion L-CR), and confirmed Lesion L-CR. The PDT procedure commenced with intravenous administration of a 40 mg/m2 dose of talaporfin sodium followed by diode laser irradiation at a 664 nm wavelength. 26 eligible patients were enrolled and all were treated with PDT. Twenty three patients with 25 lesions were assessed L-CR after PDT; the L-CR rate per patients was 88.5% (95% CI: 69.8%-97.6%). No skin phototoxicity was observed, and no grade 3 or worse non-hematological toxicities related to PDT were observed. PDT using talaporfin sodium and a diode laser is a safe and curative salvage treatment for local failure after CRT or RT for patients with esophageal cancer. PMID:28212527

Sequential changes in chromatin structure during transcriptional activation in the beta globin LCR and its target gene.

PubMed

Kim, Kihoon; Kim, AeRi

2010-09-01

Chromatin structure is modulated during transcriptional activation. The changes include the association of transcriptional activators, formation of hypersensitive sites and covalent modifications of histones. To understand the order of the various changes accompanying transcriptional activation, we analyzed the mouse beta globin gene, which is transcriptionally inducible in erythroid MEL cells over a time course of HMBA treatment. Transcription of the globin genes requires the locus control region (LCR) consisting of several hypersensitive sites (HSs). Erythroid specific transcriptional activators such as NF-E2, GATA-1, TAL1 and EKLF were associated with the LCR in the uninduced state before transcriptional activation. The HSs of the LCR were formed in this state as revealed by high sensitivity to DNase I and MNase attack. However the binding of transcriptional activators and the depletion of histones were observed in the promoter of the beta globin gene only after transcriptional activation. In addition, various covalent histone modifications were sequentially detected in lysine residues of histone H3 during the activation. Acetylation of K9, K36 and K27 was notable in both LCR HSs and gene after induction but before transcriptional initiation. Inactive histone marks such as K9me2, K36me2 and K27me2 were removed coincident with transcriptional initiation in the gene region. Taken together, these results indicate that LCR has a substantially active structure in the uninduced state while transcriptional activation serially adds active marks, including histone modifications, and removes inactive marks in the target gene of the LCR. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

Generation of human auditory steady-state responses (SSRs). II: Addition of responses to individual stimuli.

PubMed

Santarelli, R; Maurizi, M; Conti, G; Ottaviani, F; Paludetti, G; Pettorossi, V E

1995-03-01

In order to investigate the generation of the 40 Hz steady-state response (SSR), auditory potentials evoked by clicks were recorded in 16 healthy subjects in two stimulating conditions. Firstly, repetition rates of 7.9 and 40 Hz were used to obtain individual middle latency responses (MLRs) and 40 Hz-SSRs, respectively. In the second condition, eight click trains were presented at a 40 Hz repetition rate and an inter-train interval of 126 ms. We extracted from the whole train response: (1) the response-segment taking place after the last click of the train (last click response, LCR), (2) a modified LCR (mLCR) obtained by clearing the LCR from the amplitude enhancement due to the overlapping of the responses to the clicks preceding the last within the stimulus train. In comparison to MLRs, the most relevant feature of the evoked activity following the last click of the train (LCRs, mLCRs) was the appearance in the 50-110 ms latency range of one (in 11 subjects) or two (in 2 subjects) additional positive-negative deflections having the same periodicity as that of MLR waves. The grand average (GA) of the 40 Hz-SSRs was compared with three predictions synthesized by superimposing: (1) the GA of MLRs, (2) the GA of LCRs, (3) the GA of mLCRs. Both the MLR and mLCR predictions reproduced the recorded signal in amplitude while the LCR prediction amplitude resulted almost twice that of the 40 Hz-SSR. With regard to the phase, the MLR, LCR and mLCR closely predicted the recorded signal. Our findings confirm the effectiveness of the linear addition mechanism in the generation of the 40 Hz-SSR. However the responses to individual stimuli within the 40 Hz-SSR differ from MLRs because of additional periodic activity. These results suggest that phenomena related to the resonant frequency of the activated system may play a role in the mechanisms which interact to generate the 40 Hz-SSR.

Beta-globin LCR and intron elements cooperate and direct spatial reorganization for gene therapy.

PubMed

Buzina, Alla; Lo, Mandy Y M; Moffett, Angela; Hotta, Akitsu; Fussner, Eden; Bharadwaj, Rikki R; Pasceri, Peter; Garcia-Martinez, J Victor; Bazett-Jones, David P; Ellis, James

2008-04-11

The Locus Control Region (LCR) requires intronic elements within beta-globin transgenes to direct high level expression at all ectopic integration sites. However, these essential intronic elements cannot be transmitted through retrovirus vectors and their deletion may compromise the therapeutic potential for gene therapy. Here, we systematically regenerate functional beta-globin intron 2 elements that rescue LCR activity directed by 5'HS3. Evaluation in transgenic mice demonstrates that an Oct-1 binding site and an enhancer in the intron cooperate to increase expression levels from LCR globin transgenes. Replacement of the intronic AT-rich region with the Igmu 3'MAR rescues LCR activity in single copy transgenic mice. Importantly, a combination of the Oct-1 site, Igmu 3'MAR and intronic enhancer in the BGT158 cassette directs more consistent levels of expression in transgenic mice. By introducing intron-modified transgenes into the same genomic integration site in erythroid cells, we show that BGT158 has the greatest transcriptional induction. 3D DNA FISH establishes that induction stimulates this small 5'HS3 containing transgene and the endogenous locus to spatially reorganize towards more central locations in erythroid nuclei. Electron Spectroscopic Imaging (ESI) of chromatin fibers demonstrates that ultrastructural heterochromatin is primarily perinuclear and does not reorganize. Finally, we transmit intron-modified globin transgenes through insulated self-inactivating (SIN) lentivirus vectors into erythroid cells. We show efficient transfer and robust mRNA and protein expression by the BGT158 vector, and virus titer improvements mediated by the modified intron 2 in the presence of an LCR cassette composed of 5'HS2-4. Our results have important implications for the mechanism of LCR activity at ectopic integration sites. The modified transgenes are the first to transfer intronic elements that potentiate LCR activity and are designed to facilitate correction of hemoglobinopathies using single copy vectors.

Serotonin in the solitary tract nucleus shortens the laryngeal chemoreflex in anaesthetized neonatal rats.

PubMed

Donnelly, William T; Bartlett, Donald; Leiter, J C

2016-07-01

What is the central question of this study? Failure to terminate apnoea and arouse is likely to contribute to sudden infant death syndrome (SIDS). Serotonin is deficient in the brainstems of babies who died of SIDS. Therefore, we tested the hypothesis that serotonin in the nucleus of the solitary tract (NTS) would shorten reflex apnoea. What is the main finding and its importance? Serotonin microinjected into the NTS shortened the apnoea and respiratory inhibition associated with the laryngeal chemoreflex. Moreover, this effect was achieved through a 5-HT3 receptor. This is a new insight that is likely to be relevant to the pathogenesis of SIDS. The laryngeal chemoreflex (LCR), an airway-protective reflex that causes apnoea and bradycardia, has long been suspected as an initiating event in the sudden infant death syndrome. Serotonin (5-HT) and 5-HT receptors may be deficient in the brainstems of babies who die of sudden infant death syndrome, and 5-HT seems to be important in terminating apnoeas directly or in causing arousals or as part of the process of autoresuscitation. We hypothesized that 5-HT in the brainstem would limit the duration of the LCR. We studied anaesthetized rat pups between 7 and 21 days of age and made microinjections into the cisterna magna or into the nucleus of the solitary tract (NTS). Focal, bilateral microinjections of 5-HT into the caudal NTS significantly shortened the LCR. The 5-HT1a receptor antagonist, WAY 100635, did not affect the LCR consistently, nor did a 5-HT2 receptor antagonist, ketanserin, alter the duration of the LCR. The 5-HT3 specific agonist, 1-(3-chlorophenyl)-biguanide, microinjected bilaterally into the caudal NTS significantly shortened the LCR. Thus, endogenous 5-HT released within the NTS may curtail the respiratory depression that is part of the LCR, and serotonergic shortening of the LCR may be attributed to activation of 5-HT3 receptors within the NTS. 5-HT3 receptors are expressed presynaptically on C fibre afferents of the superior laryngeal nerve, and serotonergic shortening of the LCR may be mediated presynaptically by enhanced activation of inhibitory interneurons within the NTS. © 2016 The Authors. Experimental Physiology © 2016 The Physiological Society.

Concerted Actions of a Thermo-labile Regulator and a Unique Intergenic RNA Thermosensor Control Yersinia Virulence

PubMed Central

Kortmann, Jens; Seekircher, Stephanie; Heroven, Ann Kathrin; Berger, Evelin; Pisano, Fabio; Thiermann, Tanja; Wolf-Watz, Hans; Narberhaus, Franz; Dersch, Petra

2012-01-01

Expression of all Yersinia pathogenicity factors encoded on the virulence plasmid, including the yop effector and the ysc type III secretion genes, is controlled by the transcriptional activator LcrF in response to temperature. Here, we show that a protein- and RNA-dependent hierarchy of thermosensors induce LcrF synthesis at body temperature. Thermally regulated transcription of lcrF is modest and mediated by the thermo-sensitive modulator YmoA, which represses transcription from a single promoter located far upstream of the yscW-lcrF operon at moderate temperatures. The transcriptional response is complemented by a second layer of temperature-control induced by a unique cis-acting RNA element located within the intergenic region of the yscW-lcrF transcript. Structure probing demonstrated that this region forms a secondary structure composed of two stemloops at 25°C. The second hairpin sequesters the lcrF ribosomal binding site by a stretch of four uracils. Opening of this structure was favored at 37°C and permitted ribosome binding at host body temperature. Our study further provides experimental evidence for the biological relevance of an RNA thermometer in an animal model. Following oral infections in mice, we found that two different Y. pseudotuberculosis patient isolates expressing a stabilized thermometer variant were strongly reduced in their ability to disseminate into the Peyer's patches, liver and spleen and have fully lost their lethality. Intriguingly, Yersinia strains with a destabilized version of the thermosensor were attenuated or exhibited a similar, but not a higher mortality. This illustrates that the RNA thermometer is the decisive control element providing just the appropriate amounts of LcrF protein for optimal infection efficiency. PMID:22359501

L-Carnitine halts apoptosis and myelosuppression induced by carboplatin in rat bone marrow cell cultures (BMC).

PubMed

Abd-Allah, Adel R A; Al-Majed, Abdulhakeem A; Al-Yahya, Abdulaziz A; Fouda, Soliman I; Al-Shabana, Othman A

2005-07-01

Carboplatin (CP), a second generation platinum compound, is effective against various types of cancers, producing less nephrotoxicity and ototoxicity but more myelotoxicity than cisplatinum. CP-myelosuppression is the rate-limiting step of its clinical use. Prevention of CP-myelosuppression is a major target in the field of chemotherapy. Therefore, the present study investigates the use of L-carnitine (LCR)-an antioxidant, cardioprotective, neuroprotective, and immunostimulant nontoxic natural compound-to protect against CP-induced myelosuppression. The viability of BMC was studied using a trypan blue exclusion technique following incubation with CP and/or LCR as a function of time and concentration. Apoptosis was tested for by detecting the amount of DNA fragmentation and the visualization of DNA ladders upon gel electrophoresis. Bone marrow progenitor cell function was examined by colony forming unit assay. Cellular contents of glutathione (GSH) and malondialdehyde (MDA) were also estimated. Results revealed that LC50 of CP is 4.7 mM and the highest safe concentration of LCR is 5 mM. Co-exposure of LCR+CP rescued BMC viability by 37% compared to the CP-treated cultures. The LCR halts CP-induced apoptosis and it significantly improves the function of the bone marrow progenitors by increasing the number of colony-forming units as a response to granulocyte/macrophage colony stimulating factors. Finally, LCR restores CP-induced GSH depletion and prevents MDA elevation in BMC. In summary, the results suggest that LCR is able to protect against CP-induced myelosuppression, which suggests its use as an adjuvant therapy. This finding merits further investigation into the mechanism(s) of such protection as well as its interaction with CP antitumor activity.

Sex-Specific Consequences of Neonatal Stress on Cardio-Respiratory Inhibition Following Laryngeal Stimulation in Rat Pups

PubMed Central

Baldy, Cécile; Chamberland, Simon

2017-01-01

Abstract The presence of liquid near the larynx of immature mammals triggers prolonged apneas with significant O2 desaturations and bradycardias. When excessive, this reflex (the laryngeal chemoreflex; LCR) can be fatal. Our understanding of the origins of abnormal LCR are limited; however, perinatal stress and male sex are risk factors for cardio-respiratory failure in infants. Because exposure to stress during early life has deleterious and sex-specific consequences on brain development it is plausible that respiratory reflexes are vulnerable to neuroendocrine dysfunction. To address this issue, we tested the hypothesis that neonatal maternal separation (NMS) is sufficient to exacerbate LCR-induced cardio-respiratory inhibition in anesthetized rat pups. Stressed pups were separated from their mother 3 h/d from postnatal days 3 to 12. At P14–P15, pups were instrumented to monitor breathing, O2 saturation (Spo2), and heart rate. The LCR was activated by water injections near the larynx (10 µl). LCR-induced apneas were longer in stressed pups than controls; O2 desaturations and bradycardias were more profound, especially in males. NMS increased the frequency and amplitude of spontaneous EPSCs (sEPSCs) in the dorsal motor nucleus of the vagus (DMNV) of males but not females. The positive relationship between corticosterone and testosterone observed in stressed pups (males only) suggests that disruption of neuroendocrine function by stress is key to sex-based differences in abnormal LCR. Because testosterone application onto medullary slices augments EPSC amplitude only in males, we propose that testosterone-mediated enhancement of synaptic connectivity within the DMNV contributes to the male bias in cardio-respiratory inhibition following LCR activation in stressed pups. PMID:29308430

Effect of variable annual precipitation and nutrient input on nitrogen and phosphorus transport from two Midwestern agricultural watersheds

USGS Publications Warehouse

Kalkhoff, Stephen J.; Hubbard, Laura E.; Tomer, Mark D.; James, D.E.

2016-01-01

Precipitation patterns and nutrient inputs affect transport of nitrate (NO3-N) and phosphorus (TP) from Midwest watersheds. Nutrient concentrations and yields from two subsurface-drained watersheds, the Little Cobb River (LCR) in southern Minnesota and the South Fork Iowa River (SFIR) in northern Iowa, were evaluated during 1996–2007 to document relative differences in timings and amounts of nutrients transported. Both watersheds are located in the prairie pothole region, but the SFIR exhibits a longer growing season and more livestock production. The SFIR yielded significantly more NO3-N than the LCR watershed (31.2 versus 21.3 kg NO3-N ha− 1 y− 1). The SFIR watershed also yielded more TP than the LCR watershed (1.13 versus 0.51 kg TP ha− 1 yr− 1), despite greater TP concentrations in the LCR. About 65% of NO3-N and 50% of TP loads were transported during April–June, and < 20% of the annual loads were transported later in the growing season from July–September. Monthly NO3-N and TP loads peaked in April from the LCR but peaked in June from the SFIR; this difference was attributed to greater snowmelt runoff in the LCR. The annual NO3-N yield increased with increasing annual runoff at a similar rate in both watersheds, but the LCR watershed yielded less annual NO3-N than the SFIR for a similar annual runoff. These two watersheds are within 150 km of one another and have similar dominant agricultural systems, but differences in climate and cropping inputs affected amounts and timing of nutrient transport.

Exaggerated Acute Lung Injury and Impaired Antibacterial Defenses During Staphylococcus aureus Infection in Rats with the Metabolic Syndrome

PubMed Central

Feng, Xiaomei; Maze, Mervyn; Koch, Lauren G.; Britton, Steven L.; Hellman, Judith

2015-01-01

Rats with Metabolic Syndrome (MetaS) have a dysregulated immune response to the aseptic trauma of surgery. We hypothesized that rats with MetaS would have dysregulated inflammation, increased lung injury, and less effective antibacterial defenses during Staphylococcus (S.) aureus sepsis as compared to rats without MetaS. Low capacity runner (LCR; a model of MetaS) and high capacity runner (HCR) rats were challenged intravenously with S. aureus bacteria. After 48 h, inflammatory mediators and bacteria were quantified in the blood, bronchoalveolar lavage fluid (BALF), and lung homogenates. Lungs were analyzed histologically. BALF protein and lung wet-dry ratios were quantified to assess for vascular leak. Endpoints were compared in infected LCR vs HCR rats. LCR rats had higher blood and lung S. aureus counts, as well as higher levels of IL-6 in plasma, lungs and BALF, MIP-2 in plasma and lung, and IL-17A in lungs. Conversely, LCR rats had lower levels of IL-10 in plasma and lungs. Although lactate levels, and liver and renal function tests were similar between groups, LCR rats had higher BALF protein and lung wet-dry ratios, and more pronounced acute lung injury histologically. During S. aureus bacteremia, as compared with HCR rats, LCR (MetaS) rats have heightened pro-inflammatory responses, accompanied by increased acute lung injury and vascular leak. Notably, despite an augmented pro-inflammatory phenotype, LCR rats have higher bacterial levels in their blood and lungs. The MetaS state may exacerbate lung injury and vascular leak by attenuating the inflammation-resolving response, and by weakening antimicrobial defenses. PMID:25978669

Low intrinsic exercise capacity in rats predisposes to age-dependent cardiac remodeling independent of macrovascular function

PubMed Central

Ritchie, Rebecca H.; Leo, Chen Huei; Qin, Chengxue; Stephenson, Erin J.; Bowden, Marissa A.; Buxton, Keith D.; Lessard, Sarah J.; Rivas, Donato A.; Koch, Lauren G.; Britton, Steven L.; Woodman, Owen L.

2013-01-01

Rats selectively bred for low (LCR) or high (HCR) intrinsic running capacity simultaneously present with contrasting risk factors for cardiovascular and metabolic disease. However, the impact of these phenotypes on left ventricular (LV) morphology and microvascular function, and their progression with aging, remains unresolved. We tested the hypothesis that the LCR phenotype induces progressive age-dependent LV remodeling and impairments in microvascular function, glucose utilization, and β-adrenergic responsiveness, compared with HCR. Hearts and vessels isolated from female LCR (n = 22) or HCR (n = 26) were studied at 12 and 35 wk. Nonselected N:NIH founder rats (11 wk) were also investigated (n = 12). LCR had impaired glucose tolerance and elevated plasma insulin (but not glucose) and body-mass at 12 wk compared with HCR, with early LV remodeling. By 35 wk, LV prohypertrophic and glucose transporter GLUT4 gene expression were up- and downregulated, respectively. No differences in LV β-adrenoceptor expression or cAMP content between phenotypes were observed. Macrovascular endothelial function was predominantly nitric oxide (NO)-mediated in both phenotypes and remained intact in LCR for both age-groups. In contrast, mesenteric arteries microvascular endothelial function, which was impaired in LCR rats regardless of age. At 35 wk, endothelial-derived hyperpolarizing factor-mediated relaxation was impaired whereas the NO contribution to relaxation is intact. Furthermore, there was reduced β2-adrenoceptor responsiveness in both aorta and mesenteric LCR arteries. In conclusion, diminished intrinsic exercise capacity impairs systemic glucose tolerance and is accompanied by progressive development of LV remodeling. Impaired microvascular perfusion is a likely contributing factor to the cardiac phenotype. PMID:23262135

Dysfunction of Inflammation-Resolving Pathways Is Associated with Exaggerated Postoperative Cognitive Decline in a Rat Model of the Metabolic Syndrome

PubMed Central

Su, Xiao; Feng, Xiaomei; Terrando, Niccolo; Yan, Yan; Chawla, Ajay; Koch, Lauren G; Britton, Steven L; Matthay, Michael A; Maze, Mervyn

2012-01-01

The cholinergic antiinflammatory pathway (CAP), which terminates in the spleen, attenuates postoperative cognitive decline (PCD) in rodents. Surgical patients with metabolic syndrome exhibit exaggerated and persistent PCD that is reproduced in postoperative rats selectively bred for easy fatigability and that contain all features of metabolic syndrome (low-capacity runners [LCRs]). We compared the CAP and lipoxin A4 (LXA4), another inflammation-resolving pathway in LCR, with its counterpart high-capacity runner (HCR) rats. Isoflurane-anesthetized LCR and HCR rats either underwent aseptic trauma involving tibial fracture (surgery) or not (sham). At postoperative d 3 (POD3), compared with HCR, LCR rats exhibited significantly exaggerated PCD (trace fear conditioning freezing time 43% versus 57%). Separate cohorts were killed at POD3 to collect plasma for LXA4 and to isolate splenic mononuclear cells (MNCs) to analyze CAP signaling, regulatory T cells (Tregs) and M2 macrophages (M2 Mφ). Under lipopolysaccharide (LPS) stimulation, tumor necrosis factor (TNF)-α produced by splenic MNCs was 117% higher in LCR sham and 52% higher in LCR surgery compared with HCR sham and surgery rats; LPS-stimulated TNF-α production could not be inhibited by an α7 nicotinic acetylcholine receptor agonist, whereas inhibition by the β2 adrenergic agonist, salmeterol, was significantly less (−35%) than that obtained in HCR rats. Compared to HCR, sham and surgery LCR rats had reduced β2 adrenergic receptor–expressing T lymphocytes (59%, 44%), Tregs (47%, 54%) and M2 Mφ (45%, 39%); surgical LCR rats’ hippocampal M2 Mφ was 66% reduced, and plasma LXA4 was decreased by 120%. Rats with the metabolic syndrome have ineffective inflammation-resolving mechanisms that represent plausible reasons for the exaggerated and persistent PCD. PMID:23296426

Concerted actions of a thermo-labile regulator and a unique intergenic RNA thermosensor control Yersinia virulence.

PubMed

Böhme, Katja; Steinmann, Rebekka; Kortmann, Jens; Seekircher, Stephanie; Heroven, Ann Kathrin; Berger, Evelin; Pisano, Fabio; Thiermann, Tanja; Wolf-Watz, Hans; Narberhaus, Franz; Dersch, Petra

2012-02-01

Expression of all Yersinia pathogenicity factors encoded on the virulence plasmid, including the yop effector and the ysc type III secretion genes, is controlled by the transcriptional activator LcrF in response to temperature. Here, we show that a protein- and RNA-dependent hierarchy of thermosensors induce LcrF synthesis at body temperature. Thermally regulated transcription of lcrF is modest and mediated by the thermo-sensitive modulator YmoA, which represses transcription from a single promoter located far upstream of the yscW-lcrF operon at moderate temperatures. The transcriptional response is complemented by a second layer of temperature-control induced by a unique cis-acting RNA element located within the intergenic region of the yscW-lcrF transcript. Structure probing demonstrated that this region forms a secondary structure composed of two stemloops at 25°C. The second hairpin sequesters the lcrF ribosomal binding site by a stretch of four uracils. Opening of this structure was favored at 37°C and permitted ribosome binding at host body temperature. Our study further provides experimental evidence for the biological relevance of an RNA thermometer in an animal model. Following oral infections in mice, we found that two different Y. pseudotuberculosis patient isolates expressing a stabilized thermometer variant were strongly reduced in their ability to disseminate into the Peyer's patches, liver and spleen and have fully lost their lethality. Intriguingly, Yersinia strains with a destabilized version of the thermosensor were attenuated or exhibited a similar, but not a higher mortality. This illustrates that the RNA thermometer is the decisive control element providing just the appropriate amounts of LcrF protein for optimal infection efficiency.

Vaginal Impact of the Oral Administration of Total Freeze-Dried Culture of LCR 35 in Healthy Women

PubMed Central

Bohbot, J. M.; Cardot, J. M.

2012-01-01

The use of probiotics in the prevention or treatment of some vaginal infections has been the subject of numerous studies. To assess the presence of Lactobacillus casei rhamnosus (LCR35) in the vagina after an oral administration, an open randomised pilot study was conducted on 20 healthy women of child-bearing age. Materials and Methods. 2 groups of 10 women were given a 28-day oral course, that is, at least 108 CFU/day (group 1) or 2 × 108 CFU/day (group 2) of LCR35. Nugent score and vaginal screening for LCR35 were undertaken before and after 28 days of treatment. Results. The mean Nugent score decreased in group 1 (−0,2) as well as in group 2 (−0,3). 10% of women in group 1 versus 40% of women in group 2 were carrying LCR35 at the end of the trial. Conclusion. LCR35, at the minimal dose of 2 × 108 CFU/day, can return the Nugent score to normal in healthy women of child-bearing age, by means of a well-tolerated vaginal temporary presence. Phase III clinical trials will specify the preventive or curative impact of this orally administered strain on a range of vaginal disorders such as bacterial vaginosis or vulvovaginal candidiasis. PMID:22701297

Brain Activation Patterns at Exhaustion in Rats That Differ in Inherent Exercise Capacity

PubMed Central

Foley, Teresa E.; Brooks, Leah R.; Gilligan, Lori J.; Burghardt, Paul R.; Koch, Lauren G.; Britton, Steven L.; Fleshner, Monika

2012-01-01

In order to further understand the genetic basis for variation in inherent (untrained) exercise capacity, we examined the brains of 32 male rats selectively bred for high or low running capacity (HCR and LCR, respectively). The aim was to characterize the activation patterns of brain regions potentially involved in differences in inherent running capacity between HCR and LCR. Using quantitative in situ hybridization techniques, we measured messenger ribonuclease (mRNA) levels of c-Fos, a marker of neuronal activation, in the brains of HCR and LCR rats after a single bout of acute treadmill running (7.5–15 minutes, 15° slope, 10 m/min) or after treadmill running to exhaustion (15–51 minutes, 15° slope, initial velocity 10 m/min). During verification of trait differences, HCR rats ran six times farther and three times longer prior to exhaustion than LCR rats. Running to exhaustion significantly increased c-Fos mRNA activation of several brain areas in HCR, but LCR failed to show significant elevations of c-Fos mRNA at exhaustion in the majority of areas examined compared to acutely run controls. Results from these studies suggest that there are differences in central c-Fos mRNA expression, and potential brain activation patterns, between HCR and LCR rats during treadmill running to exhaustion and these differences could be involved in the variation in inherent running capacity between lines. PMID:23028992

All of the human beta-type globin genes compete for LCR enhancer activity in embryonic erythroid cells of yeast artificial chromosome transgenic mice.

PubMed

Okamura, Eiichi; Matsuzaki, Hitomi; Campbell, Andrew D; Engel, James Douglas; Fukamizu, Akiyoshi; Tanimoto, Keiji

2009-12-01

In primitive erythroid cells of human beta-globin locus transgenic mice (TgM), the locus control region (LCR)-proximal epsilon- and gamma-globin genes are transcribed, whereas the distal delta- and beta-globin genes are silent. It is generally accepted that the beta-globin gene is competitively suppressed by gamma-globin gene expression at this developmental stage. Previously, however, we observed that epsilon-globin gene expression was severely attenuated when its distance from the LCR was extended, implying that beta-globin gene might also be silenced because of its great distance from the LCR. Here, to clarify the beta-globin gene silencing mechanism, we established TgM lines carrying either gamma- or epsilon- plus gamma-globin promoter deletions, without significantly altering the distance between the beta-globin gene and the LCR. Precocious expression of delta- and beta-globin genes was observed in primitive erythroid cells of mutant, but not wild-type TgM, which was most evident when both the epsilon and gamma promoters were deleted. Thus, we clearly demonstrated that the repression of the delta- and beta-globin genes in primitive erythroid cells is dominated by competitive silencing by the epsilon- and gamma-globin gene promoters, and that epsilon- and the other beta-like globin genes might be activated by two distinct mechanisms by the LCR.

Purification and characterization of Yersinia enterocolitica and Yersinia pestis LcrV-cholera toxin A(2)/B chimeras.

PubMed

Tinker, Juliette K; Davis, Chadwick T; Arlian, Britni M

2010-11-01

Yersinia pestis is a virulent human pathogen and potential biological weapon. Despite a long history of research on this organism, there is no licensed vaccine to protect against pneumonic forms of Y. pestis disease. In the present study, plasmids were constructed to express cholera toxin A(2)/B chimeric molecules containing the LcrV protective antigen from Yersinia enterocolitica and Y. pestis. These chimeras were expressed and purified to high yields from the supernatant of transformed Escherichia coli. Western and GM(1) ELISA assays were used to characterize the composition, receptor-binding and relative stability of the LcrV-CTA(2)/B chimera in comparison to cholera toxin. In addition, we investigated the ability of the Y. pestis LcrV-CTA(2)/B chimera to bind to and internalize into cultured epithelial cells and macrophages by confocal microscopy. These studies indicate that the uptake and trafficking of the LcrV antigen from the chimera is comparable to the trafficking of native toxin. Together these findings report that stable, receptor-binding, non-toxic LcrV-cholera toxin A(2)/B chimeras can be expressed at high levels in E. coli and purified from the supernatant. In addition, the internalization of antigen in vitro reported here supports the development of these molecules as novel mucosal vaccine candidates. Copyright 2010 Elsevier Inc. All rights reserved.

Purification and characterization of Yersinia enterocolitica and Yersinia pestis LcrV–cholera toxin A2/B chimeras

PubMed Central

Davis, Chadwick T.; Arlian, Britni M.

2010-01-01

Yersinia pestis is a virulent human pathogen and potential biological weapon. Despite a long history of research on this organism, there is no licensed vaccine to protect against pneumonic forms of Y. pestis disease. In the present study, plasmids were constructed to express cholera toxin A2/B chimeric molecules containing the LcrV protective antigen from Y. enterocolitica and Y. pestis. These chimeras were expressed and purified to high yields from the supernatant of transformed E. coli. Western and GM1 ELISA assays were used to characterize the composition, receptor-binding and relative stability of the LcrV-CTA2/B chimera in comparison to cholera toxin. In addition, we investigated the ability of the Y. pestis LcrV-CTA2/B chimera to bind to and internalize into cultured epithelial cells and macrophages by confocal microscopy. These studies indicate that the uptake and trafficking of the LcrV antigen from the chimera is comparable to the trafficking of native toxin. Together these findings report that stable, receptor-binding, non-toxic LcrV-cholera toxin A2/B chimeras can be expressed at high levels in E. coli and purified from the supernatant. In addition, the internalization of antigen in vitro reported here supports the development of these molecules as novel mucosal vaccine candidates. PMID:20438844

Fretting fatigue behaviour of Ni-free high-nitrogen stainless steel in a simulated body fluid.

PubMed

Maruyama, Norio; Hiromoto, Sachiko; Akiyama, Eiji; Nakamura, Morihiko

2013-04-01

Fretting fatigue behaviour of Ni-free high-nitrogen steel (HNS) with a yield strength of about 800 MPa, which was prepared by nitrogen gas pressurized electroslag remelting, was studied in air and in phosphate-buffered saline (PBS(-)). For comparison, fretting fatigue behaviour of cold-rolled SUS316L steel (SUS316L(CR)) with similar yield strength was examined. The plain fatigue limit of HNS was slightly lower than that of SUS316L(CR) although the former had a higher tensile strength than the latter. The fretting fatigue limit of HNS was higher than that of SUS316L(CR) both in air and in PBS(-). A decrease in fatigue limit of HNS by fretting was significantly smaller than that of SUS316L(CR) in both environments, indicating that HNS has better fretting fatigue resistance than SUS316L(CR). The decrease in fatigue limit by fretting is discussed taking into account the effect of friction stress due to fretting and the additional influences of wear, tribocorrosion and plastic deformation in the fretted area.

Fretting fatigue behaviour of Ni-free high-nitrogen stainless steel in a simulated body fluid

NASA Astrophysics Data System (ADS)

Maruyama, Norio; Hiromoto, Sachiko; Akiyama, Eiji; Nakamura, Morihiko

2013-04-01

Fretting fatigue behaviour of Ni-free high-nitrogen steel (HNS) with a yield strength of about 800 MPa, which was prepared by nitrogen gas pressurized electroslag remelting, was studied in air and in phosphate-buffered saline (PBS(-)). For comparison, fretting fatigue behaviour of cold-rolled SUS316L steel (SUS316L(CR)) with similar yield strength was examined. The plain fatigue limit of HNS was slightly lower than that of SUS316L(CR) although the former had a higher tensile strength than the latter. The fretting fatigue limit of HNS was higher than that of SUS316L(CR) both in air and in PBS(-). A decrease in fatigue limit of HNS by fretting was significantly smaller than that of SUS316L(CR) in both environments, indicating that HNS has better fretting fatigue resistance than SUS316L(CR). The decrease in fatigue limit by fretting is discussed taking into account the effect of friction stress due to fretting and the additional influences of wear, tribocorrosion and plastic deformation in the fretted area.

The human desmin locus: gene organization and LCR-mediated transcriptional control.

PubMed

Tam, Jennifer L Y; Triantaphyllopoulos, Kostas; Todd, Helen; Raguz, Selina; de Wit, Ton; Morgan, Jennifer E; Partridge, Terence A; Makrinou, Eleni; Grosveld, Frank; Antoniou, Michael

2006-06-01

Locus control regions (LCRs) are defined by their ability to confer reproducible physiological levels of transgene expression in mice and therefore thought to possess the ability to generate dominantly a transcriptionally active chromatin structure. We report the first characterization of a muscle-cell-specific LCR, which is linked to the human desmin gene (DES). The DES LCR consists of five regions of muscle-specific DNase I hypersensitivity (HS) localized between -9 and -18 kb 5' of DES and reproducibly drives full physiological levels of expression in all muscle cell types. The DES LCR DNase I HS regions are highly conserved between humans and other mammals and can potentially bind a broad range of muscle-specific and ubiquitous transcription factors. Bioinformatics and direct molecular analysis show that the DES locus consists of three muscle-specific (DES) or muscle preferentially expressed genes (APEG1 and SPEG, the human orthologue of murine striated-muscle-specific serine/threonine protein kinase, Speg). The DES LCR may therefore regulate expression of SPEG and APEG1 as well as DES.

Oral vaccination with LcrV from Yersinia pestis KIM delivered by live attenuated Salmonella enterica serovar Typhimurium elicits a protective immune response against challenge with Yersinia pseudotuberculosis and Yersinia enterocolitica.

PubMed

Branger, Christine G; Torres-Escobar, Ascención; Sun, Wei; Perry, Robert; Fetherston, Jacqueline; Roland, Kenneth L; Curtiss, Roy

2009-08-27

The use of live recombinant attenuated Salmonella vaccines (RASV) synthesizing Yersinia proteins is a promising approach for controlling infection by Yersinia species. In this study, we constructed attenuated Salmonella strains which synthesize a truncated form of LcrV, LcrV196 and evaluated the immune response and protective efficacy elicited by these strains in mice against two other major species of Yersinia: Yersinia pseudotuberculosis and Yersinia enterocolitica. Surprisingly, we found that the RASV strain alone was sufficient to afford nearly full protection against challenge with Y. pseudotuberculosis, indicating the likelihood that Salmonella produces immunogenic cross-protective antigens. In contrast, lcrV196 expression was required for protection against challenge with Y. enterocolitica strain 8081, but was not sufficient to achieve significant protection against challenge with Y. enterocolitica strain WA, which expressed a divergent form of lcrV. Nevertheless, we are encouraged by these findings to continue pursuing our long-term goal of developing a single vaccine to protect against all three human pathogenic species of Yersinia.

Rapid Assembly of DNA via Ligase Cycling Reaction (LCR).

PubMed

Chandran, Sunil

2017-01-01

The assembly of multiple DNA parts into a larger DNA construct is a requirement in most synthetic biology laboratories. Here we describe a method for the efficient, high-throughput, assembly of DNA utilizing the ligase chain reaction (LCR). The LCR method utilizes non-overlapping DNA parts that are ligated together with the guidance of bridging oligos. Using this method, we have successfully assembled up to 20 DNA parts in a single reaction or DNA constructs up to 26 kb in size.

[Retrospective analysis of the value of arterial blood lactate level and its clearance rate on the prognosis of septic shock patients].

PubMed

Bao, Lei; Zhang, Min; Yan, Peixia; Wu, Xiaoyan; Shao, Jun; Zheng, Ruiqiang

2015-01-01

To explore the prognostic value of arterial blood lactate ( Lac ) levels and lactate clearance rate ( LCR ) in the patients with septic shock. A retrospective study was conducted. Clinical data of 94 septic patients admitted in the Department of Critical Care Medicine in Subei People's Hospital from January 2011 to June 2014 were analyzed. The arterial blood Lac levels at the moment of diagnosis of septic shock ( incipient value, 0 hour ) and early-stage after treatment ( 3, 6 and 24 hours ) were reviewed, and individual LCR was calculated at 3, 6, 24 hours for each patient. According to the outcome in intensive care unit ( ICU ), patients were divided into survival group ( n = 48 ) and death group ( n = 46 ). The Lac and LCR at different time points in two groups were analyzed, and the relationships between them and outcome were analyzed. The receiver-operating characteristic ( ROC ) curve was plotted to assess the value of Lac and LCR at different time points for predicting the outcome. Lac level after treatment in survival group was significantly lower than incipient value, but there was no obvious change in death group. Compared with death group, early Lac levels ( mmol/L ) in survival group were significantly reduced ( 0 hour: 3.80±2.14 vs. 5.75±3.21, 3 hours: 2.05±1.04 vs. 5.03±2.53, 6 hours: 1.80±0.77 vs. 4.40±2.02, 24 hours: 1.35±0.43 vs. 4.90±2.72, P<0.05 or P<0.01 ), the LCR was significantly increased [ 3 hours: 50.00 ( 72.35 )% vs. 13.51 ( 20.67 )%, 6 hours: 41.43 ( 58.42 )% vs. 22.00 ( 22.31 )%, 24 hours: 58.73 ( 29.94 )% vs. 18.92 ( 47.28 )%, P<0.05 or P<0.01 ]. The Lac levels at all time points were positively correlated with the outcome, and 6-hour and 24-hour LCR were negatively correlated with the outcome. According to the incipient Lac level, patients were divided into low Lac group ( Lac<2 mmol/L ), mild Lac group ( Lac 2-3 mmol/L ) and high Lac group ( Lac ≥ 4 mmol/L ). The mortality in low Lac group, mild Lac group, high Lac group was gradually increased [ 23.07% ( 6/26 ), 50.00% ( 8/16 ), 61.54% ( 32/52 ), χ(2) = 10.270, P = 0.006 ]. ROC curves demonstrated that the area under ROC curve ( AUC ) of 24-hour Lac was the largest, 0.944, and it was more sensitive and specific in the prognosis evaluation ( 100% and 78.3%, respectively ). According to the cut-off value of 24-hour Lac as 2.35 mmol/L, patients were divided into high Lac and low Lac groups, and mortality rate in high Lac group was significantly higher than that in low Lac group [ 100.0% ( 36/36 ) vs. 17.24% ( 10/58 ), χ(2) = 30.441,P = 0.000 ]. The AUC of 24-hour LCR was the largest, 0.865, and it was more sensitive and specific for the prognosis evaluation ( 83.3% and 91.3%, respectively ). According to the cut-off value of 24-hour LCR as 36.8%, patients were divided into high LCR group and low LCR group, and mortality rate in low LCR group was significantly higher than that in high LCR group [ 84.00% ( 42/50 ) vs. 9.09% ( 4/44 ), χ(2) = 26.278, P = 0.000 ]. Early high Lac in patients with septic shock prompts a poor prognosis, and 24-hour Lac levels and LCR are indicators of assessment of clinical therapeutic effect and prognosis of patients with septic shock.

Lunar Capabilities Roadmap

NASA Astrophysics Data System (ADS)

Kramer, G. Y.; Lawrence, D. J.; Neal, C. R.; Clark, P. E.; Green, R. O.; Horanyi, M.; Johnson, M. D.; Kelso, R. M.; Sultana, M.; Thompson, D. R.

2016-11-01

A Lunar Capabilities Roadmap (LCR) is required to highlight capabilities critical for science and exploration of the Moon as well as beyond. The LCR will focus mainly on capabilities with examples of specific technologies to satisfy those needs.

Systems-level computational modeling demonstrates fuel selection switching in high capacity running and low capacity running rats

PubMed Central

Qi, Nathan R.

2018-01-01

High capacity and low capacity running rats, HCR and LCR respectively, have been bred to represent two extremes of running endurance and have recently demonstrated disparities in fuel usage during transient aerobic exercise. HCR rats can maintain fatty acid (FA) utilization throughout the course of transient aerobic exercise whereas LCR rats rely predominantly on glucose utilization. We hypothesized that the difference between HCR and LCR fuel utilization could be explained by a difference in mitochondrial density. To test this hypothesis and to investigate mechanisms of fuel selection, we used a constraint-based kinetic analysis of whole-body metabolism to analyze transient exercise data from these rats. Our model analysis used a thermodynamically constrained kinetic framework that accounts for glycolysis, the TCA cycle, and mitochondrial FA transport and oxidation. The model can effectively match the observed relative rates of oxidation of glucose versus FA, as a function of ATP demand. In searching for the minimal differences required to explain metabolic function in HCR versus LCR rats, it was determined that the whole-body metabolic phenotype of LCR, compared to the HCR, could be explained by a ~50% reduction in total mitochondrial activity with an additional 5-fold reduction in mitochondrial FA transport activity. Finally, we postulate that over sustained periods of exercise that LCR can partly overcome the initial deficit in FA catabolic activity by upregulating FA transport and/or oxidation processes. PMID:29474500

LCR 5' hypersensitive site specificity for globin gene activation within the active chromatin hub.

PubMed

Peterson, Kenneth R; Fedosyuk, Halyna; Harju-Baker, Susanna

2012-12-01

The DNaseI hypersensitive sites (HSs) of the human β-globin locus control region (LCR) may function as part of an LCR holocomplex within a larger active chromatin hub (ACH). Differential activation of the globin genes during development may be controlled in part by preferential interaction of each gene with specific individual HSs during globin gene switching, a change in conformation of the LCR holocomplex, or both. To distinguish between these possibilities, human β-globin locus yeast artificial chromosome (β-YAC) lines were produced in which the ε-globin gene was replaced with a second marked β-globin gene (β(m)), coupled to an intact LCR, a 5'HS3 complete deletion (5'ΔHS3) or a 5'HS3 core deletion (5'ΔHS3c). The 5'ΔHS3c mice expressed β(m)-globin throughout development; γ-globin was co-expressed in the embryonic yolk sac, but not in the fetal liver; and wild-type β-globin was co-expressed in adult mice. Although the 5'HS3 core was not required for β(m)-globin expression, previous work showed that the 5'HS3 core is necessary for ε-globin expression during embryonic erythropoiesis. A similar phenotype was observed in 5'HS complete deletion mice, except β(m)-globin expression was higher during primitive erythropoiesis and γ-globin expression continued into fetal definitive erythropoiesis. These data support a site specificity model of LCR HS-globin gene interaction.

Genetic variations of E6 and long control region of human papillomavirus type 16 from patients with cervical lesion in Liaoning, China.

PubMed

Sun, Zhengrong; Lu, Zhitao; Liu, Jianhua; Wang, Guili; Zhou, Weiqiang; Yang, Lianxia; Liu, Chao; Wang, Bo; Ruan, Qiang

2013-10-07

High-risk human papillomavirus type 16 (HPV16) is a risk factor for cervical cancer. Previous studies suggest that polymorphisms in the E6 gene or the long control region(LCR)of HPV16 may alter the oncogenic potential of the virus. The aims of this study were to investigate the genetic variations of HPV16 E6 gene and LCR in isolates from Chinese population and correlation of the E6 and LCR polymorphisms with disease status of infected patients. HPV16 positive endocervical specimens were collected from 304 women living in Northeast of China. Sequences of E6 gene and LCR were analyzed by PCR-sequencing. Two lineages were found in the populations, including EUR lineage and As lineage. Based on the HPV16 prototype, the most frequent variation in the E6 gene was T178A/G (48.7%), followed by mutations of G94A (12.2%) and T350G (9.9%). The rank orders of incidence of E6 variations in amino acid were as follows: D25E (46.3%), L83V (9.9%) and H78Y (4.3%). Nucleotide variations in LCR were found in all the 304 isolates from HPV16 positive cervical samples. The most commonly observed LCR variations were the transition replacement G7193T, 7434CIns, G7521A and 7863ADel (100%). The As lineage was associated with HPV persistent infections and with disease status of ≥CIN2,3. The EUR lineage variants showed a negative trend of association with the severity of ≥CIN2,3. Among 41 variations found in LCR, 25 (61.0%) were located at the binding sites for transcription factors. Occurrence of ≥CIN2,3 was significantly associated with the mutations of R10G/L83V in E6 and the C7294T co-variation in LCR, after adjusting for ages of infected patients. Associations between As lineage and HPV persistent infections, and with disease status of ≥CIN2,3, and an association between the EUR lineage and negative trend of association with the severity of ≥CIN2,3 were found in this study. An association between a co-variation of R10G/L83V in E6 and C7294T in LCR and an increased risk for developing CIN-2,3 was found in a HPV16 infected population of Chinese women. These findings indicate that HPV16 polymorphism influences development of CIN-2,3.

Aerobic capacity mediates susceptibility for the transition from steatosis to steatohepatitis.

PubMed

Morris, E Matthew; McCoin, Colin S; Allen, Julie A; Gastecki, Michelle L; Koch, Lauren G; Britton, Steven L; Fletcher, Justin A; Fu, Xiarong; Ding, Wen-Xing; Burgess, Shawn C; Rector, R Scott; Thyfault, John P

2017-07-15

Low intrinsic aerobic capacity is associated with increased all-cause and liver-related mortality in humans. Low intrinsic aerobic capacity in the low capacity runner (LCR) rat increases susceptibility to acute and chronic high-fat/high-sucrose diet-induced steatosis, without observed increases in liver inflammation. Addition of excess cholesterol to a high-fat/high-sucrose diet produced greater steatosis in LCR and high capacity runner (HCR) rats. However, the LCR rat demonstrated greater susceptibility to increased liver inflammatory and apoptotic markers compared to the HCR rat. The progressive non-alcoholic fatty liver disease observed in the LCR rats following western diet feeding was associated with further declines in liver fatty acid oxidation and mitochondrial respiratory capacity compared to HCR rats. Low aerobic capacity increases risk for non-alcoholic fatty liver disease and liver-related disease mortality, but mechanisms mediating these effects remain unknown. We recently reported that rats bred for low aerobic capacity (low capacity runner; LCR) displayed susceptibility to high fat diet-induced steatosis in association with reduced hepatic mitochondrial fatty acid oxidation (FAO) and respiratory capacity compared to high aerobic capacity (high capacity runner; HCR) rats. Here we tested the impact of aerobic capacity on susceptibility for progressive liver disease following a 16-week 'western diet' (WD) high in fat (45% kcal), cholesterol (1% w/w) and sucrose (15% kcal). Unlike previously with a diet high in fat and sucrose alone, the inclusion of cholesterol in the WD induced hepatomegaly and steatosis in both HCR and LCR rats, while producing greater cholesterol ester accumulation in LCR compared to HCR rats. Importantly, WD-fed low-fitness LCR rats displayed greater inflammatory cell infiltration, serum alanine transaminase, expression of hepatic inflammatory markers (F4/80, MCP-1, TLR4, TLR2 and IL-1β) and effector caspase (caspase 3 and 7) activation compared to HCR rats. Further, LCR rats had greater WD-induced decreases in complete FAO and mitochondrial respiratory capacity. Intrinsic aerobic capacity had no impact on WD-induced hepatic steatosis; however, rats bred for low aerobic capacity developed greater hepatic inflammation, which was associated with reduced hepatic mitochondrial FAO and respiratory capacity and increased accumulation of cholesterol esters. These results confirm epidemiological reports that aerobic capacity impacts progression of liver disease and suggest that these effects are mediated through alterations in hepatic mitochondrial function. © 2017 The Authors. The Journal of Physiology © 2017 The Physiological Society.

Extrachromosomal HPV-16 LCR transcriptional activation by HDACi opposed by cellular differentiation and DNA integration.

PubMed

Bojilova, Ekaterina Dimitrova; Weyn, Christine; Antoine, Marie-Hélène; Fontaine, Véronique

2016-11-15

Histone deacetylase inhibitors (HDACi) have been shown to render HPV-carrying cells susceptible to intrinsic and extrinsic apoptotic signals. As such, these epigenetic drugs have entered clinical trials in the effort to treat cervical cancer. Here, we studied the effect of common HDACi, with an emphasis on Trichostatin A (TSA), on the transcriptional activity of the HPV-16 Long Control Region (LCR) in order to better understand the impact of these agents in the context of the HPV life cycle and infection. HDACi strongly induced transcription of the firefly luciferase reporter gene under the control of the HPV-16 LCR in a variety of cell lines. In the HaCaT keratinocyte cell line undergoing differentiation induced by TSA, we observed a reduction in LCR-controlled transcription. Three major AP-1 binding sites in the HPV-16 LCR are involved in the regulation by TSA. However, whatever the status of differentiation of the HaCaT cells, TSA induced integration of extra-chromosomal transfected DNA into the cellular genome. Although these data suggest caution using HDACi in the treatment of HR HPV infection, further in vivo studies are necessary to better assess the risk.

Extrachromosomal HPV-16 LCR transcriptional activation by HDACi opposed by cellular differentiation and DNA integration

PubMed Central

Bojilova, Ekaterina Dimitrova; Weyn, Christine; Antoine, Marie-Hélène; Fontaine, Véronique

2016-01-01

Histone deacetylase inhibitors (HDACi) have been shown to render HPV-carrying cells susceptible to intrinsic and extrinsic apoptotic signals. As such, these epigenetic drugs have entered clinical trials in the effort to treat cervical cancer. Here, we studied the effect of common HDACi, with an emphasis on Trichostatin A (TSA), on the transcriptional activity of the HPV-16 Long Control Region (LCR) in order to better understand the impact of these agents in the context of the HPV life cycle and infection. HDACi strongly induced transcription of the firefly luciferase reporter gene under the control of the HPV-16 LCR in a variety of cell lines. In the HaCaT keratinocyte cell line undergoing differentiation induced by TSA, we observed a reduction in LCR-controlled transcription. Three major AP-1 binding sites in the HPV-16 LCR are involved in the regulation by TSA. However, whatever the status of differentiation of the HaCaT cells, TSA induced integration of extra-chromosomal transfected DNA into the cellular genome. Although these data suggest caution using HDACi in the treatment of HR HPV infection, further in vivo studies are necessary to better assess the risk. PMID:27705914

Physical Activity Differentially Affects the Cecal Microbiota of Ovariectomized Female Rats Selectively Bred for High and Low Aerobic Capacity

PubMed Central

Liu, Tzu-Wen; Park, Young-Min; Holscher, Hannah D.; Padilla, Jaume; Scroggins, Rebecca J.; Welly, Rebecca; Britton, Steven L.; Koch, Lauren G.; Vieira-Potter, Victoria J.; Swanson, Kelly S.

2015-01-01

The gut microbiota is considered a relevant factor in obesity and associated metabolic diseases, for which postmenopausal women are particularly at risk. Increasing physical activity has been recognized as an efficacious approach to prevent or treat obesity, yet the impact of physical activity on the microbiota remains under-investigated. We examined the impacts of voluntary exercise on host metabolism and gut microbiota in ovariectomized (OVX) high capacity (HCR) and low capacity running (LCR) rats. HCR and LCR rats (age = 27wk) were OVX and fed a high-fat diet (45% kcal fat) ad libitum and housed in cages equipped with (exercise, EX) or without (sedentary, SED) running wheels for 11wk (n = 7-8/group). We hypothesized that increased physical activity would hinder weight gain, increase metabolic health and shift the microbiota of LCR rats, resulting in populations more similar to that of HCR rats. Animals were compared for characteristic metabolic parameters including body composition, lipid profile and energy expenditure; whereas cecal digesta were collected for DNA extraction. 16S rRNA gene-based amplicon Illumina MiSeq sequencing was performed, followed by analysis using QIIME 1.8.0 to assess cecal microbiota. Voluntary exercise decreased body and fat mass, and normalized fasting NEFA concentrations of LCR rats, despite only running one-third the distance of HCR rats. Exercise, however, increased food intake, weight gain and fat mass of HCR rats. Exercise clustered the gut microbial community of LCR rats, which separated them from the other groups. Assessments of specific taxa revealed significant (p<0.05) line by exercise interactions including shifts in the abundances of Firmicutes, Proteobacteria, and Cyanobacteria. Relative abundance of Christensenellaceae family was higher (p = 0.026) in HCR than LCR rats, and positively correlated (p<0.05) with food intake, body weight and running distance. These findings demonstrate that exercise differentially impacts host metabolism and gut microbial communities of female HCR and LCR rats without ovarian function. PMID:26301712

DNAzyme based gap-LCR detection of single-nucleotide polymorphism.

PubMed

Zhou, Li; Du, Feng; Zhao, Yongyun; Yameen, Afshan; Chen, Haodong; Tang, Zhuo

2013-07-15

Fast and accurate detection of single-nucleotide polymorphism (SNP) is thought more and more important for understanding of human physiology and elucidating the molecular based diseases. A great deal of effort has been devoted to developing accurate, rapid, and cost-effective technologies for SNP analysis. However most of those methods developed to date incorporate complicated probe labeling and depend on advanced equipment. The DNAzyme based Gap-LCR detection method averts any chemical modification on probes and circumvents those problems by incorporating a short functional DNA sequence into one of LCR primers. Two kinds of exonuclease are utilized in our strategy to digest all the unreacted probes and release the DNAzymes embedded in the LCR product. The DNAzyme applied in our method is a versatile tool to report the result of SNP detection in colorimetric or fluorometric ways for different detection purposes. Copyright © 2013 Elsevier B.V. All rights reserved.

LCR 5′ hypersensitive site specificity for globin gene activation within the active chromatin hub

PubMed Central

Peterson, Kenneth R.; Fedosyuk, Halyna; Harju-Baker, Susanna

2012-01-01

The DNaseI hypersensitive sites (HSs) of the human β-globin locus control region (LCR) may function as part of an LCR holocomplex within a larger active chromatin hub (ACH). Differential activation of the globin genes during development may be controlled in part by preferential interaction of each gene with specific individual HSs during globin gene switching, a change in conformation of the LCR holocomplex, or both. To distinguish between these possibilities, human β-globin locus yeast artificial chromosome (β-YAC) lines were produced in which the ε-globin gene was replaced with a second marked β-globin gene (βm), coupled to an intact LCR, a 5′HS3 complete deletion (5′ΔHS3) or a 5′HS3 core deletion (5′ΔHS3c). The 5′ΔHS3c mice expressed βm-globin throughout development; γ-globin was co-expressed in the embryonic yolk sac, but not in the fetal liver; and wild-type β-globin was co-expressed in adult mice. Although the 5′HS3 core was not required for βm-globin expression, previous work showed that the 5′HS3 core is necessary for ε-globin expression during embryonic erythropoiesis. A similar phenotype was observed in 5′HS complete deletion mice, except βm-globin expression was higher during primitive erythropoiesis and γ-globin expression continued into fetal definitive erythropoiesis. These data support a site specificity model of LCR HS-globin gene interaction. PMID:23042246

Activation of beta-major globin gene transcription is associated with recruitment of NF-E2 to the beta-globin LCR and gene promoter.

PubMed

Sawado, T; Igarashi, K; Groudine, M

2001-08-28

The mouse beta-globin gene locus control region (LCR), located upstream of the beta-globin gene cluster, is essential for the activated transcription of genes in the cluster. The LCR contains multiple binding sites for transactivators, including Maf-recognition elements (MAREs). However, little is known about the specific proteins that bind to these sites or the time at which they bind during erythroid differentiation. We have performed chromatin immunoprecipitation experiments to determine the recruitment of the erythroid-specific transactivator p45 NF-E2/MafK (p18 NF-E2) heterodimer and small Maf proteins to various regions in the globin gene locus before and after the induction of murine erythroleukemia (MEL) cell differentiation. We report that, before induction, the LCR is occupied by small Maf proteins, and, on erythroid maturation, the NF-E2 complex is recruited to the LCR and the active globin promoters, even though the promoters do not contain MAREs. This differentiation-coupled recruitment of NF-E2 complex correlates with a greater than 100-fold increase in beta-major globin transcription, but is not associated with a significant change in locus-wide histone H3 acetylation. These findings suggest that the beta-globin gene locus exists in a constitutively open chromatin conformation before terminal differentiation, and we speculate that recruitment of NF-E2 complex to the LCR and active promoters may be a rate-limiting step in the activation of beta-globin gene expression.

Distribution and movement of humpback chub in the Colorado River, Grand Canyon, based on recaptures

USGS Publications Warehouse

Paukert, C.P.; Coggins, L.G.; Flaccus, C.E.

2006-01-01

Mark-recapture data from the federally endangered humpback chub Gila cypha in the Colorado River, Grand Canyon, were analyzed from 1989 to 2002 to determine large-scale movement patterns and distribution. A total of 14,674 recaptures from 7,127 unique fish were documented; 87% of the recaptures occurred in the same main-stem river reach or tributary as the original captures, suggesting restricted distribution by most fish. A total of 99% of all recaptures were from in and around the Little Colorado River (LCR), a tributary of the Colorado River and primary aggregation and spawning location of humpback chub in Grand Canyon. Time at liberty averaged 394 d, but some fish were recaptured near their main-stem capture location over 10 years later. Proportionally fewer large (>300-mm) humpback chub exhibited restricted distribution than small (<200-mm) fish. However, several fish did move more than 154 km throughout Grand Canyon between capture and recapture, suggesting that limited movement occurs throughout Grand Canyon. The majority of the recaptured fish remained in or returned to the LCR or the Colorado River near the LCR. Although many large-river fishes exhibit extensive migrations to fulfill their life history requirements, most of the humpback chub in Grand Canyon appear to remain in or come back to the LCR and LCR confluence across multiple sizes and time scales. Detecting trends in the overall abundance of this endangered fish in Grand Canyon can probably be accomplished by monitoring the area in and around the LCR.

Heparin (GAG-hed) inhibits LCR activity of human papillomavirus type 18 by decreasing AP1 binding.

PubMed

Villanueva, Rita; Morales-Peza, Néstor; Castelán-Sánchez, Irma; García-Villa, Enrique; Tapia, Rocio; Cid-Arregui, Angel; García-Carrancá, Alejandro; López-Bayghen, Esther; Gariglio, Patricio

2006-08-31

High risk HPVs are causative agents of anogenital cancers. Viral E6 and E7 genes are continuously expressed and are largely responsible for the oncogenic activity of these viruses. Transcription of the E6 and E7 genes is controlled by the viral Long Control Region (LCR), plus several cellular transcription factors including AP1 and the viral protein E2. Within the LCR, the binding and activity of the transcription factor AP1 represents a key regulatory event in maintaining E6/E7 gene expression and uncontrolled cell proliferation. Glycosaminoglycans (GAGs), such as heparin, can inhibit tumour growth; they have also shown antiviral effects and inhibition of AP1 transcriptional activity. The purpose of this study was to test the heparinoid GAG-hed, as a possible antiviral and antitumoral agent in an HPV18 positive HeLa cell line. Using in vivo and in vitro approaches we tested GAG-hed effects on HeLa tumour cell growth, cell proliferation and on the expression of HPV18 E6/E7 oncogenes. GAG-hed effects on AP1 binding to HPV18-LCR-DNA were tested by EMSA. We were able to record the antitumoral effect of GAG-hed in vivo by using as a model tumours induced by injection of HeLa cells into athymic female mice. The antiviral effect of GAG-hed resulted in the inhibition of LCR activity and, consequently, the inhibition of E6 and E7 transcription. A specific diminishing of cell proliferation rates was observed in HeLa but not in HPV-free colorectal adenocarcinoma cells. Treated HeLa cells did not undergo apoptosis but the percentage of cells in G2/M phase of the cell cycle was increased. We also detected that GAG-hed prevents the binding of the transcription factor AP1 to the LCR. Direct interaction of GAG-hed with the components of the AP1 complex and subsequent interference with its ability to correctly bind specific sites within the viral LCR may contribute to the inhibition of E6/E7 transcription and cell proliferation. Our data suggest that GAG-hed could have antitumoral and antiviral activity mainly by inhibiting AP1 binding to the HPV18-LCR.

Life/Career Renewal: An Intervention for Vocational and Other Life Transitions.

ERIC Educational Resources Information Center

Robitschek, Christine

1997-01-01

A study examined the effectiveness of the Life/Career Renewal Program (LCR), a wilderness program for adults, in enhancing personal growth initiative (PGI) of 68 participants. Results support the hypothesis that the LCR program enhances PGI. (Author/JOW)

Implementation of the qualities of radiodiagnostic: mammography

NASA Astrophysics Data System (ADS)

Pacífico, L. C.; Magalhães, L. A. G.; Peixoto, J. G. P.; Fernandes, E.

2018-03-01

The objective of the present study was to evaluate the expanded uncertainty of the mammographic calibration process and present the result of the internal audit performed at the Laboratory of Radiological Sciences (LCR). The qualities of the mammographic beans that are references in the LCR, comprises two irradiation conditions: no-attenuated beam and attenuated beam. Both had satisfactory results, with an expanded uncertainty equals 2,1%. The internal audit was performed, and the degree of accordance with the ISO/IEC 17025 was evaluated. The result of the internal audit was satisfactory. We conclude that LCR can perform calibrations on mammography qualities for end users.

Long-range looping of a locus control region drives tissue-specific chromatin packing within a multigene cluster

PubMed Central

Tsai, Yu-Cheng; Cooke, Nancy E.; Liebhaber, Stephen A.

2016-01-01

Abstract The relationships of higher order chromatin organization to mammalian gene expression remain incompletely defined. The human Growth Hormone (hGH) multigene cluster contains five gene paralogs. These genes are selectively activated in either the pituitary or the placenta by distinct components of a remote locus control region (LCR). Prior studies have revealed that appropriate activation of the placental genes is dependent not only on the actions of the LCR, but also on the multigene composition of the cluster itself. Here, we demonstrate that the hGH LCR ‘loops’ over a distance of 28 kb in primary placental nuclei to make specific contacts with the promoters of the two GH genes in the cluster. This long-range interaction sequesters the GH genes from the three hCS genes which co-assemble into a tightly packed ‘hCS chromatin hub’. Elimination of the long-range looping, via specific deletion of the placental LCR components, triggers a dramatic disruption of the hCS chromatin hub. These data reveal a higher-order structural pathway by which long-range looping from an LCR impacts on local chromatin architecture that is linked to tissue-specific gene regulation within a multigene cluster. PMID:26893355

Region-specific differences in bioenergetic proteins and protein response to acute high fat diet in brains of low and high capacity runner rats.

PubMed

Gan, Li; Ma, Delin; Li, Min; Yang, Fu-Chen; Rogers, Robert S; Wheatley, Joshua L; Koch, Lauren G; Britton, Steven L; Thyfault, John P; Geiger, Paige C; Stanford, John A

2018-05-01

Aerobic capacity is a strong predictor of mortality. Low capacity runner (LCR) rats exhibit reduced mitochondrial function in peripheral organs. A high fat diet (HFD) can worsen metabolic phenotype in LCR rats. Little is known about metabolic changes in the brains of these rats, however. This study examined protein markers of mitochondrial function and metabolism as a function of aerobic running capacity and an acute HFD in four brain regions: the striatum, hippocampus, hypothalamus, and substantia nigra. After 3 days HFD or chow diets, we measured peroxisome proliferator-activated receptor-γ coactivator 1α (PGC1-α), nuclear respiratory factors 1 (Nrf-1), mitochondrial transcription factor A (TFAM), and phosphorylated (activated) AMP-activated protein kinase (p-AMPK) protein levels in the four brain regions. LCR rats exhibited lower levels of mitochondrial proteins (PGC1-α, Nrf-1, TFAM), and greater p-AMPK, in striatum, but not in the other brain regions. Mitochondrial protein levels were greater in HFD LCR striatum, while p-AMPK was lower in this group. Markers of lower mitochondrial biogenesis and increased metabolic demand were limited to the LCR striatum, which nevertheless maintained the capacity to respond to an acute HFD challenge. Copyright © 2018 Elsevier B.V. All rights reserved.

Efficacy and safety profile of LCR35 complete freeze-dried culture in irritable bowel syndrome: a randomized, double-blind study.

PubMed

Dapoigny, Michel; Piche, Thierry; Ducrotte, Philippe; Lunaud, Bernard; Cardot, Jean-Michel; Bernalier-Donadille, Annick

2012-05-07

To assess the effects and safety of Lactobacillus casei rhamnosus LCR35 complete freeze-dried culture (LCR35) in patients suffering from irritable bowel syndrome (IBS). A randomized, double-blind pilot study was performed in 50 patients complaining of IBS symptoms complying with Rome III criteria. Patients were allocated to receive either LCR35 (n = 25) at a minimum daily dose of 6 × 10(8) colony forming units or placebo (n = 25) for 4 wk. At inclusion, after treatment and 2 wk later, patients completed the IBS severity scale. Change from baseline in the IBS severity score at the end of treatment was the primary efficacy criterion. Changes were compared between groups in the whole population and in IBS subtypes (IBS with predominance of constipation, IBS with predominance of diarrhoea, mixed IBS, unsubtyped IBS). The presence of lactobacillus casei rhamnosus in stools was investigated at inclusion and at the end of treatment. The gastrointestinal quality of life questionnaire and the hospital anxiety and depression (HAD) scale were also completed. Both groups were balanced for baseline characteristics. In 85% of patients, stool analyses showed that lactobacillus casei rhamnosus able to survive in the digestive tract. In the whole population, improvements in the IBS severity score did not differ significantly between treatments with a 25% decrease after 4-wk treatment, and a 15% decrease from baseline 2 wk later in both groups. In IBS subgroups, statistical analysis could not be performed due to small sample size, but a clinical response in favour of LCR35 was observed in IBS patients with predominance of diarrhoea: no change in the symptom severity score was seen with the placebo after 4 wk treatment, whereas a clinically relevant decrease occurred with LCR35 (-37% vs -3%). Furthermore, in spite of an increase in symptom intensity, the IBS severity score was maintained below the baseline value 2 wk later with LCR35 (-19% from baseline), whilst a slight 5% increase from baseline was observed with placebo. In the IBS subgroup with predominance of diarrhoea only, a clinically relevant decrease in abdominal pain severity score (-36%) was observed with LCR35, whereas no change occurred with placebo. In mixed IBS patients, the 20% and 30% decreases in the IBS severity score observed after treatment with LCR35 and placebo, respectively, were maintained 2 wk later in both groups. A clinical response slightly in favour of placebo was observed at the end of the treatment period in IBS patients with predominance of constipation (-41% vs -20%) and unsubtyped IBS patients (-47% vs -17%), with the same value maintained 2 wk later. In both groups, no clinically relevant changes were observed either for the gastrointestinal quality of life index or HAD score. Thus, these results suggest that sub-grouping of IBS patients may be important for optimizing treatment responses by the physician. This pilot study suggests that LCR35 could have some efficacy in IBS patients complaining of diarrhoea. These preliminary results need to be confirmed in larger studies.

Two Decades into the LCR: What We Do and Still Don’t Know to Solve Lead Problems

EPA Science Inventory

Site selection and sampling protocol biases in LCR samplingunderestimate peak lead and copper concentrations whilemissing erratic lead release episodes resulting from distributionsystem chemical and physical disturbances. Possible sitetargeting and sampling protocol changes could...

Two Decades into the LCR: What We Do and Still Don’t Know to Solve Lead Problems - abstract

EPA Science Inventory

Site selection and sampling protocol biases in LCR samplingunderestimate peak lead and copper concentrations whilemissing erratic lead release episodes resulting from distributionsystem chemical and physical disturbances. Possible sitetargeting and sampling protocol changes could...

Application of LCR Waves to Inspect Aircraft Structures

DTIC Science & Technology

2013-01-01

Mechanical Engineering (COBEM 2011). Proceedings of COBEM, 2011. Natal, RN, Brasil Analysis of the behavior of Lcr Waves propagating in Steel bars using...Taguchi Method. 21 th International Congress of Mechanical Engineering (COBEM 2011). Proceedings of COBEM, 2011. Natal, RN, Brasil . Application

Eating habits modulate short term memory and epigenetical regulation of brain derived neurotrophic factor in hippocampus of low- and high running capacity rats.

PubMed

Torma, Ferenc; Bori, Zoltan; Koltai, Erika; Felszeghy, Klara; Vacz, Gabriella; Koch, Lauren; Britton, Steven; Boldogh, Istvan; Radak, Zsolt

2014-08-01

Exercise capacity and dietary restriction (DR) are linked to improved quality of life, including enhanced brain function and neuro-protection. Brain derived neurotrophic factor (BDNF) is one of the key proteins involved in the beneficial effects of exercise on brain. Low capacity runner (LCR) and high capacity runner (HCR) rats were subjected to DR in order to investigate the regulation of BDNF. HCR-DR rats out-performed other groups in a passive avoidance test. BDNF content increased significantly in the hippocampus of HCR-DR groups compared to control groups (p<0.05). The acetylation of H3 increased significantly only in the LCR-DR group. However, chip-assay revealed that the specific binding between acetylated histone H3 and BNDF promoter was increased in both LCR-DR and HCR-DR groups. In spite of these increases in binding, at the transcriptional level only, the LCR-DR group showed an increase in BDNF mRNA content. Additionally, DR also induced the activity of cAMP response element-binding protein (CREB), while the content of SIRT1 was not altered. Peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α) was elevated in HCR-DR groups. But, based on the levels of nuclear respiratory factor-1 and cytocrome c oxidase, it appears that DR did not cause mitochondrial biogenesis. The data suggest that DR-mediated induction of BDNF levels includes chromatin remodeling. Moreover, DR does not induce mitochondrial biogenesis in the hippocampus of LCR/HCR rats. DR results in different responses to a passive avoidance test, and BDNF regulation in LCR and HCR rats. Copyright © 2014 Elsevier Inc. All rights reserved.

Effects of ovariectomy and intrinsic aerobic capacity on tissue-specific insulin sensitivity

PubMed Central

Park, Young-Min; Rector, R. Scott; Thyfault, John P.; Zidon, Terese M.; Padilla, Jaume; Welly, Rebecca J.; Meers, Grace M.; Morris, Matthew E.; Britton, Steven L.; Koch, Lauren G.; Booth, Frank W.; Kanaley, Jill A.

2015-01-01

High-capacity running (HCR) rats are protected against the early (i.e., ∼11 wk postsurgery) development of ovariectomy (OVX)-induced insulin resistance (IR) compared with low-capacity running (LCR) rats. The purpose of this study was to utilize the hyperinsulinemic euglycemic clamp to determine whether 1) HCR rats remain protected from OVX-induced IR when the time following OVX is extended to 27 wk and 2) tissue-specific glucose uptake differences are responsible for the protection in HCR rats under sedentary conditions. Female HCR and LCR rats (n = 40; aged ∼22 wk) randomly received either OVX or sham (SHM) surgeries and then underwent the clamp 27 wk following surgeries. [3-3H]glucose was used to determine glucose clearance, whereas 2-[14C]deoxyglucose (2-DG) was used to assess glucose uptake in skeletal muscle, brown adipose tissue (BAT), subcutaneous white adipose tissue (WAT), and visceral WAT. OVX decreased the glucose infusion rate and glucose clearance in both lines, but HCR had better insulin sensitivity than LCR (P < 0.05). In both lines, OVX significantly reduced glucose uptake in soleus and gastrocnemius muscles; however, HCR showed ∼40% greater gastrocnemius glucose uptake compared with LCR (P < 0.05). HCR also exhibited greater glucose uptake in BAT and visceral WAT compared with LCR (P < 0.05), yet these tissues were not affected by OVX in either line. In conclusion, OVX impairs insulin sensitivity in both HCR and LCR rats, likely driven by impairments in insulin-mediated skeletal muscle glucose uptake. HCR rats have greater skeletal muscle, BAT, and WAT insulin-mediated glucose uptake, which may aid in protection against OVX-associated insulin resistance. PMID:26646101

Female rats selectively bred for high intrinsic aerobic fitness are protected from ovariectomy-associated metabolic dysfunction

PubMed Central

Padilla, Jaume; Park, Young-Min; Welly, Rebecca J.; Scroggins, Rebecca J.; Britton, Steven L.; Koch, Lauren G.; Jenkins, Nathan T.; Crissey, Jacqueline M.; Zidon, Terese; Morris, E. Matthew; Meers, Grace M. E.; Thyfault, John P.

2015-01-01

Ovariectomized rodents model human menopause in that they rapidly gain weight, reduce spontaneous physical activity (SPA), and develop metabolic dysfunction, including insulin resistance. How contrasting aerobic fitness levels impacts ovariectomy (OVX)-associated metabolic dysfunction is not known. Female rats selectively bred for high and low intrinsic aerobic fitness [high-capacity runners (HCR) and low-capacity runners (LCR), respectively] were maintained under sedentary conditions for 39 wk. Midway through the observation period, OVX or sham (SHM) operations were performed providing HCR-SHM, HCR-OVX, LCR-SHM, and LCR-OVX groups. Glucose tolerance, energy expenditure, and SPA were measured before and 4 wk after surgery, while body composition via dual-energy X-ray absorptiometry and adipose tissue distribution, brown adipose tissue (BAT), and skeletal muscle phenotype, hepatic lipid content, insulin resistance via homeostatic assessment model of insulin resistance and AdipoIR, and blood lipids were assessed at death. Remarkably, HCR were protected from OVX-associated increases in adiposity and insulin resistance, observed only in LCR. HCR rats were ∼30% smaller, had ∼70% greater spontaneous physical activity (SPA), consumed ∼10% more relative energy, had greater skeletal muscle proliferator-activated receptor coactivator 1-alpha, and ∼40% more BAT. OVX did not increase energy intake and reduced SPA to the same extent in both HCR and LCR. LCR were particularly affected by an OVX-associated reduction in resting energy expenditure and experienced a reduction in relative BAT; resting energy expenditure correlated positively with BAT across all animals (r = 0.6; P < 0.001). In conclusion, despite reduced SPA following OVX, high intrinsic aerobic fitness protects against OVX-associated increases in adiposity and insulin resistance. The mechanism may involve preservation of resting energy expenditure. PMID:25608751

Novel quantitative real-time LCR for the sensitive detection of SNP frequencies in pooled DNA: method development, evaluation and application.

PubMed

Psifidi, Androniki; Dovas, Chrysostomos; Banos, Georgios

2011-01-19

Single nucleotide polymorphisms (SNP) have proven to be powerful genetic markers for genetic applications in medicine, life science and agriculture. A variety of methods exist for SNP detection but few can quantify SNP frequencies when the mutated DNA molecules correspond to a small fraction of the wild-type DNA. Furthermore, there is no generally accepted gold standard for SNP quantification, and, in general, currently applied methods give inconsistent results in selected cohorts. In the present study we sought to develop a novel method for accurate detection and quantification of SNP in DNA pooled samples. The development and evaluation of a novel Ligase Chain Reaction (LCR) protocol that uses a DNA-specific fluorescent dye to allow quantitative real-time analysis is described. Different reaction components and thermocycling parameters affecting the efficiency and specificity of LCR were examined. Several protocols, including gap-LCR modifications, were evaluated using plasmid standard and genomic DNA pools. A protocol of choice was identified and applied for the quantification of a polymorphism at codon 136 of the ovine PRNP gene that is associated with susceptibility to a transmissible spongiform encephalopathy in sheep. The real-time LCR protocol developed in the present study showed high sensitivity, accuracy, reproducibility and a wide dynamic range of SNP quantification in different DNA pools. The limits of detection and quantification of SNP frequencies were 0.085% and 0.35%, respectively. The proposed real-time LCR protocol is applicable when sensitive detection and accurate quantification of low copy number mutations in DNA pools is needed. Examples include oncogenes and tumour suppressor genes, infectious diseases, pathogenic bacteria, fungal species, viral mutants, drug resistance resulting from point mutations, and genetically modified organisms in food.

Novel Quantitative Real-Time LCR for the Sensitive Detection of SNP Frequencies in Pooled DNA: Method Development, Evaluation and Application

PubMed Central

Psifidi, Androniki; Dovas, Chrysostomos; Banos, Georgios

2011-01-01

Background Single nucleotide polymorphisms (SNP) have proven to be powerful genetic markers for genetic applications in medicine, life science and agriculture. A variety of methods exist for SNP detection but few can quantify SNP frequencies when the mutated DNA molecules correspond to a small fraction of the wild-type DNA. Furthermore, there is no generally accepted gold standard for SNP quantification, and, in general, currently applied methods give inconsistent results in selected cohorts. In the present study we sought to develop a novel method for accurate detection and quantification of SNP in DNA pooled samples. Methods The development and evaluation of a novel Ligase Chain Reaction (LCR) protocol that uses a DNA-specific fluorescent dye to allow quantitative real-time analysis is described. Different reaction components and thermocycling parameters affecting the efficiency and specificity of LCR were examined. Several protocols, including gap-LCR modifications, were evaluated using plasmid standard and genomic DNA pools. A protocol of choice was identified and applied for the quantification of a polymorphism at codon 136 of the ovine PRNP gene that is associated with susceptibility to a transmissible spongiform encephalopathy in sheep. Conclusions The real-time LCR protocol developed in the present study showed high sensitivity, accuracy, reproducibility and a wide dynamic range of SNP quantification in different DNA pools. The limits of detection and quantification of SNP frequencies were 0.085% and 0.35%, respectively. Significance The proposed real-time LCR protocol is applicable when sensitive detection and accurate quantification of low copy number mutations in DNA pools is needed. Examples include oncogenes and tumour suppressor genes, infectious diseases, pathogenic bacteria, fungal species, viral mutants, drug resistance resulting from point mutations, and genetically modified organisms in food. PMID:21283808

Laparoscopic colon resection: To prep or not to prep? Analysis of 1535 patients.

PubMed

Allaix, Marco Ettore; Arolfo, Simone; Degiuli, Maurizio; Giraudo, Giuseppe; Volpatto, Silvio; Morino, Mario

2016-06-01

Mechanical bowel preparation (MBP) before elective open colon resection does not reduce the rate of postoperative anastomotic leakage. However, MBP is still routinely used in many countries, and there are very limited data regarding the utility of preoperative MBP in patients undergoing laparoscopic colon resection (LCR). The aim of this study was to challenge the use of MBP before elective LCR. It is a retrospective analysis of a prospectively collected database. All patients undergoing elective LCR with primary anastomosis and no stoma were included. Preoperative MBP with polyethylene glycol solution was used routinely between April 1992 and December 2004, and then it was abandoned. The early postoperative outcomes in patients who had preoperative MBP (MBP group) and in patients who underwent LCR without preoperative MBP (No-MBP group) were compared. From April 1992 to December 2014, 1535 patients underwent LCR: 706 MBP patients and 829 No-MBP patients. There were no differences in demographic data, indication for surgery and type of procedure performed between MBP and No-MBP group patients. The incidence of anastomotic leakage was similar between the two groups (3.4 vs. 3.6 %, p = 0.925). No differences were observed in intra-abdominal abscesses (0.6 vs. 0.8 %, p = 0.734), wound infections (0.6 vs. 1.4 %, p = 0.149), infectious extra-abdominal complications (1.8 vs. 3 %, p = 0.190), and non-infectious complications (6.1 vs. 6.8 %, p = 0.672). The overall reoperation rate was 4.6 % for MBP patients and 5 % for No-MBP patients (p = 0.813). The use of preoperative MBP does not seem to be associated with lower incidence of intra-abdominal septic complications after LCR.

Specification of unique Pit-1 activity in the hGH locus control region

PubMed Central

Shewchuk, Brian M.; Liebhaber, Stephen A.; Cooke, Nancy E.

2002-01-01

The human GH (hGH) gene cluster is regulated by a remote 5′ locus control region (LCR). HSI, an LCR component located 14.5 kb 5′ to the hGH-N promoter, constitutes the primary determinant of high-level hGH-N activation in pituitary somatotropes. HSI encompasses an array of three binding sites for the pituitary-specific POU homeodomain factor Pit-1. In the present report we demonstrate that all three Pit-1 sites in the HSI array contribute to LCR activity in vivo. Furthermore, these three sites as a unit are fully sufficient for position-independent and somatotrope-restricted hGH-N transgene activation. In contrast, the hGH-N transgene is not activated by Pit-1 sites native to either the hGH-N or rat (r)GH gene promoters. These findings suggest that the structures of the Pit-1 binding sites at HSI specify distinct chromatin-dependent activities essential for LCR-mediated activation of hGH in the developing pituitary. PMID:12189206

Friction and Wear Characteristics of a Modified Composite Solid Lubricant Plasma Spray Coating

NASA Technical Reports Server (NTRS)

Stanford, M. K.; DellaCorte, C.

2004-01-01

LCR304 is a solid lubricant coating composed of Ni-10Cr, Cr2O3, BaF2-CaF2 and Ag and developed for dimensional stability in high temperature air. This coating is a modification of PS304, which differs in that the Ni-Cr constituent contains 20wt% Cr. The tribological characteristics of LCR304 were evaluated by pin-on-disk and foil air bearing rig testing from 25 to 650 C and compared to previous test results with PS304. For both tests, the friction coefficient decreased as temperature increased from 25 to 650 C. Wear generally decreased with increasing temperature for all pin-on-disk tests. LCR304 coated components produced the least wear of Inconel X-750 counterface materials at 427 and 650 C. These results indicate that the LCR304 coating has potential as a replacement for PS304 in, for example, low cycle (minimum wear) applications where dimensional stability is imperative.

Cosmogenic 3He age estimates of Plio-Pleistocene alluvial-fan surfaces in the Lower Colorado River Corridor, Arizona, USA

NASA Astrophysics Data System (ADS)

Fenton, Cassandra R.; Pelletier, Jon D.

2013-01-01

Plio-Pleistocene deposits of the Lower Colorado River (LCR) and tributary alluvial fans emanating from the Black Mountains near Golden Shores, Arizona record six cycles of Late Cenozoic aggradation and incision of the LCR and its adjacent alluvial fans. Cosmogenic 3He (3Hec) ages of basalt boulders on fan terraces yield age ranges of: 3.3-2.2 Ma, 2.2-1.1 Ma, 1.1 Ma to 110 ka, < 350 ka, < 150 ka, and < 63 ka. T1 and Q1 fans are especially significant, because they overlie Bullhead Alluvium, i.e. the first alluvial deposit of the LCR since its inception ca. 4.2 Ma. 3Hec data suggest that the LCR began downcutting into the Bullhead Alluvium as early as 3.3 Ma and as late as 2.2 Ma. Younger Q2a to Q4 fans very broadly correlate in number and age with alluvial terraces elsewhere in the southwestern USA. Large uncertainties in 3Hec ages preclude a temporal link between the genesis of the Black Mountain fans and specific climate transitions. Fan-terrace morphology and the absence of significant Plio-Quaternary faulting in the area, however, indicate regional, episodic increases in sediment supply, and that climate change has possibly played a role in Late Cenozoic piedmont and valley-floor aggradation in the LCR valley.

H3 K79 dimethylation marks developmental activation of the beta-globin gene but is reduced upon LCR-mediated high-level transcription.

PubMed

Sawado, Tomoyuki; Halow, Jessica; Im, Hogune; Ragoczy, Tobias; Bresnick, Emery H; Bender, M A; Groudine, Mark

2008-07-15

Genome-wide analyses of the relationship between H3 K79 dimethylation and transcription have revealed contradictory results. To clarify this relationship at a single locus, we analyzed expression and H3 K79 modification levels of wild-type (WT) and transcriptionally impaired beta-globin mutant genes during erythroid differentiation. Analysis of fractionated erythroid cells derived from WT/Delta locus control region (LCR) heterozygous mice reveals no significant H3 K79 dimethylation of the beta-globin gene on either allele prior to activation of transcription. Upon transcriptional activation, H3 K79 di-methylation is observed along both WT and DeltaLCR alleles, and both alleles are located in proximity to H3 K79 dimethylation nuclear foci. However, H3 K79 di-methylation is significantly increased along the DeltaLCR allele compared with the WT allele. In addition, analysis of a partial LCR deletion mutant reveals that H3 K79 dimethylation is inversely correlated with beta-globin gene expression levels. Thus, while our results support a link between H3 K79 dimethylation and gene expression, high levels of this mark are not essential for high level beta-globin gene transcription. We propose that H3 K79 dimethylation is destabilized on a highly transcribed template.

Structural polymorphism at LCR and its role in beta-globin gene regulation.

PubMed

Kukreti, Shrikant; Kaur, Harpreet; Kaushik, Mahima; Bansal, Aparna; Saxena, Sarika; Kaushik, Shikha; Kukreti, Ritushree

2010-09-01

Information on the secondary structures and conformational manifestations of eukaryotic DNA and their biological significance with reference to gene regulation and expression is limited. The human beta-globin gene Locus Control Region (LCR), a dominant regulator of globin gene expression, is a contiguous piece of DNA with five tissue-specific DNase I-hypersensitive sites (HSs). Since these HSs have a high density of transcription factor binding sites, structural interdependencies between HSs and different promoters may directly or indirectly regulate LCR functions. Mutations and SNPs may stabilize or destabilize the local secondary structures, affecting the gene expression by changes in the protein-DNA recognition patterns. Various palindromic or quasi-palindromic segments within LCR, could cause structural polymorphism and geometrical switching of DNA. This emphasizes the importance of understanding of the sequence-dependent variations of the DNA structure. Such structural motifs might act as regulatory elements. The local conformational variability of a DNA segment or action of a DNA specific protein is key to create and maintain active chromatin domains and affect transcription of various tissue specific beta-globin genes. We, summarize here the current status of beta-globin LCR structure and function. Further structural studies at molecular level and functional genomics might solve the regulatory puzzles that control the beta-globin gene locus. Copyright (c) 2010 Elsevier Masson SAS. All rights reserved.

Chromatin structure of the LCR in the human β-globin locus transcribing the adult δ- and β-globin genes.

PubMed

Kim, Seoyeon; Kim, Yea Woon; Shim, Sung Han; Kim, Chul Geun; Kim, Aeri

2012-03-01

The β-like globin genes are transcribed in a developmental stage specific fashion in erythroid cells. The specific transcription of globin genes is conferred by the locus control region (LCR), but the chromatin structure of the LCR in the human adult β-globin locus transcribing the δ- and β-globin genes is not clear. Here, we employed hybrid MEL cells that contain a human chromosome 11. The δ- and β-globin genes were highly transcribed in hybrid MEL/ch11 cells after transcriptional induction. LCR HS3 and HS2 were strongly occupied by erythroid specific transcriptional activators and co-factors in the induced locus. These HSs, but not HS4 and HS1, were in close proximity with the active globin genes as revealed by high resolution 3C experiments. The active features at HS3 were markedly established after transcriptional induction, while HS2 was in a relatively active conformation before the induction. Unexpectedly, HS1 did not show notable active features except histone hyperacetylation. Taken together, the LCR of the human β-globin locus transcribing the adult δ- and β-globin genes has HS specific chromatin structure. The structure at each HS, which is different from the locus transcribing the fetal globin genes, might relate to its role in transcribing the adult genes. Copyright © 2011 Elsevier Ltd. All rights reserved.

Zebrafish globin switching occurs in two developmental stages and is controlled by the LCR.

PubMed

Ganis, Jared J; Hsia, Nelson; Trompouki, Eirini; de Jong, Jill L O; DiBiase, Anthony; Lambert, Janelle S; Jia, Zhiying; Sabo, Peter J; Weaver, Molly; Sandstrom, Richard; Stamatoyannopoulos, John A; Zhou, Yi; Zon, Leonard I

2012-06-15

Globin gene switching is a complex, highly regulated process allowing expression of distinct globin genes at specific developmental stages. Here, for the first time, we have characterized all of the zebrafish globins based on the completed genomic sequence. Two distinct chromosomal loci, termed major (chromosome 3) and minor (chromosome 12), harbor the globin genes containing α/β pairs in a 5'-3' to 3'-5' orientation. Both these loci share synteny with the mammalian α-globin locus. Zebrafish globin expression was assayed during development and demonstrated two globin switches, similar to human development. A conserved regulatory element, the locus control region (LCR), was revealed by analyzing DNase I hypersensitive sites, H3K4 trimethylation marks and GATA1 binding sites. Surprisingly, the position of these sites with relation to the globin genes is evolutionarily conserved, despite a lack of overall sequence conservation. Motifs within the zebrafish LCR include CACCC, GATA, and NFE2 sites, suggesting functional interactions with known transcription factors but not the same LCR architecture. Functional homology to the mammalian α-LCR MCS-R2 region was confirmed by robust and specific reporter expression in erythrocytes of transgenic zebrafish. Our studies provide a comprehensive characterization of the zebrafish globin loci and clarify the regulation of globin switching. Copyright © 2012 Elsevier Inc. All rights reserved.

The polymorphisms of LCR, E6, and E7 of HPV-58 isolates in Yunnan, Southwest China.

PubMed

Xi, Juemin; Chen, Junying; Xu, Miaoling; Yang, Hongying; Wen, Songjiao; Pan, Yue; Wang, Xiaodan; Ye, Chao; Qiu, Lijuan; Sun, Qiangming

2018-04-25

Variations in HPV LCR/E6/E7 have been shown to be associated with the viral persistence and cervical cancer development. So far, there are few reports about the polymorphisms of the HPV-58 LCR/E6/E7 sequences in Southwest China. This study aims to characterize the gene polymorphisms of the HPV-58 LCR/E6/E7 sequences in women of Southwest China, and assess the effects of variations on the immune recognition of viral E6 and E7 antigens. Twelve LCR/E6/E7 of the HPV-58 isolates were amplified and sequenced. A neighbor-joining phylogenetic tree was constructed by MEGA 7.0, followed by the secondary structure prediction of the related proteins using PSIPRED v3.3. The selection pressure acting on the HPV-58 E6 and E7 coding regions was estimated by Bayes empirical Bayes analysis of PAML 4.8. Meanwhile, the MHC class-I and II binding peptides were predicted by the ProPred-I server and ProPred server. The transcription factor binding sites in the HPV-58 LCR were analyzed using the JASPAR database. Twenty nine SNPs (20 in the LCR, 3 in the E6, 6 in the E7) were identified at 27 nucleotide sites across the HPV-58 LCR/E6/E7. From the most variable to the least variable, the nucleotide variations were LCR > E7 > E6. The combinations of all the SNPs resulted in 11 unique sequences, which were clustered into the A lineage (7 belong to A1, 2 belong to A2, and 2 belong to A3). An insertion (TGTCAGTTTCCT) was found between the nucleotide sites 7280 and 7281 in 2 variants, and a deletion (TTTAT) was found between 7429 and 7433 in 1 variant. The most common non-synonymous substitution V77A in the E7 was observed in the sequences encoding the α-helix. 63G in the E7 was determined to be the only one positively selected site in the HPV-58 E6/E7 sequences. Six non-synonymous amino acid substitutions (including S71F and K93 N in the E6, and T20I, G41R, G63S/D, and V77A in the E7) were affecting multiple putative epitopes for both CD4 + and CD8 + T-cells. In the LCR, C7265G and C7266T were the most variable sites and were the potential binding sites for the transcription factor SOX10. These results provide an insight into the intrinsic geographical relatedness and biological differences of the HPV-58 variants, and contribute to further research on the HPV-58 epidemiology, carcinogenesis, and therapeutic vaccine development.

[Central venous-arterial carbon dioxide tension to arterial-central venous oxygen content ratio combined with lactate clearance rate as early resuscitation goals of septic shock].

PubMed

Gao, X H; Li, P J; Cao, W

2018-02-13

Objective: To investigate the prognostic significance of central venous-arterial carbon dioxide tension to arterial-venous oxygen content ratio (Pcv-aCO(2)/Ca-cvO(2)) combined with lactate clearance rate (LCR) as early resuscitation goals of septic shock. Methods: One hundred and forty-five septic shock patients admitted to Second Department of Critical Care Medicine of Lanzhou University Second Hospital from March 2013 to May 2017 were enrolled in this study.All septic shock patients received an initial resuscitation therapy according to early goal-directed therapy.The arterial and central venous blood gases were measured simultaneously at baseline (T0) and 6 hours after resuscitation (T6). Pcv-aCO(2)/Ca-cvO(2) and LCR were calculated.Patients were classified into four groups according to Pcv-aCO(2)/Ca-cvO(2) and LCR at T6: group A, Pcv-aCO(2)/Ca-cvO(2)>1.8 and LCR<30%; group B, Pcv-aCO(2)/Ca-cvO(2)>1.8 and LCR≥30%; group C, Pcv-aCO(2)/Ca-cvO(2)≤1.8 and LCR<30%; group D, Pcv-aCO(2)/Ca-cvO(2)≤1.8 and LCR≥30%.General demographics, hemodynamic parameters, oxygen metabolism parameters, acute physiology and chronic health evaluation (APACHE Ⅱ) scores, sequential organ failure assessment (SOFA) scores, length of intensive care unit (ICU) stay, and 28-day mortality rate were compared among the 4 groups.A Kaplan-Meier curve showed the survival probabilities at day 28 using a log-rank test for multiple comparisons.Parameters were introduced into a Cox's proportional hazards regression model to analyze the prediction of 28-day mortality.Receiver operating characteristics (ROC) curves were constructed to evaluate the ability of Pcv-aCO(2)/Ca-cvO(2), LCR, Pcv-aCO(2)/Ca-cvO(2) combined with LCR at T6 to predict 28-day mortality. Results: Compared with patients in group A, patients from group D had the lower APACHE Ⅱ and SOFA score at day 3 ( t =-2.909, -3.630, both P <0.05), shorter ICU stay ( t =-2.575, P =0.011), and lower mortality rate at day 28 (χ(2)=3.124, P =0.011). Survival curves up to day 28, illustrated by Kaplan-Meier method, showed that group A had the shortest median survival time (χ(2)=10.332, P =0.016), difference between group A and group D was statistically significant (χ(2)=8.304, P =0.004). The Cox regression analysis revealed that Pcv-aCO(2)/Ca-cvO(2) ( RR =3.888, 95% CI : 2.443-6.189, P <0.001) and LCR ( RR =0.073, 95% CI : 0.008-0.640, P =0.018) at T6 were independent predictors of 28-day mortality.The area under ROC curve for Pcv-aCO(2)/Ca-cvO(2) combined with LCR (0.919, 95% CI : 0.862-0.958) was significantly greater than whether Pcv-aCO(2)/Ca-cvO(2) (0.862, 95% CI : 0.795-0.914) or LCR (0.820, 95% CI : 0.748-0.879) alone ( Z =2.032, 2.364, both P <0.05). Conclusion: Combination of Pcv-aCO(2)/Ca-cvO(2) and LCR is better than single parameter to predict the risk of adverse outcomes of septic shock patients, and may provide useful information for assessing the adequacy of resuscitation at early stage of septic shock.

[Comparison of the effect of fluid resuscitation as guided either by lactate clearance rate or by central venous oxygen saturation in patients with sepsis].

PubMed

Yu, Bin; Tian, Hui-yan; Hu, Zhen-jie; Zhao, Chai; Liu, Li-xia; Zhang, Yong; Zhu, Gui-jun; Wang, Lan-tao; Wu, Xin-hui; Li, Juan

2013-10-01

To compare the efficacy of fluid resuscitation as guided by lactate clearance rate (LCR) and central venous oxygen saturation (ScvO2) in patients with sepsis. A prospective randomized control study was conducted. Fifty patients diagnosed with severe sepsis or septic shock from January 2011 to February 2012 in department of critical care medicine of Fourth Hospital of Hebei Medical University were enrolled in the study. The patients were randomly divided into two groups according to the sequence (each n=25): ScvO2 group and LCR group. After ICU admission, the patients were treated symptomatically timely, and fluid resuscitation was started as early as possible according to Surviving Sepsis Campaign guidance for management of severe sepsis and septic shock 2008. Central venous pressure (CVP)≥8 mm Hg (1 mm Hg=0.133 kPa), mean arterial pressure (MAP)≥65 mm Hg and ScvO2≥0.70 served as goal values to accomplish the fluid resuscitation therapy in ScvO2 group, while CVP≥8 mm Hg, MAP≥65 mm Hg, LCR≥10% served as goal value to accomplish the fluid resuscitation therapy in LCR group. The general condition and clinical characteristics on arrival in ICU, changes in CVP, MAP, ScvO2, lactate level and/or LCR before (0 hour) and 3, 6, 72 hours after the start of fluid resuscitation and the other related conditions during the therapy were recorded. There was no significant difference in general data or clinical characteristics before the start of therapy, occurrence of organ dysfunction, or treatment measures during different time periods after start of fluid resuscitation. Compared with the condition immediately before fluid resuscitation, at 3 hours after start of fluid resuscitation, CVP were improved in LCR and ScvO2 groups (8.58±1.17 mm Hg vs. 6.33±1.21 mm Hg, 9.08±2.43 mm Hg vs. 5.33±0.98 mm Hg, both P<0.05); at 6 hours after start of fluid resuscitation, heart rate (HR) and respiratory rate (RR) were lowered in LCR and ScvO2 groups (HR: 96±18 bpm vs. 127±13 bpm, 98±13 bpm vs. 116±19 bpm, RR: 23±3 times/min vs. 33±9 times/min, 24±5 times/min vs. 35±6 times/min, all P<0.05), oxygenation index (PaO2/FiO2) was increased in LCR and ScvO2 groups (179±41 mm Hg vs. 86±21 mm Hg, 202±33 mm Hg vs. 95±17 mm Hg, both P<0.05), and there was no significant difference in MAP in both groups. There was no significant difference in all indexes between two groups. In LCR group, 3 hours after start of fluid resuscitation, lactate level was significantly decreased (2.81±0.18 mmol/L vs. 3.43±1.31 mmol/L, P<0.05). Compared with the value 3 hours after start of fluid resuscitation, LCR was significantly improved at 6 hours and 72 hours after start of fluid resuscitation in LCR group [(42.69±8.75)%, (48.87±9.69)% vs. (20.32±4.58)%, both P<0.05]. Compared with that immediately before fluid resuscitation, ScvO2 was significant improved in ScvO2 group at 3 hours after start of fluid resuscitation (0.65±0.04 vs. 0.53±0.06, P<0.05). There was no significant difference in success rate of fluid resuscitation comparing that of 6 hours and that of 72 hours [6 hours: 72% (18/25) vs. 64% (16/25), χ(2)=0.368, P=0.762; 72 hours: 88% (22/25) vs. 88% (22/25) ,χ(2)=0.000, P=1.000], length of ICU stay (8±3 days vs. 10±4 days, t=0.533, P=0.874), length of hospital stay (29±11 days vs. 35±16 days, t=0.692, P=0.531), improvement rate [84% (21/25) vs. 76%(19/25), χ(2)=0.500, P=0.480] or 28-day mortality [20% (5/25) vs. 28% (7/25), χ(2)=0.439, P=0.742] between LCR and ScvO2 groups. Both LCR and ScvO2 can be taken as the index in confirming the endpoint of fluid resuscitation for patients with severe sepsis and septic shock. Fluid resuscitation therapy under the guidance of LCR is accurate and reliable in patients with severe sepsis and septic shock.

Evaluation of a Rapid Assay for Detection of Chlamydia trachomatis Infections in Outpatient Clinics in South Kalimantan, Indonesia

PubMed Central

Widjaja, Susana; Cohen, Surekha; Brady, William E.; O'reilly, Kevin; Susanto; Wibowo, Ajar; Cahyono; Graham, Robert R.; Porter, Kevin R.

1999-01-01

A multicenter cross-sectional survey was conducted comparing a commercially available chlamydial optical immunoassay (OIA) to the chlamydial ligase chain reaction (LCR). Endocervical samples from 415 outpatients visiting clinics from three hospitals in South Kalimantan, Indonesia, were evaluated. Relative to the LCR, the overall sensitivity and specificity of the OIA were 31.6 and 98.9%, respectively. The sensitivity of the OIA varied among the three hospital laboratories, ranging from 20 to 50%. The OIA performance was slightly lower on samples from patients attending dermatovenereology clinics than on samples from nondermatovenereology clinic patients. The results indicate that the OIA did not perform well compared to LCR. PMID:10565960

Evaluation of a rapid assay for detection of Chlamydia trachomatis infections in outpatient clinics in South Kalimantan, Indonesia.

PubMed

Widjaja, S; Cohen, S; Brady, W E; O'reilly, K; Susanto; Wibowo, A; Cahyono; Graham, R R; Porter, K R

1999-12-01

A multicenter cross-sectional survey was conducted comparing a commercially available chlamydial optical immunoassay (OIA) to the chlamydial ligase chain reaction (LCR). Endocervical samples from 415 outpatients visiting clinics from three hospitals in South Kalimantan, Indonesia, were evaluated. Relative to the LCR, the overall sensitivity and specificity of the OIA were 31.6 and 98.9%, respectively. The sensitivity of the OIA varied among the three hospital laboratories, ranging from 20 to 50%. The OIA performance was slightly lower on samples from patients attending dermatovenereology clinics than on samples from nondermatovenereology clinic patients. The results indicate that the OIA did not perform well compared to LCR.

Energy transfer of nucleic acid products

NASA Astrophysics Data System (ADS)

Jung, Paul M.; Hu, Hsiang-Yun; Khalil, Omar S.

1995-04-01

Fluorescence energy transfer was investigated as a homogeneous detection method for the gapped ligase chain reaction (G-LCR). Oligonucleotides of a Chlamydia trachomatic G-LCR probe set were labeled with fluorescein as the donor and Texas Red as the acceptor fluorophore. Amplification and detection of 10 molecules of synthetic target was demonstrated in spiked urine samples.

Facilitating Career Development through Super's Life Career Rainbow.

ERIC Educational Resources Information Center

Okocha, Aneneosa A.

Super's life-span life-space theory offers a developmental framework for career counseling. This paper provides a brief overview of Super's theory of Life Career Rainbow (LCR) segment. The LCR feature is useful for identifying the stage of a client's career development and in formulating goals for counseling. The assessment is accomplished by…

77 FR 27186 - Endangered and Threatened Species; Take of Anadromous Fish

Federal Register 2010, 2011, 2012, 2013, 2014

2012-05-09

...) is seeking a 5- year permit to take juvenile LCR coho salmon and steelhead during research designed... juvenile LCR coho salmon and steelhead during research designed to assess the distribution of bull trout in... steelhead during the course of research designed to provide information on the dynamics and use of cold...

Nested Inversion Polymorphisms Predispose Chromosome 22q11.2 to Meiotic Rearrangements.

PubMed

Demaerel, Wolfram; Hestand, Matthew S; Vergaelen, Elfi; Swillen, Ann; López-Sánchez, Marcos; Pérez-Jurado, Luis A; McDonald-McGinn, Donna M; Zackai, Elaine; Emanuel, Beverly S; Morrow, Bernice E; Breckpot, Jeroen; Devriendt, Koenraad; Vermeesch, Joris R

2017-10-05

Inversion polymorphisms between low-copy repeats (LCRs) might predispose chromosomes to meiotic non-allelic homologous recombination (NAHR) events and thus lead to genomic disorders. However, for the 22q11.2 deletion syndrome (22q11.2DS), the most common genomic disorder, no such inversions have been uncovered as of yet. Using fiber-FISH, we demonstrate that parents transmitting the de novo 3 Mb LCR22A-D 22q11.2 deletion, the reciprocal duplication, and the smaller 1.5 Mb LCR22A-B 22q11.2 deletion carry inversions of LCR22B-D or LCR22C-D. Hence, the inversions predispose chromosome 22q11.2 to meiotic rearrangements and increase the individual risk for transmitting rearrangements. Interestingly, the inversions are nested or flanking rather than coinciding with the deletion or duplication sizes. This finding raises the possibility that inversions are a prerequisite not only for 22q11.2 rearrangements but also for all NAHR-mediated genomic disorders. Copyright © 2017. Published by Elsevier Inc.

Race, deprivation, and immigrant isolation: The spatial demography of air-toxic clusters in the continental United States.

PubMed

Liévanos, Raoul S

2015-11-01

This article contributes to environmental inequality outcomes research on the spatial and demographic factors associated with cumulative air-toxic health risks at multiple geographic scales across the United States. It employs a rigorous spatial cluster analysis of census tract-level 2005 estimated lifetime cancer risk (LCR) of ambient air-toxic emissions from stationary (e.g., facility) and mobile (e.g., vehicular) sources to locate spatial clusters of air-toxic LCR risk in the continental United States. It then tests intersectional environmental inequality hypotheses on the predictors of tract presence in air-toxic LCR clusters with tract-level principal component factor measures of economic deprivation by race and immigrant status. Logistic regression analyses show that net of controls, isolated Latino immigrant-economic deprivation is the strongest positive demographic predictor of tract presence in air-toxic LCR clusters, followed by black-economic deprivation and isolated Asian/Pacific Islander immigrant-economic deprivation. Findings suggest scholarly and practical implications for future research, advocacy, and policy. Copyright © 2015 Elsevier Inc. All rights reserved.

Neutralization of Yersinia pestis-mediated macrophage cytotoxicity by anti-LcrV antibodies and its correlation with protective immunity in a mouse model of bubonic plague.

PubMed

Zauberman, Ayelet; Cohen, Sara; Levy, Yinon; Halperin, Gideon; Lazar, Shirley; Velan, Baruch; Shafferman, Avigdor; Flashner, Yehuda; Mamroud, Emanuelle

2008-03-20

Plague is a life-threatening disease caused by Yersinia pestis, for which effective-licensed vaccines and reliable predictors of in vivo immunity are lacking. V antigen (LcrV) is a major Y. pestis virulence factor that mediates translocation of the cytotoxic Yersinia protein effectors (Yops). It is a well-established protective antigen and a part of currently tested plague subunit vaccines. We have developed a highly sensitive in vitro macrophage cytotoxicity neutralization assay which is mediated by anti-LcrV antibodies; and studied the potential use of these neutralizing antibodies as an in vitro correlate of plague immunity in mice. The assay is based on a Y. pestis strain with enhanced cytotoxicity to macrophages in which endogenous yopJ was replaced by the more effectively translocated yopP of Y. enterocolitica O:8. Mice passively immunized with rabbit anti-LcrV IgG or actively immunized with recombinant LcrV were protected against lethal doses of a virulent Y. pestis strain, in a mouse model of bubonic plague. This protection significantly correlated with the in vitro neutralizing activity of the antisera but not with their corresponding ELISA titers. In actively immunized mice, a cutoff value for serum neutralizing activity, above which survival was assured with high degree of confidence, could be established for different vaccination regimes. The impact of overall findings on the potential use of serum neutralizing activity as a correlate of protective immunity is discussed.

The hematopoietic regulator TAL1 is required for chromatin looping between the β-globin LCR and human γ-globin genes to activate transcription

PubMed Central

Yun, Won Ju; Kim, Yea Woon; Kang, Yujin; Lee, Jungbae; Dean, Ann; Kim, AeRi

2014-01-01

TAL1 is a key hematopoietic transcription factor that binds to regulatory regions of a large cohort of erythroid genes as part of a complex with GATA-1, LMO2 and Ldb1. The complex mediates long-range interaction between the β-globin locus control region (LCR) and active globin genes, and although TAL1 is one of the two DNA-binding complex members, its role is unclear. To explore the role of TAL1 in transcription activation of the human γ-globin genes, we reduced the expression of TAL1 in erythroid K562 cells using lentiviral short hairpin RNA, compromising its association in the β-globin locus. In the TAL1 knockdown cells, the γ-globin transcription was reduced to 35% and chromatin looping of the Gγ-globin gene with the LCR was disrupted with decreased occupancy of the complex member Ldb1 and LMO2 in the locus. However, GATA-1 binding, DNase I hypersensitive site formation and several histone modifications were largely maintained across the β-globin locus. In addition, overexpression of TAL1 increased the γ-globin transcription and increased interaction frequency between the Gγ-globin gene and LCR. These results indicate that TAL1 plays a critical role in chromatin loop formation between the γ-globin genes and LCR, which is a critical step for the transcription of the γ-globin genes. PMID:24470145

LDB1-mediated enhancer looping can be established independent of mediator and cohesin.

PubMed

Krivega, Ivan; Dean, Ann

2017-08-21

Mechanistic studies in erythroid cells indicate that LDB1, as part of a GATA1/TAL1/LMO2 complex, brings erythroid-expressed genes into proximity with enhancers for transcription activation. The role of co-activators in establishing this long-range interaction is poorly understood. Here we tested the contributions of the RNA Pol II pre-initiation complex (PIC), mediator and cohesin to establishment of locus control region (LCR)/β-globin proximity. CRISPR/Cas9 editing of the β-globin promoter to eliminate the RNA Pol II PIC by deleting the TATA-box resulted in loss of transcription, but enhancer-promoter interaction was unaffected. Additional deletion of the promoter GATA1 site eliminated LDB1 complex and mediator occupancy and resulted in loss of LCR/β-globin proximity. To separate the roles of LDB1 and mediator in LCR looping, we expressed a looping-competent but transcription-activation deficient form of LDB1 in LDB1 knock down cells: LCR/β-globin proximity was restored without mediator core occupancy. Further, Cas9-directed tethering of mutant LDB1 to the β-globin promoter forced LCR loop formation in the absence of mediator or cohesin occupancy. Moreover, ENCODE data and our chromatin immunoprecipitation results indicate that cohesin is almost completely absent from validated and predicted LDB1-regulated erythroid enhancer-gene pairs. Thus, lineage specific factors largely mediate enhancer-promoter looping in erythroid cells independent of mediator and cohesin. Published by Oxford University Press on behalf of Nucleic Acids Research 2017.

Selection for intrinsic endurance modifies endocrine stress responsiveness

PubMed Central

Waters, R Parrish; Renner, Kenneth J; Summers, Cliff H; Watt, Michael L; Forster, Gina L; Koch, Lauren G; Britton, Steven L; Swallow, John G

2010-01-01

Physical exercise dampens an individual’s stress response and decreases symptoms of anxiety and depression disorders. While the extrinsic relationship of exercise and psychological state are established, their intrinsic relationship is unresolved. We investigated the potential intrinsic relationship of exercise with stress responsiveness using NIH rats bidirectionally selected for intrinsic endurance capacity. Selection resulted in two populations, one with high intrinsic endurance (high capacity runners; HCR) and one with low intrinsic endurance (low capacity runners; LCR). Animals from these populations were subjected to the elevated plus maze (EPM) and novel environment to assess levels of anxiety-like behavior, and to restraint stress to determine stress responsiveness. Pre-test plasma corticosterone levels and the response of plasma corticosterone to exposure to the EPM and restraint were analyzed using ELISA. A dexamethasone suppression test was performed to assess negative feedback tone of corticosterone release. Pre-test plasma corticosterone levels were similar between LCR and HCR, and these populations had similar behavioral and corticosterone responses to the EPM. Following restraint, HCR animals exhibited more anxiotypic behavior than LCR animals on the EPM, and exhibited an increase in plasma corticosterone following EPM and restraint that was not observed in LCR animals. HCR animals also exhibited more anxiotypic behavior in the novel environment compared to LCR animals. Plasma corticosterone levels were equally reduced in both populations following dexamethasone administration. Overall, our data suggest a positive genetic relationship between exercise endurance and stress responsiveness, which is at odds with the established extrinsic relationship of these traits. PMID:20682296

The hematopoietic regulator TAL1 is required for chromatin looping between the β-globin LCR and human γ-globin genes to activate transcription.

PubMed

Yun, Won Ju; Kim, Yea Woon; Kang, Yujin; Lee, Jungbae; Dean, Ann; Kim, AeRi

2014-04-01

TAL1 is a key hematopoietic transcription factor that binds to regulatory regions of a large cohort of erythroid genes as part of a complex with GATA-1, LMO2 and Ldb1. The complex mediates long-range interaction between the β-globin locus control region (LCR) and active globin genes, and although TAL1 is one of the two DNA-binding complex members, its role is unclear. To explore the role of TAL1 in transcription activation of the human γ-globin genes, we reduced the expression of TAL1 in erythroid K562 cells using lentiviral short hairpin RNA, compromising its association in the β-globin locus. In the TAL1 knockdown cells, the γ-globin transcription was reduced to 35% and chromatin looping of the (G)γ-globin gene with the LCR was disrupted with decreased occupancy of the complex member Ldb1 and LMO2 in the locus. However, GATA-1 binding, DNase I hypersensitive site formation and several histone modifications were largely maintained across the β-globin locus. In addition, overexpression of TAL1 increased the γ-globin transcription and increased interaction frequency between the (G)γ-globin gene and LCR. These results indicate that TAL1 plays a critical role in chromatin loop formation between the γ-globin genes and LCR, which is a critical step for the transcription of the γ-globin genes.

Involvement of CD8+ T cell-mediated immune responses in LcrV DNA vaccine induced protection against lethal Yersinia pestis challenge.

PubMed

Wang, Shixia; Goguen, Jon D; Li, Fusheng; Lu, Shan

2011-09-09

Yersinia pestis (Y. pestis) is the causative pathogen of plague, a highly fatal disease for which an effective vaccine, especially against mucosal transmission, is still not available. Like many bacterial infections, antigen-specific antibody responses have been traditionally considered critical, if not solely responsible, for vaccine-induced protection against Y. pestis. Studies in recent years have suggested the importance of T cell immune responses against Y. pestis infection but information is still limited about the details of Y. pestis antigen-specific T cell immune responses. In current report, studies are conducted to identify the presence of CD8+ T cell epitopes in LcrV protein, the leading antigen of plague vaccine development. Furthermore, depletion of CD8+ T cells in LcrV DNA vaccinated Balb/C mice led to reduced protection against lethal intranasal challenge of Y. pestis. These findings establish that an LcrV DNA vaccine is able to elicit CD8+ T cell immune responses against specific epitopes of this key plague antigen and that a CD8+ T cell immune response is involved in LcrV DNA vaccine-elicited protection. Future studies in plague vaccine development will need to examine if the presence of detectable T cell immune responses, in particular CD8+ T-cell immune responses, will enhance the protection against Y. pestis in higher animal species or humans. Copyright © 2010 Elsevier Ltd. All rights reserved.

Effects of intrinsic aerobic capacity and ovariectomy on voluntary wheel running and nucleus accumbens dopamine receptor gene expression

PubMed Central

Park, Young-Min; Kanaley, Jill A.; Padilla, Jaume; Zidon, Terese; Welly, Rebecca J.; Will, Matthew J.; Britton, Steven L.; Koch, Lauren G.; Ruegsegger, Gregory N.; Booth, Frank W.; Thyfault, John P.; Vieira-Potter, Victoria J.

2016-01-01

Rats selectively bred for high (HCR) and low (LCR) aerobic capacity show a stark divergence in wheel running behavior, which may be associated with dopamine (DA) system in the brain. HCR possess greater motivation for voluntary running along with greater brain DA activity compared to LCR. We recently demonstrated that HCR are not immune to ovariectomy (OVX)-associated reductions in spontaneous cage (i.e. locomotor) activity. Whether HCR and LCR rats differ in their OVX-mediated voluntary wheel running response is unknown. PURPOSE To determine whether HCR are protected from OVX-associated reduction in voluntary wheel running. METHODS Forty female HCR and LCR rats (age ~27 weeks) had either SHM or OVX operations, and given access to a running wheel for 11 weeks. Weekly wheel running distance was monitored throughout the intervention. Nucleus accumbens (NAc) was assessed for mRNA expression of DA receptors at sacrifice. RESULTS Compared to LCR, HCR ran greater distance and had greater ratio of excitatory/inhibitory DA mRNA expression (both line main effects, P<0.05). Wheel running distance was significantly, positively correlated with the ratio of excitatory/inhibitory DA mRNA expression across animals. In both lines, OVX reduced wheel running (P<0.05). Unexpectedly, although HCR started with significantly greater voluntary wheel running, they had greater OVX-induced reduction in wheel running than LCR such that no differences were found 11 weeks after OVX between HCROVX and LCROVX (interaction, P<0.05). This significant reduction in wheel running in HCR was associated with an OVX-mediated reduction in the ratio of excitatory/inhibitory DA mRNA expression. CONCLUSION DA system in the NAc region may play a significant role in motivation to run in female rats. Compared to LCR, HCR rats run significantly more, which associates with greater ratio of excitatory/inhibitory DA mRNA expression. However, despite greater inherent motivation to run and an associated brain DA mRNA expression profile, these HCR rats are not protected against OVX-induced reduction in wheel running. The impairment in wheel running in HCR rats may be partially explained by their reduced ratio of excitatory/inhibitory DA receptor mRNA expression. PMID:27297873

Effects of intrinsic aerobic capacity and ovariectomy on voluntary wheel running and nucleus accumbens dopamine receptor gene expression.

PubMed

Park, Young-Min; Kanaley, Jill A; Padilla, Jaume; Zidon, Terese; Welly, Rebecca J; Will, Matthew J; Britton, Steven L; Koch, Lauren G; Ruegsegger, Gregory N; Booth, Frank W; Thyfault, John P; Vieira-Potter, Victoria J

2016-10-01

Rats selectively bred for high (HCR) and low (LCR) aerobic capacity show a stark divergence in wheel running behavior, which may be associated with the dopamine (DA) system in the brain. HCR possess greater motivation for voluntary running along with greater brain DA activity compared to LCR. We recently demonstrated that HCR are not immune to ovariectomy (OVX)-associated reductions in spontaneous cage (i.e. locomotor) activity. Whether HCR and LCR rats differ in their OVX-mediated voluntary wheel running response is unknown. To determine whether HCR are protected from OVX-associated reduction in voluntary wheel running. Forty female HCR and LCR rats (age ~27weeks) had either SHM or OVX operations, and given access to a running wheel for 11weeks. Weekly wheel running distance was monitored throughout the intervention. Nucleus accumbens (NAc) was assessed for mRNA expression of DA receptors at sacrifice. Compared to LCR, HCR ran greater distance and had greater ratio of excitatory/inhibitory DA mRNA expression (both line main effects, P<0.05). Wheel running distance was significantly, positively correlated with the ratio of excitatory/inhibitory DA mRNA expression across animals. In both lines, OVX reduced wheel running (P<0.05). Unexpectedly, although HCR started with significantly greater voluntary wheel running, they had greater OVX-induced reduction in wheel running than LCR such that no differences were found 11weeks after OVX between HCROVX and LCROVX (interaction, P<0.05). This significant reduction in wheel running in HCR was associated with an OVX-mediated reduction in the ratio of excitatory/inhibitory DA mRNA expression. The DA system in the NAc region may play a significant role in motivation to run in female rats. Compared to LCR, HCR rats run significantly more, which associates with greater ratio of excitatory/inhibitory DA mRNA expression. However, despite greater inherent motivation to run and an associated brain DA mRNA expression profile, HCR rats are not protected against OVX-induced reduction in wheel running or OVX-mediated reduction in the ratio of excitatory/inhibitory DA receptor mRNA expression. OVX-mediated reduction in motivated physical activity may be partially explained by a reduced ratio of excitatory/inhibitory DA receptor mRNA expression for which intrinsic fitness does not confer protection. Copyright © 2016 Elsevier Inc. All rights reserved.

Autogenic incision and terrace formation resulting from abrupt late-glacial base-level fall, lower Chippewa River, Wisconsin, USA

NASA Astrophysics Data System (ADS)

Faulkner, Douglas J.; Larson, Phillip H.; Jol, Harry M.; Running, Garry L.; Loope, Henry M.; Goble, Ronald J.

2016-08-01

A paucity of research exists regarding the millennial-scale response of inland alluvial streams to abrupt base-level fall. Studies of modern systems indicate that, over short time scales, the response is a diffusion-like process of upstream-propagating incision. In contrast, evidence from the lower Chippewa River (LCR), located in the upper Midwest of the USA, suggests that autogenic controls operating over time scales of several millennia can overwhelm diffusion, resulting in incision that is prolonged and episodic. During the Last Glacial Maximum, the LCR drained the Chippewa Lobe of the Laurentide Ice Sheet to the glacial upper Mississippi River (UMR). As a meltwater stream, it aggraded and filled its valley with glacial outwash, as did its largest tributaries, which were also meltwater streams. Its nonglacial tributaries aggraded, too, filling their valleys with locally derived sediment. During deglaciation, the UMR incised at least twice, abruptly lowering the LCR's base level - 15 m at 16 ka or earlier and an additional 40 m at ca. 13.4 ka. Each of these base-level falls initiated incision of the LCR, led by upstream migrating knickpoints. The propagation of incision has, however, been a lengthy process. The optically stimulated luminescence (OSL) ages of terrace alluvium indicate that, by 13.5 ka, incision had advanced up the LCR only 15 km, and by 9 ka, only 55 km. The process has also been episodic, resulting in the formation of fill-cut terraces (inferred from GPR surveys and exposures of terrace alluvium) that are younger and more numerous in the upstream direction. Autogenic increases in sediment load and autogenic bed armoring, the result of periodic tributary-stream rejuvenation and preferential winnowing of fines by the incising river, may have periodically caused knickpoint migration and incision to slow and possibly stop, allowing lateral erosion and floodplain formation to dominate. A decline in sediment flux from stabilizing incised tributary stream systems would have led to renewed knickpoint migration and incision when floods of sufficient magnitude to breach the channel armor occurred. Minimal floodplain development along the upper section of the present-day LCR, along with the channel morphology of an unstable wandering gravel-bed river immediately downstream from it, suggest that the river is still responding to the base-level falls that happened many millennia ago. The autogenic controls on the LCR's response to UMR incision are a direct consequence of the thick fills of noncohesive sediment that accumulated in its valley and the valleys of its tributary streams during the Late Wisconsinan, making the LCR a prime example of a former proglacial river that remains a paraglacial fluvial system.

Biology and Molecular Characterization of Cucurbit leaf crumple virus, an Emergent Cucurbit-Infecting Begomovirus in the Imperial Valley of California

USDA-ARS?s Scientific Manuscript database

Cucurbit leaf crumple virus (CuLCrV) is an emergent and potentially economically important bipartite begomovirus first identified in volunteer watermelon plants in the Imperial Valley of southern California in 1998. Field surveys indicated that CuLCrV has become established in the Imperial Valley; a...

Epidemiological Analysis of Multi-Virus Infections of Watermelon in Experimental Fields in Southwest Florida

USDA-ARS?s Scientific Manuscript database

The whitefly-transmitted viruses Squash vein yellowing virus (SqVYV) and Cucurbit leaf crumple virus (CuLCrV) have had serious impact on watermelon production in west-central and southwest Florida in recent years. We monitored the progress of SqVYV and CuLCrV and whitefly density in 2.5 acre experim...

Defective GATA-3 expression in Th2 LCR-deficient mice.

PubMed

Hwang, Soo Seok; Kim, Kiwan; Lee, Gap Ryol

2011-07-15

Th2 cell differentiation is critically influenced by transcription factor GATA-3 and by various cis-acting elements including enhancers, silencers and a locus control region (LCR) in the Th2 cytokine locus. Th2 LCR-deficient Th2 cells completely lost the expression of GATA-3 and the phosphorylation of STAT6. Histone 3 lysine 4 (H3-K4) was hypomethylated in the gata3 locus in these cells. GATA-3 and STAT6 bound several regulatory regions in the gata3 locus and transactivated the expression of the gata3 gene. These results suggest that Th2 differentiation program stimulates feed-forward regulation of gata3 gene expression. Copyright © 2011 Elsevier Inc. All rights reserved.

Delocalized periodic vibrations in nonlinear LC and LCR electrical chains

NASA Astrophysics Data System (ADS)

Chechin, G. M.; Shcherbinin, S. A.

2015-05-01

We consider electrical LC- and LCR-chains consisting of N cells. In the LC-chain each cell contains a linear inductor L and a nonlinear capacitor C, while the cell in the LCR-chain include additionally a resistor R and an voltage source. It is assumed that voltage dependence of capacitors represents an even function. Such capacitors have implemented by some experimental groups studying propagation of electrical signals in the lines constructed on MOS and CMOS substrates. In these chains, we study dynamical regimes representing nonlinear normal modes (NNMs) by Rosenberg. We prove that maximum possible number of symmetry-determined NNMs which can be excited in the considered chains is equal to 5. The stability of these modes for different N is studied with the aid of the group-theoretical method [Physical Review E 73 (2006) 36216] which allows to simplify radically the variational systems appearing in the Floquet stability analysis. For NNMs in LC-chain, the scaling of the voltage stability threshold in the thermodynamic limit (N → ∞) is determined. It is shown that the above group theoretical method can be also used for studying stability of NNMs in the LCR-chains.

Factors controlling the abundance of rainbow trout in the Colorado River in Grand Canyon in a reach utilized by endangered humpback chub

USGS Publications Warehouse

Korman, Josh; Yard, Michael D.; Yackulic, Charles B.

2015-01-01

We estimated the abundance, survival, movement, and recruitment of non-native rainbow trout in the Colorado River in Grand Canyon to determine what controls their abundance near the Little Colorado River (LCR) confluence where endangered humpback chub rear. Over a 3-year period, we tagged more than 70,000 trout and recovered over 8,200 tagged fish. Trout density was highest (10,000-25,000 fish/km) in the reach closest to Glen Canyon Dam where the majority of trout recruitment occurs, and was 30-50-fold lower (200-800 fish/km) in reaches near the LCR confluence ~100 km downstream. The extent of rainbow trout movement was limited with less than 1% of recaptures making movements greater than 20 km. However, due to high trout densities in upstream source areas, this small dispersal rate was sufficient to explain the 3-fold increase in the relatively small population near the LCR. Reducing dispersal rates of trout from upstream sources is the most feasible solution to maintain low densities near the LCR to minimize negative effects of competition and predation on humpback chub.

Sequence change in the HS2-LCR and Ggamma-globin gene promoter region of sickle cell anemia patients.

PubMed

Adorno, E V; Moura-Neto, J P; Lyra, I; Zanette, A; Santos, L F O; Seixas, M O; Reis, M G; Goncalves, M S

2008-02-01

The fetal hemoglobin (HbF) levels and betaS-globin gene haplotypes of 125 sickle cell anemia patients from Brazil were investigated. We sequenced the Ggamma- and Agamma-globin gene promoters and the DNase I-2 hypersensitive sites in the locus control regions (HS2-LCR) of patients with HbF level disparities as compared to their betaS haplotypes. Sixty-four (51.2%) patients had CAR/Ben genotype; 36 (28.8%) Ben/Ben; 18 (14.4%) CAR/CAR; 2 (1.6%) CAR/Atypical; 2 (1.6%) Ben/Cam; 1 (0.8%) CAR/Cam; 1 (0.8%) CAR/Arab-Indian, and 1 (0.8%) Sen/Atypical. The HS2-LCR sequence analyses demonstrated a c.-10.677G>A change in patients with the Ben haplotype and high HbF levels. The Gg gene promoter sequence analyses showed a c.-157T>C substitution shared by all patients, and a c.-222_-225del related to the Cam haplotype. These results identify new polymorphisms in the HS2-LCR and Gg-globin gene promoter. Further studies are required to determine the correlation between HbF synthesis and the clinical profile of sickle cell anemia patients.

Interphase FISH screening for the LCR-mediated common rearrangement of isochromosome 17q in primary myelofibrosis.

PubMed

Bien-Willner, Gabriel A; Stankiewicz, Paweł; Lupski, James R; Northup, Jill K; Velagaleti, Gopalrao V N

2005-08-01

Non-allelic homologous recombination (NAHR) between low-copy repeats (LCRs) has been implicated recently in somatic rearrangements including isochromosome i(17q), which is associated with hematologic malignancies as well as solid tumors. In hematological malignancies, the most common i(17q) breakpoint results from LCR-mediated NAHR, which creates a dicentric chromosome, idic(17)(p11.2). We report an elderly patient who presented with primary myelofibrosis (MF) with myeloid metaplasia (MMM), associated with idic(17)(p11.2) as the sole chromosomal abnormality, making this the first idic(17)(p11.2) myeloproliferative case reported in which the breakpoints are mapped to the breakpoint cluster region in proximal 17p. The rearrangement breakpoint maps to the previously defined LCR cluster, further suggesting that the genomic architecture of proximal 17p may be responsible for the formation of the majority of i(17q) cases. We describe our development of a rapid screening test using interphase FISH to detect idic(17)(p11.2), discuss the potential prognostic value of this molecular diagnostic test, and examine the relevance of LCR-mediated NAHR to common rearrangements in neoplasms. Copyright (c) 2005 Wiley-Liss, Inc.

The C-terminal domain of Nrf1 negatively regulates the full-length CNC-bZIP factor and its shorter isoform LCR-F1/Nrf1β; both are also inhibited by the small dominant-negative Nrf1γ/δ isoforms that down-regulate ARE-battery gene expression.

PubMed

Zhang, Yiguo; Qiu, Lu; Li, Shaojun; Xiang, Yuancai; Chen, Jiayu; Ren, Yonggang

2014-01-01

The C-terminal domain (CTD, aa 686-741) of nuclear factor-erythroid 2 p45-related factor 1 (Nrf1) shares 53% amino acid sequence identity with the equivalent Neh3 domain of Nrf2, a homologous transcription factor. The Neh3 positively regulates Nrf2, but whether the Neh3-like (Neh3L) CTD of Nrf1 has a similar role in regulating Nrf1-target gene expression is unknown. Herein, we report that CTD negatively regulates the full-length Nrf1 (i.e. 120-kDa glycoprotein and 95-kDa deglycoprotein) and its shorter isoform LCR-F1/Nrf1β (55-kDa). Attachment of its CTD-adjoining 112-aa to the C-terminus of Nrf2 yields the chimaeric Nrf2-C112Nrf1 factor with a markedly decreased activity. Live-cell imaging of GFP-CTD reveals that the extra-nuclear portion of the fusion protein is allowed to associate with the endoplasmic reticulum (ER) membrane through the amphipathic Neh3L region of Nrf1 and its basic c-tail. Thus removal of either the entire CTD or the essential Neh3L portion within CTD from Nrf1, LCR-F1/Nrf1β and Nrf2-C112Nrf1, results in an increase in their transcriptional ability to regulate antioxidant response element (ARE)-driven reporter genes. Further examinations unravel that two smaller isoforms, 36-kDa Nrf1γ and 25-kDa Nrf1δ, act as dominant-negative inhibitors to compete against Nrf1, LCR-F1/Nrf1β and Nrf2. Relative to Nrf1, LCR-F1/Nrf1β is a weak activator, that is positively regulated by its Asn/Ser/Thr-rich (NST) domain and acidic domain 2 (AD2). Like AD1 of Nrf1, both AD2 and NST domain of LCR-F1/Nrf1β fused within two different chimaeric contexts to yield Gal4D:Nrf1β607 and Nrf1β:C270Nrf2, positively regulate their transactivation activity of cognate Gal4- and Nrf2-target reporter genes. More importantly, differential expression of endogenous ARE-battery genes is attributable to up-regulation by Nrf1 and LCR-F1/Nrf1β and down-regulation by Nrf1γ and Nrf1δ.

EVALUATING THE IMPACT OF WATER CHEMISTRY ON CUPROSOLVENCY AND COPPER CORROSION BY-PRODUCT USING A SIMPLE COPPER PIPE RECIRCULATING LOOP SYSTEM

EPA Science Inventory

1991, EPA publicized the Lead and Copper Rule (LCR),which set regulations to minimize the amount of lead copper in drinking water. The LCR set the copper action level at 1.3 mg/L in more then 10% of customer’s first-draw taps sampled. Potential health effects of copper include vo...

Using the Arduino with MakerPlot software for the display of resonance curves characterisic of a series LCR circuit

NASA Astrophysics Data System (ADS)

Atkin, Keith

2016-11-01

This paper shows how very simple circuitry attached to an Arduino microcontroller can be used for the measurement of both frequency and amplitude of a sinusoidal signal. It is also shown how the addition of a readily available software package, MakerPlot, can facilitate the display and investigation of resonance curves for a series LCR circuit.

Divergent skeletal muscle respiratory capacities in rats artificially selected for high and low running ability: a role for Nor1?

PubMed Central

Stephenson, Erin J.; Stepto, Nigel K.; Koch, Lauren G.; Britton, Steven L.

2012-01-01

Inactivity-related diseases are becoming a huge burden on Western society. While there is a major environmental contribution to metabolic health, the intrinsic properties that predispose or protect against particular health traits are harder to define. We used rat models of inborn high running capacity (HCR) and low running capacity (LCR) to determine inherent differences in mitochondrial volume and function, hypothesizing that HCR rats would have greater skeletal muscle respiratory capacity due to an increase in mitochondrial number. Additionally, we sought to determine if there was a link between the expression of the orphan nuclear receptor neuron-derived orphan receptor (Nor)1, a regulator of oxidative metabolism, and inherent skeletal muscle respiratory capacity. LCR rats were 28% heavier (P < 0.0001), and fasting serum insulin concentrations were 62% greater than in HCR rats (P = 0.02). In contrast, HCR rats had better glucose tolerance and reduced adiposity. In the primarily oxidative soleus muscle, maximal respiratory capacity was 21% greater in HCR rats (P = 0.001), for which the relative contribution of fat oxidation was 20% higher than in LCR rats (P = 0.02). This was associated with increased citrate synthase (CS; 33%, P = 0.009) and β-hydroxyacyl-CoA (β-HAD; 33%, P = 0.0003) activities. In the primarily glycolytic extensor digitum longus muscle, CS activity was 29% greater (P = 0.01) and β-HAD activity was 41% (P = 0.0004) greater in HCR rats compared with LCR rats. Mitochondrial DNA copy numbers were also elevated in the extensor digitum longus muscles of HCR rats (35%, P = 0.049) and in soleus muscles (44%, P = 0.16). Additionally, HCR rats had increased protein expression of individual mitochondrial respiratory complexes, CS, and uncoupling protein 3 in both muscle types (all P < 0.05). In both muscles, Nor1 protein was greater in HCR rats compared with LCR rats (P < 0.05). We propose that the differential expression of Nor1 may contribute to the differences in metabolic regulation between LCR and HCR phenotypes. PMID:22936731

Exercise Prevents Enhanced Postoperative Neuroinflammation and Cognitive Decline and Rectifies the Gut Microbiome in a Rat Model of Metabolic Syndrome.

PubMed

Feng, Xiaomei; Uchida, Yosuke; Koch, Lauren; Britton, Steve; Hu, Jun; Lutrin, David; Maze, Mervyn

2017-01-01

Postoperative cognitive decline (PCD) can affect in excess of 10% of surgical patients and can be considerably higher with risk factors including advanced age, perioperative infection, and metabolic conditions such as obesity and insulin resistance. To define underlying pathophysiologic processes, we used animal models including a rat model of metabolic syndrome generated by breeding for a trait of low aerobic exercise tolerance. After 35 generations, the low capacity runner (LCR) rats differ 10-fold in their aerobic exercise capacity from high capacity runner (HCR) rats. The LCR rats respond to surgical procedure with an abnormal phenotype consisting of exaggerated and persistent PCD and failure to resolve neuroinflammation. We determined whether preoperative exercise can rectify the abnormal surgical phenotype. Following institutional approval of the protocol each of male LCR and male HCR rats were randomly assigned to four groups and subjected to isoflurane anesthesia and tibia fracture with internal fixation (surgery) or anesthesia alone (sham surgery) and to a preoperative exercise regimen that involved walking for 10 km on a treadmill over 6 weeks (exercise) or being placed on a stationary treadmill (no exercise). Feces were collected before and after exercise for assessment of gut microbiome. Three days following surgery or sham surgery the rats were tested for ability to recall a contextual aversive stimulus in a trace fear conditioning paradigm. Thereafter some rats were euthanized and the hippocampus harvested for analysis of inflammatory mediators. At 3 months, the remainder of the rats were tested for memory recall by the probe test in a Morris Water Maze. Postoperatively, LCR rats exhibited exaggerated cognitive decline both at 3 days and at 3 months that was prevented by preoperative exercise. Similarly, LCR rats had excessive postoperative neuroinflammation that was normalized by preoperative exercise. Diversity of the gut microbiome in the LCR rats improved after exercise. Preoperative exercise eliminated the metabolic syndrome risk for the abnormal surgical phenotype and was associated with a more diverse gut microbiome. Prehabilitation with exercise should be considered as a possible intervention to prevent exaggerated and persistent PCD in high-risk settings.

Virulence markers of LCR plasmid in Indian isolates of Yersinia pestis.

PubMed

Khushiramani, Rekha; Tuteja, Urmil; Shukla, Jyoti; Panikkar, Anupama; Batra, Harsh Vardhan

2006-01-01

Presence of 10 important yop genes in Yersinia pestis isolates (18 in number) of Indian origin from 1994 plague outbreak regions of Maharashtra (6 Rattus rattus & Tetera indica rodents) and Gujarat (11 from human patients, 1 from R. rattus) and from plague endemic regions of the Deccan plateau (8 from T. indica) was located by PCR and specific enzyme immunoassay. PCRs were standardized for six effector yops (YopE, YopH, YopJ, YopM, YopO and YopT), three translocator yops (YopB, YopD and YopK) and a regulator LcrV gene. Amplification of all the 10 yop genes was observed in isolates recovered from pneumonic patients and in 5 of 7 rodents from outbreak regions. Among these, amplification of the yopD gene was absent in all eight isolates, and that of yopM in all except one (10R). One of the isolates from rodents of the Deccan plateau (24H) was consistently negative for all the yops. Cloning and expression of truncated yopM (780 bp), yopB (700 bp) and lcrV (796 bp) genes in pQE vectors with SG13009 host cells yielded recombinant proteins for generation of monoclonal antibodies for further use in enzyme immunoassay. Ten stable reactive clones for YopB, nine for YopM and six for LcrV were obtained, all of them exhibiting specific reactions only to Y. pestis. Testing of 26 Y. pestis isolates by monoclonal antibody dot-ELISA and Western blotting provided results identical to PCR, suggesting that the isolates that failed to show PCR amplification also had no expression of their respective proteins. The Y. pestis isolates of outbreak regions had their virulence factors intact in the LCR plasmid. Yersinia pestis isolates recovered from rodents of the Deccan plateau were relatively heterogeneous. It appears that a long residency of Y. pestis of nearly 100 years in the enzootic plague foci has resulted in shedding of virulence genes in the LCR plasmid region in a fairly large proportion of the organisms, possibly due to natural recombination.

SU-F-19A-02: Comparison of Absorbed Dose to Water Standards for HDR Ir-192 Brachytherapy Between the LCR, Brazil and NRC, Canada

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

Salata, C; David, M; Almeida, C de

2014-06-15

Purpose: To compare absorbed dose to water standards for HDR brachytherapy dosimetry developed by the Radiological Science Laboratory of Rio de Janeiro State University (LCR) and the National Research Council, Canada (NRC). Methods: The two institutions have separately developed absorbed dose standards based on the Fricke dosimetry system. There are important differences between the two standards, including: preparation and read-out of the Fricke solution, irradiation geometry of the Fricke holder in relation to the Ir-192 source, and determination of the G-value to be used at Ir-192 energies. All measurements for both standards were made directly at the NRC laboratory (i.e.,more » no transfer instrument was used) using a single Ir-192 source (microSelectron v2). In addition, the NRC group has established a self-consistent method to determine the G-value for Ir-192, based on an interpolation between G-values obtained at Co-60 and 250kVp X-rays, and this measurement was repeated using the LCR Fricke solution to investigate possible systematic uncertainties. Results: G-values for Co-60 and 250 kVp x-rays, obtained using the LCR Fricke system, agreed with the NRC values within 0.5 % and 1 % respectively, indicating that the general assumption of universal G-values is appropriate in this case. The standard uncertainty in the determination of G for Ir-192 is estimated to be 0.6 %. For the comparison of absorbed dose measurements at the reference point for Ir-192 (1 cm depth in water, perpendicular to the seed long-axis), the ratio Dw(NRC)/Dw(LCR) was found to be 1.011 with a combined standard uncertainty of 1.7 %, k=1. Conclusion: The agreement in the absorbed dose to water values for the LCR and NRC systems is very encouraging. Combined with the lower uncertainty in this approach compared to the present air-kerma approach, these results reaffirm the use of Fricke solution as a potential primary standard for HDR Ir-192 brachytherapy.« less

Lateral cephalometric analysis for treatment planning in orthodontics based on MRI compared with radiographs: A feasibility study in children and adolescents

PubMed Central

Lazo Gonzalez, Eduardo; Hilgenfeld, Tim; Kickingereder, Philipp; Bendszus, Martin; Heiland, Sabine; Ozga, Ann-Kathrin; Sommer, Andreas; Lux, Christopher J.; Zingler, Sebastian

2017-01-01

Objective The objective of this prospective study was to evaluate whether magnetic resonance imaging (MRI) is equivalent to lateral cephalometric radiographs (LCR, “gold standard”) in cephalometric analysis. Methods The applied MRI technique was optimized for short scanning time, high resolution, high contrast and geometric accuracy. Prior to orthodontic treatment, 20 patients (mean age ± SD, 13.95 years ± 5.34) received MRI and LCR. MRI datasets were postprocessed into lateral cephalograms. Cephalometric analysis was performed twice by two independent observers for both modalities with an interval of 4 weeks. Eight bilateral and 10 midsagittal landmarks were identified, and 24 widely used measurements (14 angles, 10 distances) were calculated. Statistical analysis was performed by using intraclass correlation coefficient (ICC), Bland-Altman analysis and two one-sided tests (TOST) within the predefined equivalence margin of ± 2°/mm. Results Geometric accuracy of the MRI technique was confirmed by phantom measurements. Mean intraobserver ICC were 0.977/0.975 for MRI and 0.975/0.961 for LCR. Average interobserver ICC were 0.980 for MRI and 0.929 for LCR. Bland-Altman analysis showed high levels of agreement between the two modalities, bias range (mean ± SD) was -0.66 to 0.61 mm (0.06 ± 0.44) for distances and -1.33 to 1.14° (0.06 ± 0.71) for angles. Except for the interincisal angle (p = 0.17) all measurements were statistically equivalent (p < 0.05). Conclusions This study demonstrates feasibility of orthodontic treatment planning without radiation exposure based on MRI. High-resolution isotropic MRI datasets can be transformed into lateral cephalograms allowing reliable measurements as applied in orthodontic routine with high concordance to the corresponding measurements on LCR. PMID:28334054

In Vivo Laser Cartilage Reshaping with Carbon Dioxide Spray Cooling in a Rabbit Ear Model: A Pilot Study

PubMed Central

Kuan, Edward C.; Hamamoto, Ashley A.; Sun, Victor; Nguyen, Tony; Manuel, Cyrus T.; Protsenko, Dmitry E.; Wong, Brian J.F.; Nelson, J. Stuart; Jia, Wangcun

2014-01-01

BACKGROUND/OBJECTIVES Similar to conventional cryogen spray cooling, carbon dioxide (CO2) spray may be used in combination with laser cartilage reshaping (LCR) to produce cartilage shape change while minimizing cutaneous thermal injury. Recent ex vivo evaluation of LCR with CO2 cooling in a rabbit model has identified a promising initial parameter space for in vivo safety and efficacy evaluation. This pilot study aimed to evaluate shape change and cutaneous injury following LCR with CO2 cooling in 5 live rabbits. STUDY DESIGN/MATERIALS AND METHODS The midportion of live rabbit ears were irradiated with a 1.45 μm wavelength diode laser (12 J/cm2) with simultaneous CO2 spray cooling (85 ms duration, 4 alternating heating/cooling cycles per site, 5 to 6 irradiation sites per row for 3 rows per ear). Experimental and control ears (no LCR) were splinted in the flexed position for 30 days following exposure. A total of 5 ears each were allocated to the experimental and control groups. RESULTS Shape change was observed in all irradiated ears (mean 70 ± 3°), which was statistically different from control (mean 37 ± 11 °, p = 0.009). No significant thermal cutaneous injury was observed, with preservation of the full thickness of skin, microvasculature, and adnexal structures. Confocal microscopy and histology demonstrated an intact and viable chondrocyte population surrounding irradiated sites. CONCLUSIONS LCR with CO2 spray cooling can produce clinically significant shape change in the rabbit auricle while minimizing thermal cutaneous and cartilaginous injury and frostbite. This pilot study lends support for the potential use of CO2 spray as an adjunct to existing thermal-based cartilage reshaping modalities. An in vivo systematic evaluation of optimal laser dosimetry and cooling parameters is required. PMID:25557008

Chemiluminescence resonance energy transfer imaging on magnetic particles for single-nucleotide polymorphism detection based on ligation chain reaction.

PubMed

Bi, Sai; Zhang, Zhipeng; Dong, Ying; Wang, Zonghua

2015-03-15

A novel ligation chain reaction (LCR) methodology for single-nucleotide polymorphism (SNP) detection was developed based on luminol-H2O2-horseradish peroxidase (HRP)-mimicking DNAzyme-fluorescein chemiluminescence resonance energy transfer (CRET) imaging on magnetic particles. For LCR, four unique target-complement probes (X and X(⁎), YG and Y(⁎)) for the amplification of K-ras (G12C) were designed by modifying G-quadruplex sequence at 3'-end of YG and fluorescein at 5'-end of Y(⁎). After the LCR, the resulting products of XYG/X(⁎)Y(⁎) with biotin-labeled X(⁎) were captured onto streptavidin-coated magnetic particles (SA-MPs) via specific biotin-SA interaction, which stimulated the CRET reaction from hemin/G-quadruplex-catalyzed luminol-H2O2 CL system to fluorescein. By collecting signals by a cooled low-light CCD, a CRET imaging method was proposed for visual detection and quantitative analysis of SNP. As low as 0.86fM mutant DNA was detected by this assay, and positive mutation detection was achieved with a wild-type to mutant ratio of 10,000:1. This high sensitivity and specificity could be attributed to not only the exponential amplification and excellent discrimination of LCR but also the employment of SA-MPs. SA-MPs ensured the feasibility of the proposed strategy, which also simplified the operations through magnetic separation and separated the reaction and detection procedures to improve sensitivity. The proposed LCR-CRET imaging strategy extends the application of signal amplification techniques to SNP detection, providing a promising platform for effective and high-throughput genetic diagnosis. Copyright © 2014 Elsevier B.V. All rights reserved.

Lateral cephalometric analysis for treatment planning in orthodontics based on MRI compared with radiographs: A feasibility study in children and adolescents.

PubMed

Heil, Alexander; Lazo Gonzalez, Eduardo; Hilgenfeld, Tim; Kickingereder, Philipp; Bendszus, Martin; Heiland, Sabine; Ozga, Ann-Kathrin; Sommer, Andreas; Lux, Christopher J; Zingler, Sebastian

2017-01-01

The objective of this prospective study was to evaluate whether magnetic resonance imaging (MRI) is equivalent to lateral cephalometric radiographs (LCR, "gold standard") in cephalometric analysis. The applied MRI technique was optimized for short scanning time, high resolution, high contrast and geometric accuracy. Prior to orthodontic treatment, 20 patients (mean age ± SD, 13.95 years ± 5.34) received MRI and LCR. MRI datasets were postprocessed into lateral cephalograms. Cephalometric analysis was performed twice by two independent observers for both modalities with an interval of 4 weeks. Eight bilateral and 10 midsagittal landmarks were identified, and 24 widely used measurements (14 angles, 10 distances) were calculated. Statistical analysis was performed by using intraclass correlation coefficient (ICC), Bland-Altman analysis and two one-sided tests (TOST) within the predefined equivalence margin of ± 2°/mm. Geometric accuracy of the MRI technique was confirmed by phantom measurements. Mean intraobserver ICC were 0.977/0.975 for MRI and 0.975/0.961 for LCR. Average interobserver ICC were 0.980 for MRI and 0.929 for LCR. Bland-Altman analysis showed high levels of agreement between the two modalities, bias range (mean ± SD) was -0.66 to 0.61 mm (0.06 ± 0.44) for distances and -1.33 to 1.14° (0.06 ± 0.71) for angles. Except for the interincisal angle (p = 0.17) all measurements were statistically equivalent (p < 0.05). This study demonstrates feasibility of orthodontic treatment planning without radiation exposure based on MRI. High-resolution isotropic MRI datasets can be transformed into lateral cephalograms allowing reliable measurements as applied in orthodontic routine with high concordance to the corresponding measurements on LCR.

A Replication-Defective Human Type 5 Adenovirus-Based Trivalent Vaccine Confers Complete Protection against Plague in Mice and Nonhuman Primates

PubMed Central

Kirtley, Michelle L.; Klages, Curtis; Erova, Tatiana E.; Telepnev, Maxim; Ponnusamy, Duraisamy; Fitts, Eric C.; Baze, Wallace B.; Sivasubramani, Satheesh K.; Lawrence, William S.; Patrikeev, Igor; Peel, Jennifer E.; Andersson, Jourdan A.; Kozlova, Elena V.; Tiner, Bethany L.; Peterson, Johnny W.; McWilliams, David; Patel, Snehal; Rothe, Eric; Motin, Vladimir L.

2016-01-01

Currently, no plague vaccine exists in the United States for human use. The capsular antigen (Caf1 or F1) and two type 3 secretion system (T3SS) components, the low-calcium-response V antigen (LcrV) and the needle protein YscF, represent protective antigens of Yersinia pestis. We used a replication-defective human type 5 adenovirus (Ad5) vector and constructed recombinant monovalent and trivalent vaccines (rAd5-LcrV and rAd5-YFV) that expressed either the codon-optimized lcrV or the fusion gene designated YFV (consisting of ycsF, caf1, and lcrV). Immunization of mice with the trivalent rAd5-YFV vaccine by either the intramuscular (i.m.) or the intranasal (i.n.) route provided protection superior to that with the monovalent rAd5-LcrV vaccine against bubonic and pneumonic plague when animals were challenged with Y. pestis CO92. Preexisting adenoviral immunity did not diminish the protective response, and the protection was always higher when mice were administered one i.n. dose of the trivalent vaccine (priming) followed by a single i.m. booster dose of the purified YFV antigen. Immunization of cynomolgus macaques with the trivalent rAd5-YFV vaccine by the prime-boost strategy provided 100% protection against a stringent aerosol challenge dose of CO92 to animals that had preexisting adenoviral immunity. The vaccinated and challenged macaques had no signs of disease, and the invading pathogen rapidly cleared with no histopathological lesions. This is the first report showing the efficacy of an adenovirus-vectored trivalent vaccine against pneumonic plague in mouse and nonhuman primate (NHP) models. PMID:27170642

A Replication-Defective Human Type 5 Adenovirus-Based Trivalent Vaccine Confers Complete Protection against Plague in Mice and Nonhuman Primates.

PubMed

Sha, Jian; Kirtley, Michelle L; Klages, Curtis; Erova, Tatiana E; Telepnev, Maxim; Ponnusamy, Duraisamy; Fitts, Eric C; Baze, Wallace B; Sivasubramani, Satheesh K; Lawrence, William S; Patrikeev, Igor; Peel, Jennifer E; Andersson, Jourdan A; Kozlova, Elena V; Tiner, Bethany L; Peterson, Johnny W; McWilliams, David; Patel, Snehal; Rothe, Eric; Motin, Vladimir L; Chopra, Ashok K

2016-07-01

Currently, no plague vaccine exists in the United States for human use. The capsular antigen (Caf1 or F1) and two type 3 secretion system (T3SS) components, the low-calcium-response V antigen (LcrV) and the needle protein YscF, represent protective antigens of Yersinia pestis We used a replication-defective human type 5 adenovirus (Ad5) vector and constructed recombinant monovalent and trivalent vaccines (rAd5-LcrV and rAd5-YFV) that expressed either the codon-optimized lcrV or the fusion gene designated YFV (consisting of ycsF, caf1, and lcrV). Immunization of mice with the trivalent rAd5-YFV vaccine by either the intramuscular (i.m.) or the intranasal (i.n.) route provided protection superior to that with the monovalent rAd5-LcrV vaccine against bubonic and pneumonic plague when animals were challenged with Y. pestis CO92. Preexisting adenoviral immunity did not diminish the protective response, and the protection was always higher when mice were administered one i.n. dose of the trivalent vaccine (priming) followed by a single i.m. booster dose of the purified YFV antigen. Immunization of cynomolgus macaques with the trivalent rAd5-YFV vaccine by the prime-boost strategy provided 100% protection against a stringent aerosol challenge dose of CO92 to animals that had preexisting adenoviral immunity. The vaccinated and challenged macaques had no signs of disease, and the invading pathogen rapidly cleared with no histopathological lesions. This is the first report showing the efficacy of an adenovirus-vectored trivalent vaccine against pneumonic plague in mouse and nonhuman primate (NHP) models. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Laparoscopic colon resection trends in utilization and rate of conversion to open procedure: a national database review of academic medical centers.

PubMed

Simorov, Anton; Shaligram, Abhijit; Shostrom, Valerie; Boilesen, Eugene; Thompson, Jon; Oleynikov, Dmitry

2012-09-01

This study aims to examine trends of utilization and rates of conversion to open procedure for patients undergoing laparoscopic colon resections (LCR). This study is a national database review of academic medical centers and a retrospective analysis utilizing the University HealthSystem Consortium administrative database-an alliance of more than 300 academic and affiliate hospitals. A total of 85,712 patients underwent colon resections between October 2008 and December 2011. LCR was attempted in 36,228 patients (42.2%), with 5751 patients (15.8%) requiring conversion to an open procedure. There was a trend toward increasing utilization of LCR from 37.5% in 2008 to 44.1% in 2011. Attempted laparoscopic transverse colectomy had the highest rate of conversion (20.8%), followed by left (20.7%), right (15.6%), and sigmoid (14.3%) colon resections. The rate of utilization was highest in the Mid-Atlantic region (50.5%) and in medium- to large-sized hospitals (47.0%-49.0%).Multivariate logistic regression has shown that increasing age [odds ratio (OR) = 4.8, 95% confidence interval (CI) = 3.6-6.4], male sex (OR = 1.2, 95% CI = 1.1-1.3), open as compared with laparoscopic approach (OR = 2.6, 95%, CI = 2.3-3.1), and greater severity of illness category (OR = 27.1, 95% CI = 23.0-31.9) were all associated with increased mortality and morbidity and prolonged length of hospital stay. There is a trend of increasing utilization of LCR, with acceptable conversion rates, across hospitals in the United States over the recent years. When feasible, attempted LCR had better outcomes than open colectomy in the immediate perioperative period.

Design of a 9-loop quasi-exponential waveform generator

NASA Astrophysics Data System (ADS)

Banerjee, Partha; Shukla, Rohit; Shyam, Anurag

2015-12-01

We know in an under-damped L-C-R series circuit, current follows a damped sinusoidal waveform. But if a number of sinusoidal waveforms of decreasing time period, generated in an L-C-R circuit, be combined in first quarter cycle of time period, then a quasi-exponential nature of output current waveform can be achieved. In an L-C-R series circuit, quasi-exponential current waveform shows a rising current derivative and thereby finds many applications in pulsed power. Here, we have described design and experiment details of a 9-loop quasi-exponential waveform generator. In that, design details of magnetic switches have also been described. In the experiment, output current of 26 kA has been achieved. It has been shown that how well the experimentally obtained output current profile matches with the numerically computed output.

Design of a 9-loop quasi-exponential waveform generator.

PubMed

Banerjee, Partha; Shukla, Rohit; Shyam, Anurag

2015-12-01

We know in an under-damped L-C-R series circuit, current follows a damped sinusoidal waveform. But if a number of sinusoidal waveforms of decreasing time period, generated in an L-C-R circuit, be combined in first quarter cycle of time period, then a quasi-exponential nature of output current waveform can be achieved. In an L-C-R series circuit, quasi-exponential current waveform shows a rising current derivative and thereby finds many applications in pulsed power. Here, we have described design and experiment details of a 9-loop quasi-exponential waveform generator. In that, design details of magnetic switches have also been described. In the experiment, output current of 26 kA has been achieved. It has been shown that how well the experimentally obtained output current profile matches with the numerically computed output.

The 'real-world' impact of improved diagnostic techniques for Chlamydia trachomatis infection in Glasgow.

PubMed

Scoular, A; McCartney, R; Kinn, S; Carr, S; Walker, A

2001-09-01

In April 1997, the main chlamydia laboratory in Glasgow introduced ligase chain reaction (LCR) as its standard diagnostic test. The diagnostic effectiveness and health economic impact of introduction of LCR testing was assessed. Between April 1996 to March 2000, results of all chlamydia detection tests on genital specimens sent from general practitioners and the two main sexual healthcare providers (Genitourinary Medicine and Family Planning services) were reviewed. A preliminary economic assessment, inclusive of staff, reagents, consumables and laboratory overheads was conducted. Overall, testing activity increased four and a half times between 1996-97 and 1999-2000; the proportionate rise was greatest in general practice. Although chlamydia testing in both genders increased over the review period, testing activity rose disproportionately in women (59%, compared with a 31% increase in men). The overall Chlamydia trachomatis detection rate rose from 4.8% in 1996-97 to 7.8% in 1999-2000. Following introduction of LCR testing, an estimated additional 331 men and 844 women were diagnosed during the study period. The cost per additional diagnosis made was estimated at 162 Pounds for men and 263 Pounds for women. Substantial health gains are likely to be achieved, at both an individual and public health level, as a result of introduction of LCR testing for genital chlamydial infection.

Efficacy and toxicity profiles of two chemoradiotherapies for stage II laryngeal cancer - a comparison between late course accelerated hyperfractionation (LCAHF) and conventional fractionation (CF).

PubMed

Okazaki, Eiichiro; Matsushita, Naoki; Tashiro, Mari; Shimatani, Yasuhiko; Ishii, Kentaro; Hosono, Masako; Oishi, Masahiro; Teranishi, Yuichi; Iguchi, Hiroyoshi; Miki, Yukio

2017-08-01

To evaluate the treatment results of late course accelerated hyperfractionation (LCAHF) compared with conventional fractionation (CF) for stage II laryngeal cancer. Fifty-nine consecutive patients treated for stage II laryngeal cancer were retrospectively reviewed. Thirty-two patients underwent LCAHF, twice-daily fractions during the latter half with a total dose of 69 Gy. Twenty-seven patients received CF of 70 Gy. The local control rates (LCRs), overall survival (OS), and disease-specific survival (DSS) at 5 years were 80.6%, 74.0%, and 90.4%, respectively, after LCAHF and 64.7%, 68.2%, and 90.5%, respectively, after CF. There were no significant differences in LCR, OS, and DSS (p = .11, 0.68, and 0.69, respectively). In a small number of patients with supraglottic cancer, LCAHF was associated with a significantly higher LCR at 5 years compared with CF (100% vs. 41.7%; p = .02). This is the first report that compared the results of LCAHF and CF for stage II laryngeal cancer. We could not find significant differences in LCR, DSS, and OS rates between LCAHF and CF groups. Although in a small number of patients with supraglottic cancer, LCAHF may improve the LCR compared with CF.

Differences in Transcriptional Activity of Human Papillomavirus Type 6 Molecular Variants in Recurrent Respiratory Papillomatosis

PubMed Central

Measso do Bonfim, Caroline; Simão Sobrinho, João; Lacerda Nogueira, Rodrigo; Salgado Kupper, Daniel; Cardoso Pereira Valera, Fabiana; Lacerda Nogueira, Maurício; Villa, Luisa Lina; Rahal, Paula; Sichero, Laura

2015-01-01

A significant proportion of recurrent respiratory papillomatosis (RRP) is caused by human papillomavirus type 6 (HPV-6). The long control region (LCR) contains cis-elements for regulation of transcription. Our aim was to characterize LCR HPV-6 variants in RRP cases, compare promoter activity of these isolates and search for cellular transcription factors (TFs) that could explain the differences observed. The complete LCR from 13 RRP was analyzed. Transcriptional activity of 5 variants was compared using luciferase assays. Differences in putative TFs binding sites among variants were revealed using the TRANSFAC database. Chromatin immunoprecipation (CHIP) and luciferase assays were used to evaluate TF binding and impact upon transcription, respectively. Juvenile-onset RRP cases harbored exclusively HPV-6vc related variants, whereas among adult-onset cases HPV-6a variants were more prevalent. The HPV-6vc reference was more transcriptionally active than the HPV-6a reference. Active FOXA1, ELF1 and GATA1 binding sites overlap variable nucleotide positions among isolates and influenced LCR activity. Furthermore, our results support a crucial role for ELF1 on transcriptional downregulation. We identified TFs implicated in the regulation of HPV-6 early gene expression. Many of these factors are mutated in cancer or are putative cancer biomarkers, and must be further studied. PMID:26151558

High Fractional Occupancy of a Tandem Maf Recognition Element and Its Role in Long-Range β-Globin Gene Regulation

PubMed Central

Stees, Jared R.; Hossain, Mir A.; Sunose, Tomoki; Kudo, Yasushi; Pardo, Carolina E.; Nabilsi, Nancy H.; Darst, Russell P.; Poudyal, Rosha; Igarashi, Kazuhiko; Kladde, Michael P.

2015-01-01

Enhancers and promoters assemble protein complexes that ultimately regulate the recruitment and activity of RNA polymerases. Previous work has shown that at least some enhancers form stable protein complexes, leading to the formation of enhanceosomes. We analyzed protein-DNA interactions in the murine β-globin gene locus using the methyltransferase accessibility protocol for individual templates (MAPit). The data show that a tandem Maf recognition element (MARE) in locus control region (LCR) hypersensitive site 2 (HS2) reveals a remarkably high degree of occupancy during differentiation of mouse erythroleukemia cells. Most of the other transcription factor binding sites in LCR HS2 or in the adult β-globin gene promoter regions exhibit low fractional occupancy, suggesting highly dynamic protein-DNA interactions. Targeting of an artificial zinc finger DNA-binding domain (ZF-DBD) to the HS2 tandem MARE caused a reduction in the association of MARE-binding proteins and transcription complexes at LCR HS2 and the adult βmajor-globin gene promoter but did not affect expression of the βminor-globin gene. The data demonstrate that a stable MARE-associated footprint in LCR HS2 is important for the recruitment of transcription complexes to the adult βmajor-globin gene promoter during erythroid cell differentiation. PMID:26503787

Yersinia pestis caf1 variants and the limits of plague vaccine protection.

PubMed

Quenee, Lauriane E; Cornelius, Claire A; Ciletti, Nancy A; Elli, Derek; Schneewind, Olaf

2008-05-01

Yersinia pestis, the highly virulent agent of plague, is a biological weapon. Strategies that prevent plague have been sought for centuries, and immunization with live, attenuated (nonpigmented) strains or subunit vaccines with F1 (Caf1) antigen is considered effective. We show here that immunization with live, attenuated strains generates plague-protective immunity and humoral immune responses against F1 pilus antigen and LcrV. Y. pestis variants lacking caf1 (F1 pili) are not only fully virulent in animal models of bubonic and pneumonic plague but also break through immune responses generated with live, attenuated strains or F1 subunit vaccines. In contrast, immunization with purified LcrV, a protein at the tip of type III needles, generates protective immunity against the wild-type and the fully virulent caf1 mutant strain, in agreement with the notion that LcrV can elicit vaccine protection against both types of virulent plague strains.

Proton, muon and ¹³C hyperfine coupling constants of C₆₀X and C₇₀X (X = H, Mu).

PubMed

Brodovitch, Jean-Claude; Addison-Jones, Brenda; Ghandi, Khashayar; McKenzie, Iain; Percival, Paul W

2015-01-21

The reaction of H atoms with fullerene C70 has been investigated by identifying the radical products formed by addition of the atom muonium (Mu) to the fullerene in solution. Four of the five possible radical isomers of C70Mu were detected by avoided level-crossing resonance (μLCR) spectroscopy, using a dilute solution of enriched (13)C70 in decalin. DFT calculations were used to predict muon and (13)C isotropic hyperfine constants as an aid to assigning the observed μLCR signals. Computational methods were benchmarked against previously published experimental data for (13)C60Mu in solution. Analysis of the μLCR spectrum resulted in the first experimental determination of (13)C hyperfine constants in either C70Mu or C70H. The large number of values confirms predictions that the four radical isomers have extended distributions of unpaired electron spin.

The C-Terminal Domain of Nrf1 Negatively Regulates the Full-Length CNC-bZIP Factor and Its Shorter Isoform LCR-F1/Nrf1β; Both Are Also Inhibited by the Small Dominant-Negative Nrf1γ/δ Isoforms that Down-Regulate ARE-Battery Gene Expression

PubMed Central

Zhang, Yiguo; Qiu, Lu; Li, Shaojun; Xiang, Yuancai; Chen, Jiayu; Ren, Yonggang

2014-01-01

The C-terminal domain (CTD, aa 686–741) of nuclear factor-erythroid 2 p45-related factor 1 (Nrf1) shares 53% amino acid sequence identity with the equivalent Neh3 domain of Nrf2, a homologous transcription factor. The Neh3 positively regulates Nrf2, but whether the Neh3-like (Neh3L) CTD of Nrf1 has a similar role in regulating Nrf1-target gene expression is unknown. Herein, we report that CTD negatively regulates the full-length Nrf1 (i.e. 120-kDa glycoprotein and 95-kDa deglycoprotein) and its shorter isoform LCR-F1/Nrf1β (55-kDa). Attachment of its CTD-adjoining 112-aa to the C-terminus of Nrf2 yields the chimaeric Nrf2-C112Nrf1 factor with a markedly decreased activity. Live-cell imaging of GFP-CTD reveals that the extra-nuclear portion of the fusion protein is allowed to associate with the endoplasmic reticulum (ER) membrane through the amphipathic Neh3L region of Nrf1 and its basic c-tail. Thus removal of either the entire CTD or the essential Neh3L portion within CTD from Nrf1, LCR-F1/Nrf1β and Nrf2-C112Nrf1, results in an increase in their transcriptional ability to regulate antioxidant response element (ARE)-driven reporter genes. Further examinations unravel that two smaller isoforms, 36-kDa Nrf1γ and 25-kDa Nrf1δ, act as dominant-negative inhibitors to compete against Nrf1, LCR-F1/Nrf1β and Nrf2. Relative to Nrf1, LCR-F1/Nrf1β is a weak activator, that is positively regulated by its Asn/Ser/Thr-rich (NST) domain and acidic domain 2 (AD2). Like AD1 of Nrf1, both AD2 and NST domain of LCR-F1/Nrf1β fused within two different chimaeric contexts to yield Gal4D:Nrf1β607 and Nrf1β:C270Nrf2, positively regulate their transactivation activity of cognate Gal4- and Nrf2-target reporter genes. More importantly, differential expression of endogenous ARE-battery genes is attributable to up-regulation by Nrf1 and LCR-F1/Nrf1β and down-regulation by Nrf1γ and Nrf1δ. PMID:25290918

Immunogenicity and protective immunity against bubonic plague and pneumonic plague by immunization of mice with the recombinant V10 antigen, a variant of LcrV.

PubMed

DeBord, Kristin L; Anderson, Deborah M; Marketon, Melanie M; Overheim, Katie A; DePaolo, R William; Ciletti, Nancy A; Jabri, Bana; Schneewind, Olaf

2006-08-01

In contrast to Yersinia pestis LcrV, the recombinant V10 (rV10) variant (lacking residues 271 to 300) does not suppress the release of proinflammatory cytokines by immune cells. Immunization with rV10 generates robust antibody responses that protect mice against bubonic plague and pneumonic plague, suggesting that rV10 may serve as an improved plague vaccine.

A Yersinia pestis YscN ATPase mutant functions as a live attenuated vaccine against bubonic plague in mice.

PubMed

Bozue, Joel; Cote, Christopher K; Webster, Wendy; Bassett, Anthony; Tobery, Steven; Little, Stephen; Swietnicki, Wieslaw

2012-07-01

Yersinia pestis is the causative agent responsible for bubonic and pneumonic plague. The bacterium uses the pLcr plasmid-encoded type III secretion system to deliver virulence factors into host cells. Delivery requires ATP hydrolysis by the YscN ATPase encoded by the yscN gene also on pLcr. A yscN mutant was constructed in the fully virulent CO92 strain containing a nonpolar, in-frame internal deletion within the gene. We demonstrate that CO92 with a yscN mutation was not able to secrete the LcrV protein (V-Antigen) and attenuated in a subcutaneous model of plague demonstrating that the YscN ATPase was essential for virulence. However, if the yscN mutant was complemented with a functional yscN gene in trans, virulence was restored. To evaluate the mutant as a live vaccine, Swiss-Webster mice were vaccinated twice with the ΔyscN mutant at varying doses and were protected against bubonic plague in a dose-dependent manner. Antibodies to F1 capsule but not to LcrV were detected in sera from the vaccinated mice. These preliminary results suggest a proof-of-concept for an attenuated, genetically engineered, live vaccine effective against bubonic plague. Published 2012. This article is a US Government work and is in the public domain in the USA.

Plague in Guinea Pigs and Its Prevention by Subunit Vaccines

PubMed Central

Quenee, Lauriane E.; Ciletti, Nancy; Berube, Bryan; Krausz, Thomas; Elli, Derek; Hermanas, Timothy; Schneewind, Olaf

2011-01-01

Human pneumonic plague is a devastating and transmissible disease for which a Food and Drug Administration–approved vaccine is not available. Suitable animal models may be adopted as a surrogate for human plague to fulfill regulatory requirements for vaccine efficacy testing. To develop an alternative to pneumonic plague in nonhuman primates, we explored guinea pigs as a model system. On intranasal instillation of a fully virulent strain, Yersinia pestis CO92, guinea pigs developed lethal lung infections with hemorrhagic necrosis, massive bacterial replication in the respiratory system, and blood-borne dissemination to other organ systems. Expression of the Y. pestis F1 capsule was not required for the development of pulmonary infection; however, the capsule seemed to be important for the establishment of bubonic plague. The mean lethal dose (MLD) for pneumonic plague in guinea pigs was estimated to be 1000 colony-forming units. Immunization of guinea pigs with the recombinant forms of LcrV, a protein that resides at the tip of Yersinia type III secretion needles, or F1 capsule generated robust humoral immune responses. Whereas LcrV immunization resulted in partial protection against pneumonic plague challenge with 250 MLD Y. pestis CO92, immunization with recombinant F1 did not. rV10, a vaccine variant lacking LcrV residues 271-300, elicited protection against pneumonic plague, which seemed to be based on conformational antibodies directed against LcrV. PMID:21406168

Real-time monitoring of stress erythropoiesis in vivo using Gata1 and beta-globin LCR luciferase transgenic mice.

PubMed

Suzuki, Mikiko; Ohneda, Kinuko; Hosoya-Ohmura, Sakie; Tsukamoto, Saho; Ohneda, Osamu; Philipsen, Sjaak; Yamamoto, Masayuki

2006-07-15

Erythroid progenitors have the potential to proliferate rapidly in response to environmental stimuli. This process is referred to as stress erythropoiesis, with erythropoietin (EPO) playing central roles in its promotion. In this study, we wanted to elucidate the molecular mechanisms governing the regulation of stress erythropoiesis and the maintenance of red-cell homeostasis. This was achieved by our development of a noninvasive real-time monitoring system for erythropoiesis using transgenic mouse lines expressing luciferase under the control of the mouse Gata1 hematopoietic regulatory domain (G1-HRD-luc) or human beta-globin locus control region (Hbb-LCR-luc). Optical bioluminescence images revealed that the luciferase was specifically expressed in spleen and bone marrow and was induced rapidly in response to anemia and hypoxia stimuli. The G1-HRD-luc activity tracked the emergence and disappearance of proerythroblast-stage progenitors, whereas the Hbb-LCR-luc activity tracked erythroblasts and later stage erythroid cells. Increased plasma EPO concentration preceded an increase in G1-HRD-luc, supporting our contention that EPO acts as the key upstream signal in stress erythropoiesis. Hence, we conclude that G1-HRD-luc and Hbb-LCR-luc reporters are differentially activated during stress erythropoiesis and that the transgenic mouse lines used serve as an important means for understanding the homeostatic regulation of erythropoiesis.

Numerical investigations of internal stresses on carbon steel based on ultrasonic LCR waves

NASA Astrophysics Data System (ADS)

Ramasamy, R.; Ibrahim, Z.; Chai, H. K.

2017-10-01

Internal stresses or residual stresses in the structural elements are very crucial in carrying out in-service evaluations and fitness-for-purpose assessments. The generation of these internal stresses can occur as result of the fabrication of the steel members, installation sequence or other ad-hoc events such as accidents or impact. The accurate prediction of the internal stresses will contribute towards estimating the integrity state of the structural elements, with respect to their material allowable stresses. This paper investigates the explicit FE based numerical modelling of the ultrasonic based non-destructive technique, utilising the measurable longitudinal critical refracted wave (LCR) and relating these to the internal stresses within the structural elements by the evaluation of the material dependent acoustoelastic factors. The subsurface travel path of the LCR wave inside the structural elements makes it a sub-surface stress measurement technique and the linearised relationship with corresponding internal stresses can be systematically applied repeatedly. The numerical results are compared against laboratory tests data to correlate the findings and to establish modelling feasibility for future proof-of-concepts. It can be concluded from this numerical investigation, that the subsurface ultrasonic LCR wave has great potential to be implemented for in-situ structural residual stress measurements, as compared to other available surface measurements such as strain gauges or x-ray diffraction.

Relation between acute and long-term cognitive decline after surgery: Influence of metabolic syndrome.

PubMed

Gambús, P L; Trocóniz, I F; Feng, X; Gimenez-Milá, M; Mellado, R; Degos, V; Vacas, S; Maze, M

2015-11-01

The relationship between persistent postoperative cognitive decline and the more common acute variety remains unknown; using data acquired in preclinical studies of postoperative cognitive decline we attempted to characterize this relationship. Low capacity runner (LCR) rats, which have all the features of the metabolic syndrome, were compared postoperatively with high capacity runner (HCR) rats for memory, assessed by trace fear conditioning (TFC) on the 7th postoperative day, and learning and memory (probe trial [PT]) assessed by the Morris water-maze (MWM) at 3 months postoperatively. Rate of learning (AL) data from the MWM test, were estimated by non-linear mixed effects modeling. The individual rat's TFC result at postoperative day (POD) 7 was correlated with its AL and PT from the MWM data sets at postoperative day POD 90. A single exponential decay model best described AL in the MWM with LCR and surgery (LCR-SURG) being the only significant covariates; first order AL rate constant was 0.07 s(-1) in LCR-SURG and 0.16s(-1) in the remaining groups (p<0.05). TFC was significantly correlated with both AL (R=0.74; p<0.0001) and PT (R=0.49; p<0.01). Severity of memory decline at 1 week after surgery presaged long-lasting deteriorations in learning and memory. Copyright © 2015 Elsevier Inc. All rights reserved.

Computer algorithms for automated detection and analysis of local Ca2+ releases in spontaneously beating cardiac pacemaker cells

PubMed Central

Kim, Mary S.; Tsutsui, Kenta; Stern, Michael D.; Lakatta, Edward G.; Maltsev, Victor A.

2017-01-01

Local Ca2+ Releases (LCRs) are crucial events involved in cardiac pacemaker cell function. However, specific algorithms for automatic LCR detection and analysis have not been developed in live, spontaneously beating pacemaker cells. In the present study we measured LCRs using a high-speed 2D-camera in spontaneously contracting sinoatrial (SA) node cells isolated from rabbit and guinea pig and developed a new algorithm capable of detecting and analyzing the LCRs spatially in two-dimensions, and in time. Our algorithm tracks points along the midline of the contracting cell. It uses these points as a coordinate system for affine transform, producing a transformed image series where the cell does not contract. Action potential-induced Ca2+ transients and LCRs were thereafter isolated from recording noise by applying a series of spatial filters. The LCR birth and death events were detected by a differential (frame-to-frame) sensitivity algorithm applied to each pixel (cell location). An LCR was detected when its signal changes sufficiently quickly within a sufficiently large area. The LCR is considered to have died when its amplitude decays substantially, or when it merges into the rising whole cell Ca2+ transient. Ultimately, our algorithm provides major LCR parameters such as period, signal mass, duration, and propagation path area. As the LCRs propagate within live cells, the algorithm identifies splitting and merging behaviors, indicating the importance of locally propagating Ca2+-induced-Ca2+-release for the fate of LCRs and for generating a powerful ensemble Ca2+ signal. Thus, our new computer algorithms eliminate motion artifacts and detect 2D local spatiotemporal events from recording noise and global signals. While the algorithms were developed to detect LCRs in sinoatrial nodal cells, they have the potential to be used in other applications in biophysics and cell physiology, for example, to detect Ca2+ wavelets (abortive waves), sparks and embers in muscle cells and Ca2+ puffs and syntillas in neurons. PMID:28683095

Stabilization of diastolic calcium signal via calcium pump regulation of complex local calcium releases and transient decay in a computational model of cardiac pacemaker cell with individual release channels

PubMed Central

Maltsev, Alexander V.; Maltsev, Victor A.; Stern, Michael D.

2017-01-01

Intracellular Local Ca releases (LCRs) from sarcoplasmic reticulum (SR) regulate cardiac pacemaker cell function by activation of electrogenic Na/Ca exchanger (NCX) during diastole. Prior studies demonstrated the existence of powerful compensatory mechanisms of LCR regulation via a complex local cross-talk of Ca pump, release and NCX. One major obstacle to study these mechanisms is that LCR exhibit complex Ca release propagation patterns (including merges and separations) that have not been characterized. Here we developed new terminology, classification, and computer algorithms for automatic detection of numerically simulated LCRs and examined LCR regulation by SR Ca pumping rate (Pup) that provides a major contribution to fight-or-flight response. In our simulations the faster SR Ca pumping accelerates action potential-induced Ca transient decay and quickly clears Ca under the cell membrane in diastole, preventing premature releases. Then the SR generates an earlier, more synchronized, and stronger diastolic LCR signal activating an earlier and larger inward NCX current. LCRs at higher Pup exhibit larger amplitudes and faster propagation with more collisions to each other. The LCRs overlap with Ca transient decay, causing an elevation of the average diastolic [Ca] nadir to ~200 nM (at Pup = 24 mM/s). Background Ca (in locations lacking LCRs) quickly decays to resting Ca levels (<100 nM) at high Pup, but remained elevated during slower decay at low Pup. Release propagation is facilitated at higher Pup by a larger LCR amplitude, whereas at low Pup by higher background Ca. While at low Pup LCRs show smaller amplitudes, their larger durations and sizes combined with longer transient decay stabilize integrals of diastolic Ca and NCX current signals. Thus, the local interplay of SR Ca pump and release channels regulates LCRs and Ca transient decay to insure fail-safe pacemaker cell operation within a wide range of rates. PMID:28792496

A palindrome-mediated mechanism distinguishes translocations involving LCR-B of chromosome 22q11.2.

PubMed

Gotter, Anthony L; Shaikh, Tamim H; Budarf, Marcia L; Rhodes, C Harker; Emanuel, Beverly S

2004-01-01

Two known recurrent constitutional translocations, t(11;22) and t(17;22), as well as a non-recurrent t(4;22), display derivative chromosomes that have joined to a common site within the low copy repeat B (LCR-B) region of 22q11.2. This breakpoint is located between two AT-rich inverted repeats that form a nearly perfect palindrome. Breakpoints within the 11q23, 17q11 and 4q35 partner chromosomes also fall near the center of palindromic sequences. In the present work the breakpoints of a fourth translocation involving LCR-B, a balanced ependymoma-associated t(1;22), were characterized not only to localize this junction relative to known genes, but also to further understand the mechanism underlying these rearrangements. FISH mapping was used to localize the 22q11.2 breakpoint to LCR-B and the 1p21 breakpoint to single BAC clones. STS mapping narrowed the 1p21.2 breakpoint to a 1990 bp AT-rich region, and junction fragments were amplified by nested PCR. Junction fragment-derived sequence indicates that the 1p21.2 breakpoint splits a 278 nt palindrome capable of forming stem-loop secondary structure. In contrast, the 1p21.2 reference genomic sequence from clones in the database does not exhibit this configuration, suggesting a predisposition for regional genomic instability perhaps etiologic for this rearrangement. Given its similarity to known chromosomal fragile site (FRA) sequences, this polymorphic 1p21.2 sequence may represent one of the FRA1 loci. Comparative analysis of the secondary structure of sequences surrounding translocation breakpoints that involve LCR-B with those not involving this region indicate a unique ability of the former to form stem-loop structures. The relative likelihood of forming these configurations appears to be related to the rate of translocation occurrence. Further analysis suggests that constitutional translocations in general occur between sequences of similar melting temperature and propensity for secondary structure.

Voluntary Running Attenuates Metabolic Dysfunction in Ovariectomized Low-Fit Rats

PubMed Central

Park, Young-Min; Padilla, Jaume; Kanaley, Jill A.; Zidon, Terese; Welly, Rebecca J.; Britton, Steven L.; Koch, Lauren G.; Thyfault, John P.; Booth, Frank W.; Vieira-Potter, Victoria J.

2016-01-01

INTRODUCTION Ovariectomy and high fat diet (HFD) worsen obesity and metabolic dysfunction associated with low aerobic fitness. Exercise training mitigates metabolic abnormalities induced by low aerobic fitness, but whether the protective effect is maintained following ovariectomy and HFD is unknown. PURPOSE This study determined whether, following ovariectomy and HFD, exercise training improves metabolic function in rats bred for low intrinsic aerobic capacity. METHODS Female rats selectively bred for low (LCR) and high (HCR) intrinsic aerobic capacity (n=30) were ovariectomized, fed HFD, and randomized to either a sedentary (SED) or voluntary wheel running (EX) group. Resting energy expenditure, glucose tolerance, and spontaneous physical activity were determined midway through the experiment, while body weight, wheel running volume, and food intake were assessed throughout the study. Body composition, circulating metabolic markers, and skeletal muscle gene and protein expression was measured at sacrifice. RESULTS EX reduced body weight and adiposity in LCR rats (−10% and −50%, respectively; P<0.05) and, unexpectedly, increased these variables in HCR rats (+7% and +37%, respectively; P<0.05) compared to their respective SED controls, likely due to dietary overcompensation. Wheel running volume was ~5-fold greater in HCR than LCR rats, yet EX enhanced insulin sensitivity equally in LCR and HCR rats (P<0.05). This EX-mediated improvement in metabolic function was associated with gene up-regulation of skeletal muscle IL-6&-10. EX also increased resting energy expenditure, skeletal muscle mitochondrial content (oxidative phosphorylation complexes and citrate synthase activity), and AMPK activation similarly in both lines (all P <0.05). CONCLUSION Despite a 5-fold difference in running volume between rat lines, EX similarly improved systemic insulin sensitivity, resting energy expenditure, and skeletal muscle mitochondrial content and AMPK activation in ovariectomized LCR and HCR rats fed HFD compared to their respective SED controls. PMID:27669449

A palindrome-mediated mechanism distinguishes translocations involving LCR-B of chromosome 22q11.2

PubMed Central

Gotter, Anthony L.; Shaikh, Tamim H.; Budarf, Marcia L.; Rhodes, C. Harker; Emanuel, Beverly S.

2010-01-01

Two known recurrent constitutional translocations, t(11;22) and t(17;22), as well as a non-recurrent t(4;22), display derivative chromosomes that have joined to a common site within the low copy repeat B (LCR-B) region of 22q11.2. This breakpoint is located between two AT-rich inverted repeats that form a nearly perfect palindrome. Breakpoints within the 11q23, 17q11 and 4q35 partner chromosomes also fall near the center of palindromic sequences. In the present work the breakpoints of a fourth translocation involving LCR-B, a balanced ependymoma-associated t(1;22), were characterized not only to localize this junction relative to known genes, but also to further understand the mechanism underlying these rearrangements. FISH mapping was used to localize the 22q11.2 breakpoint to LCR-B and the 1p21 breakpoint to single BAC clones. STS mapping narrowed the 1p21.2 breakpoint to a 1990 bp AT-rich region, and junction fragments were amplified by nested PCR. Junction fragment-derived sequence indicates that the 1p21.2 breakpoint splits a 278 nt palindrome capable of forming stem–loop secondary structure. In contrast, the 1p21.2 reference genomic sequence from clones in the database does not exhibit this configuration, suggesting a predisposition for regional genomic instability perhaps etiologic for this rearrangement. Given its similarity to known chromosomal fragile site (FRA) sequences, this polymorphic 1p21.2 sequence may represent one of the FRA1 loci. Comparative analysis of the secondary structure of sequences surrounding translocation breakpoints that involve LCR-B with those not involving this region indicate a unique ability of the former to form stem–loop structures. The relative likelihood of forming these configurations appears to be related to the rate of translocation occurrence. Further analysis suggests that constitutional translocations in general occur between sequences of similar melting temperature and propensity for secondary structure. PMID:14613967

Rat Strains Bred for Low and High Aerobic Running Capacity do not Differ in Their Survival Time to Hemorrhage

DTIC Science & Technology

2009-12-01

23. Ensunsa JL, Symons JD, Lanoue L, Schrader HR, Keen CL. Reducing arginase activity via dietary manganese deficiency enhances endothelium- dependent ...maximal oxygen consumption (Vo2max) was 12% greater during normoxia and 20% greater during hypoxia in the HCR vs LCR (12). This enhanced Vo2max...was ∼ 40% greater in HCR vs LCR due to both increased oxygen delivery-- resulting from an enhanced stroke volume-- and oxygen tissue transfer (13

Robust synchronization of spin-torque oscillators with an LCR load.

PubMed

Pikovsky, Arkady

2013-09-01

We study dynamics of a serial array of spin-torque oscillators with a parallel inductor-capacitor-resistor (LCR) load. In a large range of parameters the fully synchronous regime, where all the oscillators have the same state and the output field is maximal, is shown to be stable. However, not always such a robust complete synchronization develops from a random initial state; in many cases nontrivial clustering is observed, with a partial synchronization resulting in a quasiperiodic or chaotic mean-field dynamics.

The first DNA 1-like alpha satellites in association with New World begomoviruses in natural infections.

PubMed

Paprotka, T; Metzler, V; Jeske, H

2010-09-01

From Brazilian weeds with typical symptoms of a geminivirus infection, the DNAs of two new virus species, two new strains with two variants of already known bipartite begomoviruses were sequenced. Moreover, the first two DNA 1-like satellites (alpha satellites) occurring naturally in the New World were identified. They are related to nanoviral DNA components and show a typical genome organization with one open reading frame coding potentially for a replication-associated protein (Rep), a conserved hairpin structure, and an A-rich region. After coinoculation with their helper begomoviruses (Euphorbia mosaic virus, EuMV or Cleome leaf crumple virus, ClLCrV) the satellite DNAs were transmitted to experimental and natural host plants. Three of the begomovirus isolates (EuMV and ClLCrV) infected Arabidopsis thaliana plants, induced mild symptoms, and one of these (ClLCrV) transreplicated the satellite efficiently. As a result, several novel tools for molecular analyses of this important model plant are provided. 2010 Elsevier Inc. All rights reserved.

Plague in Guinea pigs and its prevention by subunit vaccines.

PubMed

Quenee, Lauriane E; Ciletti, Nancy; Berube, Bryan; Krausz, Thomas; Elli, Derek; Hermanas, Timothy; Schneewind, Olaf

2011-04-01

Human pneumonic plague is a devastating and transmissible disease for which a Food and Drug Administration-approved vaccine is not available. Suitable animal models may be adopted as a surrogate for human plague to fulfill regulatory requirements for vaccine efficacy testing. To develop an alternative to pneumonic plague in nonhuman primates, we explored guinea pigs as a model system. On intranasal instillation of a fully virulent strain, Yersinia pestis CO92, guinea pigs developed lethal lung infections with hemorrhagic necrosis, massive bacterial replication in the respiratory system, and blood-borne dissemination to other organ systems. Expression of the Y. pestis F1 capsule was not required for the development of pulmonary infection; however, the capsule seemed to be important for the establishment of bubonic plague. The mean lethal dose (MLD) for pneumonic plague in guinea pigs was estimated to be 1000 colony-forming units. Immunization of guinea pigs with the recombinant forms of LcrV, a protein that resides at the tip of Yersinia type III secretion needles, or F1 capsule generated robust humoral immune responses. Whereas LcrV immunization resulted in partial protection against pneumonic plague challenge with 250 MLD Y. pestis CO92, immunization with recombinant F1 did not. rV10, a vaccine variant lacking LcrV residues 271-300, elicited protection against pneumonic plague, which seemed to be based on conformational antibodies directed against LcrV. Copyright © 2011 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

Comparative quantification of human intestinal bacteria based on cPCR and LDR/LCR

PubMed Central

Tang, Zhou-Rui; Li, Kai; Zhou, Yu-Xun; Xiao, Zhen-Xian; Xiao, Jun-Hua; Huang, Rui; Gu, Guo-Hao

2012-01-01

AIM: To establish a multiple detection method based on comparative polymerase chain reaction (cPCR) and ligase detection reaction (LDR)/ligase chain reaction (LCR) to quantify the intestinal bacterial components. METHODS: Comparative quantification of 16S rDNAs from different intestinal bacterial components was used to quantify multiple intestinal bacteria. The 16S rDNAs of different bacteria were amplified simultaneously by cPCR. The LDR/LCR was examined to actualize the genotyping and quantification. Two beneficial (Bifidobacterium, Lactobacillus) and three conditionally pathogenic bacteria (Enterococcus, Enterobacterium and Eubacterium) were used in this detection. With cloned standard bacterial 16S rDNAs, standard curves were prepared to validate the quantitative relations between the ratio of original concentrations of two templates and the ratio of the fluorescence signals of their final ligation products. The internal controls were added to monitor the whole detection flow. The quantity ratio between two bacteria was tested. RESULTS: cPCR and LDR revealed obvious linear correlations with standard DNAs, but cPCR and LCR did not. In the sample test, the distributions of the quantity ratio between each two bacterial species were obtained. There were significant differences among these distributions in the total samples. But these distributions of quantity ratio of each two bacteria remained stable among groups divided by age or sex. CONCLUSION: The detection method in this study can be used to conduct multiple intestinal bacteria genotyping and quantification, and to monitor the human intestinal health status as well. PMID:22294830

Comparative quantification of human intestinal bacteria based on cPCR and LDR/LCR.

PubMed

Tang, Zhou-Rui; Li, Kai; Zhou, Yu-Xun; Xiao, Zhen-Xian; Xiao, Jun-Hua; Huang, Rui; Gu, Guo-Hao

2012-01-21

To establish a multiple detection method based on comparative polymerase chain reaction (cPCR) and ligase detection reaction (LDR)/ligase chain reaction (LCR) to quantify the intestinal bacterial components. Comparative quantification of 16S rDNAs from different intestinal bacterial components was used to quantify multiple intestinal bacteria. The 16S rDNAs of different bacteria were amplified simultaneously by cPCR. The LDR/LCR was examined to actualize the genotyping and quantification. Two beneficial (Bifidobacterium, Lactobacillus) and three conditionally pathogenic bacteria (Enterococcus, Enterobacterium and Eubacterium) were used in this detection. With cloned standard bacterial 16S rDNAs, standard curves were prepared to validate the quantitative relations between the ratio of original concentrations of two templates and the ratio of the fluorescence signals of their final ligation products. The internal controls were added to monitor the whole detection flow. The quantity ratio between two bacteria was tested. cPCR and LDR revealed obvious linear correlations with standard DNAs, but cPCR and LCR did not. In the sample test, the distributions of the quantity ratio between each two bacterial species were obtained. There were significant differences among these distributions in the total samples. But these distributions of quantity ratio of each two bacteria remained stable among groups divided by age or sex. The detection method in this study can be used to conduct multiple intestinal bacteria genotyping and quantification, and to monitor the human intestinal health status as well.

The Transcription Factors TBX2 and TBX3 Interact with Human Papillomavirus 16 (HPV16) L2 and Repress the Long Control Region of HPVs

PubMed Central

Schneider, Marc A.; Scheffer, Konstanze D.; Bund, Timo; Boukhallouk, Fatima; Lambert, Carsten; Cotarelo, Cristina; Pflugfelder, Gert O.

2013-01-01

The minor capsid protein L2 of human papillomaviruses (HPVs) has multiple functions during the viral life cycle. Although L2 is required for effective invasion and morphogenesis, only a few cellular interaction partners are known so far. Using yeast two-hybrid screening, we identified the transcription factor TBX2 as a novel interaction partner of HPV type 16 (HPV16) L2. Coimmunoprecipitations and immunofluorescence analyses confirmed the L2-TBX2 interaction and revealed that L2 also interacts with TBX3, another member of the T-box family. Transcription of the early genes during HPV infection is under the control of an upstream enhancer and early promoter region, the long control region (LCR). In promoter-reporter gene assays, we observed that TBX2 and TBX3 repress transcription from the LCR and that this effect is enhanced by L2. Repression of the HPV LCR by TBX2/3 seems to be a conserved mechanism, as it was also observed with the LCRs of different HPV types. Finally, interaction of TBX2 with the LCR was detected by chromatin immunoprecipitation, and we found a strong colocalization of L2 and TBX2 in HPV16-positive cervical intraepithelial neoplasia (CIN) I-II tissue sections. These results suggest that TBX2/3 might play a role in the regulation of HPV gene expression during the viral life cycle. PMID:23388722

Mutation detection of E6 and LCR genes from HPV 16 associated with carcinogenesis.

PubMed

Mosmann, Jessica P; Monetti, Marina S; Frutos, Maria C; Kiguen, Ana X; Venezuela, Raul F; Cuffini, Cecilia G

2015-01-01

Human papillomavirus (HPV) is responsible for one of the most frequent sexually transmitted infections. The first phylogenetic analysis was based on a LCR region fragment. Nowadays, 4 variants are known: African (Af-1, Af-2), Asian-American (AA) and European (E). However the existence of sub-lineages of the European variant havs been proposed, specific mutations in the E6 and LCR sequences being possibly related to persistent viral infections. The aim of this study was a phylogenetic study of HPV16 sequences of endocervical samples from Cordoba, in order to detect the circulating lineages and analyze the presence of mutations that could be correlated with malignant disease. The phylogenetic analysis determined that 86% of the samples belonged to the E variant, 7% to AF-1 and the remaining 7% to AF-2. The most frequent mutation in LCR sequences was G7521A, in 80% of the analyzed samples; it affects the binding site of a transcription factor that could contribute to carcinogenesis. In the E6 sequences, the most common mutation was T350G (L83V), detected in 67% of the samples, associated with increased risk of persistent infection. The high detection rate of the European lineage correlated with patterns of human migration. This study emphasizes the importance of recognizing circulating lineages, as well as the detection of mutations associated with high-grade neoplastic lesions that could be correlated to the development of carcinogenic lesions.

Leanness and heightened nonresting energy expenditure: role of skeletal muscle activity thermogenesis

PubMed Central

Mukherjee, Sromona; Shukla, Charu; Britton, Steven L.; Koch, Lauren G.; Shi, Haifei; Novak, Colleen M.

2014-01-01

A high-calorie diet accompanied by low levels of physical activity (PA) accounts for the widespread prevalence of obesity today, and yet some people remain lean even in this obesogenic environment. Here, we investigate the cause for this exception. A key trait that predicts high PA in both humans and laboratory rodents is intrinsic aerobic capacity. Rats artificially selected as high-capacity runners (HCR) are lean and consistently more physically active than their low-capacity runner (LCR) counterparts; this applies to both males and females. Here, we demonstrate that HCR show heightened total energy expenditure (TEE) and hypothesize that this is due to higher nonresting energy expenditure (NREE; includes activity EE). After matching for body weight and lean mass, female HCR consistently had heightened nonresting EE, but not resting EE, compared with female LCR. Because of the dominant role of skeletal muscle in nonresting EE, we examined muscle energy use. We found that lean female HCR had higher muscle heat dissipation during activity, explaining their low economy of activity and high activity EE. This may be due to the amplified skeletal muscle expression levels of proteins involved in EE and reduced expression levels of proteins involved in energy conservation in HCR relative to LCR. This is also associated with an increased sympathetic drive to skeletal muscle in HCR compared with LCR. We find little support for the hypothesis that resting metabolic rate is correlated with maximal aerobic capacity if body size and composition are fully considered; rather, the critical factor appears to be activity thermogenesis. PMID:24398400

Electromobility Shift Assay Reveals Evidence in Favor of Allele-Specific Binding of RUNX1 to the 5' Hypersensitive Site 4-Locus Control Region.

PubMed

Dehghani, Hossein; Ghobakhloo, Sepideh; Neishabury, Maryam

2016-08-01

In our previous studies on the Iranian β-thalassemia (β-thal) patients, we identified an association between the severity of the β-thal phenotype and the polymorphic palindromic site at the 5' hypersensitive site 4-locus control region (5'HS4-LCR) of the β-globin gene cluster. Furthermore, a linkage disequilibrium was observed between this region and XmnI-HBG2 in the patient population. Based on this data, it was suggested that the well-recognized phenotype-ameliorating role assigned to positive XmnI could be associated with its linked elements in the LCR. To investigate the functional significance of polymorphisms at the 5'HS4-LCR, we studied its influence on binding of transcription factors. Web-based predictions of transcription factor binding revealed a binding site for runt-related transcription factor 1 (RUNX1), when the allele at the center of the palindrome (TGGGG(A/G)CCCCA) was A but not when it was G. Furthermore, electromobility shift assay (EMSA) presented evidence in support of allele-specific binding of RUNX1 to 5'HS4. Considering that RUNX1 is a well-known regulator of hematopoiesis, these preliminary data suggest the importance of further studies to confirm this interaction and consequently investigate its functional and phenotypical relevance. These studies could help us to understand the molecular mechanism behind the phenotype modifying role of the 5'HS4-LCR polymorphic palindromic region (rs16912979), which has been observed in previous studies.

CpG methylation of HPV 16 LCR at E2 binding site proximal to P97 is associated with cervical cancer in presence of intact E2.

PubMed

Bhattacharjee, Bornali; Sengupta, Sharmila

2006-10-25

Human papillomavirus type 16 (HPV-16) E2 protein negatively regulates transcription of the E6 and E7 genes. This study was done to test the hypothesis that methylation of the HPV 16 long control region (LCR) is overrepresented among cervical cancer (CaCx) cases compared to cytologically normal controls harboring intact E2 gene. Methylation of the E2 binding site (E2BS-I), proximal to the P97 promoter, was assessed by HpaII/ MspI restriction digestion while McrBC digestion was used to assess LCR-E6 (7289-540) for 57 CaCx samples and 15 normal controls. E2BS-I methylation was found to be significantly higher (56.14%) in cases compared to (20%) controls [OR(age-adjusted) (95% CI): 4.53 (1.05-19.43) p=0.042]. The difference between cases (54.39%) and controls (40%) with respect to LCR-E6 methylation status [OR(age-adjusted) (95% CI): 1.77(0.5-6.3); p=0.38] was not significant. Sequencing of a randomly selected set of 13 methylated malignant samples revealed absence or rare presence, of methylation at CpGs 7579, 7535, 7683 and 7862 respectively. Methylation was found to be more at CpGs within E2 binding sites proximal to the P97 promoter. These results indicate the involvement of E2 binding site methylation in presence of intact E2, leading to loss of E2 repressor activity in CaCx.

PMS2 inactivation by a complex rearrangement involving an HERV retroelement and the inverted 100-kb duplicon on 7p22.1.

PubMed

Vogt, Julia; Wernstedt, Annekatrin; Ripperger, Tim; Pabst, Brigitte; Zschocke, Johannes; Kratz, Christian; Wimmer, Katharina

2016-11-01

Biallelic PMS2 mutations are responsible for more than half of all cases of constitutional mismatch repair deficiency (CMMRD), a recessively inherited childhood cancer predisposition syndrome. The mismatch repair gene PMS2 is partly embedded within one copy of an inverted 100-kb low-copy repeat (LCR) on 7p22.1. In an individual with CMMRD syndrome, PMS2 was found to be homozygously inactivated by a complex chromosomal rearrangement, which separates the 5'-part from the 3'-part of the gene. The rearrangement involves sequences of the inverted 100-kb LCR and a human endogenous retrovirus element and may be associated with an inversion that is indistinguishable from the known inversion polymorphism affecting the ~0.7-Mb sequence intervening the LCR. Its formation is best explained by a replication-based mechanism (RBM) such as fork stalling and template switching/microhomology-mediated break-induced replication (FoSTeS/MMBIR). This finding supports the hypothesis that the inverted LCR can not only facilitate the formation of the non-allelic homologous recombination-mediated inversion polymorphism but it also promotes the occurrence of more complex rearrangements that can be associated with a large inversion, as well, but are mediated by a RBM. This further suggests that among the inversion polymorphism on 7p22.1, more complex rearrangements might be hidden. Furthermore, as the locus is embedded in a common fragile site (CFS) region, this rearrangement also supports the recently raised hypothesis that CFS sequence motifs may facilitate replication-based rearrangement mechanisms.

PMS2 inactivation by a complex rearrangement involving an HERV retroelement and the inverted 100-kb duplicon on 7p22.1

PubMed Central

Vogt, Julia; Wernstedt, Annekatrin; Ripperger, Tim; Pabst, Brigitte; Zschocke, Johannes; Kratz, Christian; Wimmer, Katharina

2016-01-01

Biallelic PMS2 mutations are responsible for more than half of all cases of constitutional mismatch repair deficiency (CMMRD), a recessively inherited childhood cancer predisposition syndrome. The mismatch repair gene PMS2 is partly embedded within one copy of an inverted 100-kb low-copy repeat (LCR) on 7p22.1. In an individual with CMMRD syndrome, PMS2 was found to be homozygously inactivated by a complex chromosomal rearrangement, which separates the 5′-part from the 3′-part of the gene. The rearrangement involves sequences of the inverted 100-kb LCR and a human endogenous retrovirus element and may be associated with an inversion that is indistinguishable from the known inversion polymorphism affecting the ~0.7-Mb sequence intervening the LCR. Its formation is best explained by a replication-based mechanism (RBM) such as fork stalling and template switching/microhomology-mediated break-induced replication (FoSTeS/MMBIR). This finding supports the hypothesis that the inverted LCR can not only facilitate the formation of the non-allelic homologous recombination-mediated inversion polymorphism but it also promotes the occurrence of more complex rearrangements that can be associated with a large inversion, as well, but are mediated by a RBM. This further suggests that among the inversion polymorphism on 7p22.1, more complex rearrangements might be hidden. Furthermore, as the locus is embedded in a common fragile site (CFS) region, this rearrangement also supports the recently raised hypothesis that CFS sequence motifs may facilitate replication-based rearrangement mechanisms. PMID:27329736

Nucleus accumbens cocaine-amphetamine regulated transcript mediates food intake during novelty conflict

PubMed Central

Burghardt, PR; Krolewski, DM; Dykhuis, KE; Ching, J; Pinawin, AM; Britton, SL; Koch, LG; Watson, SJ; Akil, H.

2016-01-01

Obesity is a persistent and pervasive problem, particularly in industrialized nations. It has come to be appreciated that the metabolic health of an individual can influence brain function and subsequent behavioral patterns. To examine the relationship between metabolic phenotype and central systems that regulate behavior, we tested rats with divergent metabolic phenotypes (Low Capacity Runner: LCR vs. High Capacity Runner: HCR) for behavioral responses to the conflict between hunger and environmental novelty using the novelty suppressed feeding (NSF) paradigm. Additionally, we measured expression of mRNA, for peptides involved in energy management, in response to fasting. Following a 24-h fast, LCR rats showed lower latencies to begin eating in a novel environment compared to HCR rats. A 48-h fast equilibrated the latency to begin eating in the novel environment. A 24-h fast differentially affected expression of cocaine-amphetamine regulated transcript (CART) mRNA in the nucleus accumbens (NAc), where 24-h of fasting reduced CART mRNA in LCR rats. Bilateral microinjections of CART 55–102 peptide into the NAc increased the latency to begin eating in the NSF paradigm following a 24-h fast in LCR rats. These results indicate that metabolic phenotype influences how animals cope with the conflict between hunger and novelty, and that these differences are at least partially mediated by CART signaling in the NAc. For individuals with poor metabolic health who have to navigate food-rich and stressful environments, changes in central systems that mediate conflicting drives may feed into the rates of obesity and exacerbate the difficulty individuals have in maintaining weight loss. PMID:26926827

Detection of hypermutated human papillomavirus type 16 genome by Next-Generation Sequencing.

PubMed

Wakae, Kousho; Aoyama, Satoru; Wang, Zhe; Kitamura, Kouichi; Liu, Guangyan; Monjurul, Ahasan Md; Koura, Miki; Imayasu, Mieko; Sakamoto, Naoya; Nakamura, Mitsuhiro; Kyo, Satoru; Kondo, Satoru; Fujiwara, Hiroshi; Yoshizaki, Tomokazu; Kukimoto, Iwao; Yamaguchi, Katsushi; Shigenobu, Shuji; Nishiyama, Tomoaki; Muramatsu, Masamichi

2015-11-01

Human papillomavirus type 16 (HPV16) is a major cause of cervical cancer. We previously demonstrated that C-to-T and G-to-A hypermutations accumulated in the HPV16 genome by APOBEC3 expression in vitro. To investigate in vivo characteristics of hypermutation, differential DNA denaturation-PCR (3D-PCR) was performed using three clinical specimens obtained from HPV16-positive cervical dysplasia, and detected hypermutation from two out of three specimens. One sample accumulating hypermutations in both E2 and the long control region (LCR) was further subjected to Next-Generation Sequencing, revealing that hypermutations spread across the LCR and all early genes. Notably, hypermutation was more frequently observed in the LCR, which contains a viral replication origin and the early promoter. APOBEC3 expressed abundantly in an HPV16-positive cervix, suggesting that single-stranded DNA exposed during viral replication and transcription may be efficient targets for deamination. The results further strengthen a role of APOBEC3 in introducing HPV16 hypermutation in vivo. Copyright © 2015 Elsevier Inc. All rights reserved.

Non-native fish control below Glen Canyon Dam - Report from a structured decision-making project

USGS Publications Warehouse

Runge, Michael C.; Bean, Ellen; Smith, David; Kokos, Sonja

2011-01-01

This report describes the results of a structured decision-making project by the U.S. Geological Survey to provide substantive input to the Bureau of Reclamation (Reclamation) for use in the preparation of an Environmental Assessment concerning control of non-native fish below Glen Canyon Dam. A forum was created to allow the diverse cooperating agencies and Tribes to discuss, expand, and articulate their respective values; to develop and evaluate a broad set of potential control alternatives using the best available science; and to define individual preferences of each group on how to manage the inherent trade-offs in this non-native fish control problem. This project consisted of two face-to-face workshops, held in Mesa, Arizona, October 18-20 and November 8-10, 2010. At the first workshop, a diverse set of objectives was discussed, which represented the range of concerns of those agencies and Tribes present. A set of non-native fish control alternatives ('hybrid portfolios') was also developed. Over the 2-week period between the two workshops, four assessment teams worked to evaluate the control alternatives against the array of objectives. At the second workshop, the results of the assessment teams were presented. Multi-criteria decision analysis methods were used to examine the trade-offs inherent in the problem, and allowed the participating agencies and Tribes to express their individual judgments about how those trade-offs should best be managed in Reclamation`s selection of a preferred alternative. A broad array of objectives was identified and defined, and an effort was made to understand how these objectives are likely to be achieved by a variety of strategies. In general, the objectives reflected desired future conditions over 30 years. A rich set of alternative approaches was developed, and the complex structure of those alternatives was documented. Multi-criteria decision analysis methods allowed the evaluation of those alternatives against the array of objectives, with the values of individual agencies and tribes deliberately preserved. Trout removal strategies aimed at the Paria to Badger Rapid reach (PBR), with a variety of permutations in deference to cultural values, and with backup removal at the Little Colorado River reach (LCR) if necessary, were identified as top-ranking portfolios for all agencies and Tribes. These PBR/LCR removal portfolios outperformed LCR-only removal portfolios, for cultural reasons and for effectiveness - the probability of keeping the humpback chub population above a desired threshold was estimated to be higher under the PBR/LCR portfolios than the LCR-only portfolios. The PBR/LCR removal portfolios also outperformed portfolios based on flow manipulations, primarily because of the effect of sport fishery and wilderness recreation objectives, as well as cultural objectives. The preference for the PBR/LCR removal portfolios was quite robust to variation in the objective weights and to uncertainty about the underlying dynamics, at least over the ranges of uncertainty investigated. Examination of the effect of uncertainty on the recommended outcomes allowed us to complete a 'value of information' analysis. The results of this analysis led to an adaptive strategy that includes three possible long-term management actions (no action; LCR removal; or PBR removal) and seeks to reduce uncertainty about the following two issues: the degree to which rainbow trout limit chub populations, and the effectiveness of PBR removal to reduce trout emigration downstream into Marble and eastern Grand Canyons, where the largest population of humpback chub exist. In the face of uncertainty about the effectiveness of PBR removal, a case might be made for including flow manipulations in an adaptive strategy, but formal analysis of this case was not conducted. The full set of conclusions described above is not definitive, however. This analysis described in this report is a simplified depiction of the t

Investigation of photoluminescence and dielectric properties of pure and Fe doped nickel oxide nanoparticles

NASA Astrophysics Data System (ADS)

Gupta, Jhalak; Ahmad, Arham S.

2018-05-01

The nanocrystallites of pure and Fe doped Nickel Oxide (NiO) were synthesized by the cost effective co-precipitation method using nickel nitrate as the initial precursor. The synthesized nickel oxide nanoparticles were characterized by X-Ray Diffraction (XRD), Photoluminiscence Spectroscopy (PL), LCR meter. The crystallite size of synthesized pure Nickel Oxide nanoparticles obtained by XRD using Debye Scherer's formula was found to be 21.8nm and the size decreases on increasing the dopant concentration. The optical properties were analyzed by PL and dielectric ones by using LCR meter.

DNASynth: a software application to optimization of artificial gene synthesis

NASA Astrophysics Data System (ADS)

Muczyński, Jan; Nowak, Robert M.

2017-08-01

DNASynth is a client-server software application in which the client runs in a web browser. The aim of this program is to support and optimize process of artificial gene synthesizing using Ligase Chain Reaction. Thanks to LCR it is possible to obtain DNA strand coding defined by user peptide. The DNA sequence is calculated by optimization algorithm that consider optimal codon usage, minimal energy of secondary structures and minimal number of required LCR. Additionally absence of sequences characteristic for defined by user set of restriction enzymes is guaranteed. The presented software was tested on synthetic and real data.

Ex vivo investigations of laser auricular cartilage reshaping with carbon dioxide spray cooling in a rabbit model

PubMed Central

Wu, Edward C.; Sun, Victor; Manuel, Cyrus T.; Protsenko, Dmitriy E.; Jia, Wangcun; Nelson, J. Stuart; Wong, Brian J. F.

2014-01-01

Laser cartilage reshaping (LCR) with cryogen spray cooling is a promising modality for producing cartilage shape change while reducing cutaneous thermal injury. However, LCR in thicker tissues, such as auricular cartilage, requires higher laser power, thus increasing cooling requirements. To eliminate the risks of freeze injury characteristic of high cryogen spray pulse rates, a carbon dioxide (CO2) spray, which evaporates rapidly from the skin, has been proposed as the cooling medium. This study aims to identify parameter sets which produce clinically significant reshaping while producing minimal skin thermal injury in LCR with CO2 spray cooling in ex vivo rabbit auricular cartilage. Excised whole rabbit ears were mechanically deformed around a cylindrical jig and irradiated with a 1.45-μm wavelength diode laser (fluence 12–14 J/cm2 per pulse, four to six pulse cycles per irradiation site, five to six irradiation sites per row for four rows on each sample) with concomitant application of CO2 spray (pulse duration 33–85 ms) to the skin surface. Bend angle measurements were performed before and after irradiation, and the change quantified. Surface temperature distributions were measured during irradiation/cooling. Maximum skin surface temperature ranged between 49.0 to 97.6 °C following four heating/cooling cycles. Significant reshaping was achieved with all laser dosimetry values with a 50–70 °C difference noted between controls (no cooling) and irradiated ears. Increasing cooling pulse duration yielded progressively improved gross skin protection during irradiation. CO2 spray cooling may potentially serve as an alternative to traditional cryogen spray cooling in LCR and may be the preferred cooling medium for thicker tissues. Future studies evaluating preclinical efficacy in an in vivo rabbit model are in progress. PMID:23307439

Enhancer‐promoter Activity of Human Papillomavirus Type 16 Long Control Regions Isolated from Cell Lines SiHa and CaSki and Cervical Cancer Biopsies

PubMed Central

Kozuka, Takuyo; Aoki, Yukimasa; Nakagawa, Keiichi; Ohtomo, Kuni; Yoshikawa, Hiroyuki; Matsumoto, Koji; Yoshiike, Kunito

2000-01-01

Expression of human papillomavirus 16 (HPV‐16) oncogenes is markedly higher in cervical cancer cells than in precancerous cells, and the elevated expression is believed to be required for the malignant phenotypes. We compared cancer cell lines CaSki (with 200 to 400 copies of HPV‐16 DNA per cell) and SiHa (with one to two copies of HPV‐16 DNA per cell) for the E7 expression in cells and the enhancer‐promoter activity of the isolated viral long control region (LCR). Although these parameters per cell were 10‐fold higher in CaSki than in SiHa, the levels of the E7 mRNA and protein per HPV DNA copy were 10‐ to 20‐fold higher in SiHa than in CaSki. Characterization of the isolated LCRs showed that, whereas the LCR from CaSki resembled the prototype in structure and activity, the LCR from SiHa, with a deletion of 38 base pairs, enhanced transcription from P97 as assayed by using a plasmid capable of expressing luciferase. The upregulation appeared to be due to removal of one of the silencer YY1‐binding sites. Furthermore, we isolated and characterized LCRs from 51 cervical cancer patients’ biopsies. Among them, one with a deletion including YY1‐binding sites and the other with a substitution in a YY1‐motif were found to enhance the transcription. These findings suggest that mutation affecting YY1‐motifs in the LCR is one of the mechanisms enhancing the viral oncogene expression in the course of progression of cancer cells. PMID:10760685

Conformational stability and differential structural analysis of LcrV, PcrV, BipD, and SipD from type III secretion systems

PubMed Central

Espina, Marianela; Ausar, S. Fernando; Middaugh, C. Russell; Baxter, M. Aaron; Picking, William D.; Picking, Wendy L.

2007-01-01

Diverse Gram-negative bacteria use type III secretion systems (T3SS) to translocate effector proteins into the cytoplasm of eukaryotic cells. The type III secretion apparatus (T3SA) consists of a basal body spanning both bacterial membranes and an external needle. A sensor protein lies at the needle tip to detect environmental signals that trigger type III secretion. The Shigella flexneri T3SA needle tip protein, invasion plasmid antigen D (IpaD), possesses two independently folding domains in vitro. In this study, the solution behavior and thermal unfolding properties of IpaD's functional homologs SipD (Salmonella spp.), BipD (Burkholderia pseudomallei), LcrV (Yersinia spp.), and PcrV (Pseudomonas aeruginosa) were examined to identify common features within this protein family. CD and FTIR data indicate that all members within this group are α-helical with properties consistent with an intramolecular coiled-coil. SipD showed the most complex unfolding profile consisting of two thermal transitions, suggesting the presence of two independently folding domains. No evidence of multiple folding domains was seen, however, for BipD, LcrV, or PcrV. Thermal studies, including DSC, revealed significant destabilization of LcrV, PcrV, and BipD after N-terminal deletions. This contrasted with SipD and IpaD, which behaved like two-domain proteins. The results suggest that needle tip proteins share significant core structural similarity and thermal stability that may be the basis for their common function. Moreover, IpaD and SipD possess properties that distinguish them from the other tip proteins. PMID:17327391

A Bivalent Typhoid Live Vector Vaccine Expressing both Chromosome- and Plasmid-Encoded Yersinia pestis Antigens Fully Protects against Murine Lethal Pulmonary Plague Infection

PubMed Central

Wang, Jin Yuan; Carrasco, Jose A.; Lloyd, Scott A.; Mellado-Sanchez, Gabriela; Diaz-McNair, Jovita; Franco, Olga; Buskirk, Amanda D.; Nataro, James P.; Pasetti, Marcela F.

2014-01-01

Live attenuated bacteria hold great promise as multivalent mucosal vaccines against a variety of pathogens. A major challenge of this approach has been the successful delivery of sufficient amounts of vaccine antigens to adequately prime the immune system without overattenuating the live vaccine. Here we used a live attenuated Salmonella enterica serovar Typhi strain to create a bivalent mucosal plague vaccine that produces both the protective F1 capsular antigen of Yersinia pestis and the LcrV protein required for secretion of virulence effector proteins. To reduce the metabolic burden associated with the coexpression of F1 and LcrV within the live vector, we balanced expression of both antigens by combining plasmid-based expression of F1 with chromosomal expression of LcrV from three independent loci. The immunogenicity and protective efficacy of this novel vaccine were assessed in mice by using a heterologous prime-boost immunization strategy and compared to those of a conventional strain in which F1 and LcrV were expressed from a single low-copy-number plasmid. The serum antibody responses to lipopolysaccharide (LPS) induced by the optimized bivalent vaccine were indistinguishable from those elicited by the parent strain, suggesting an adequate immunogenic capacity maintained through preservation of bacterial fitness; in contrast, LPS titers were 10-fold lower in mice immunized with the conventional vaccine strain. Importantly, mice receiving the optimized bivalent vaccine were fully protected against lethal pulmonary challenge. These results demonstrate the feasibility of distributing foreign antigen expression across both chromosomal and plasmid locations within a single vaccine organism for induction of protective immunity. PMID:25332120

Human papillomavirus type 56 polymorphism in Canadian women with and without cervical lesions.

PubMed

Rodrigues-Coutlée, Catherine; Archambault, Jacques; Money, Deborah; Ramanakumar, Agnihotram V; Raboud, Janet; Hankins, Catherine; Koushik, Anita; Richardson, Harriet; Brassard, Paul; Franco, Eduardo L; Coutlée, Francois

2013-12-01

The genomic diversity of high-risk human papillomaviruses (HPV) has been associated with viral persistence and HPV-induced lesions. Studies on HPV56 persistence are still pending. To assess the association between HPV56 polymorphism and HPV56 persistence and presence of high-grade cervical intraepithelial neoplasia (CIN2,3) or cancer. HPV56-positive cervical specimens from 204 women selected from a total of 4669 participants recruited in 5 epidemiological studies (parent studies) were further analyzed by PCR-sequencing of the long control region (LCR). Of the 81 women followed prospectively in cohort studies who could be classified, 34 had persistent and 47 had transient HPV56 infections. Variant HPV56-LCR-MTL-21 was detected more frequently in persistent infections (52.9%, 95% CI: 36.7-68.6%) than in transient infections (25.5%, 95% CI: 15.1-39.4). Considering only women recruited in a cohort of women infected or at high risk for HIV infection, infection with variant HPV56-LCR-MTL-21 (OR=4.4, 95% CI: 1.3-14.5) was significantly associated with HPV56 persistence controlling in multivariate analysis for high risk HPV detection and HIV infection. A variation at nucleotide 7800 in HPV56-LCR-MTL-21 resulted in the loss of a binding site for Elf-1 embedded in one of the E2 binding sites, a potential activator or repressor of expression of the HPV genome. HPV56 polymorphism was not associated with CIN2,3 or cancer in women enrolled in cross-sectional and case-control studies. Polymorphism in HPV56 may influence the risk that infections with this type will persist. Copyright © 2013 Elsevier B.V. All rights reserved.

Parasites of native and nonnative fishes of the Little Colorado River, Grand Canyon, Arizona

USGS Publications Warehouse

Choudhury, A.; Hoffnagle, T.L.; Cole, Rebecca A.

2004-01-01

A 2-yr, seasonal, parasitological study of 1,435 fish, belonging to 4 species of native fishes and 7 species of nonnative fishes from the lower Little Colorado River (LCR) and tributary creeks, Grand Canyon, Arizona, yielded 17 species of parasites. These comprised 1 myxozoan (Henneguya exilis), 2 copepods (Ergasilus arthrosis and Lernaea cyprinacea), 1 acarine (Oribatida gen. sp.), 1 piscicolid leech (Myzobdella lugubris), 4 monogeneans (Gyrodactylus hoffmani, Gyrodactylus sp., Dactylogyrus extensus, and Ligictaluridus floridanus), 4 nematodes (Contracaecum sp., Eustrongylides sp., Rhabdochona sp., and Truttaedacnitis truttae), 3 cestodes (Bothriocephalus acheilognathi, Corallobothrium fimbriatum, and Megathylacoides giganteum), and 2 trematodes (Ornithodiplostomum sp. and Posthodiplostomum sp.). Rhabdochona sp. was the only adult parasite native to the LCR. Infection intensities of Ornithodiplostomum sp. and B. acheilognathi were positively correlated with length of the humpback chub Gila cypha. Adult helminths showed a high degree of host specificity, except B. acheilognathi, which was recovered from all fish species examined but was most abundant in cyprinids. Abundance of B. acheilognathi in the humpback chub was highest in the fall and lowest in the summer in both reaches of the LCR. There was no major taxonomic difference in parasite assemblages between the 2 different reaches of the river (LC1 and LC2). Parasite community diversity was very similar in humpback chub, regardless of sampling site or time. The parasite fauna of the LCR is numerically dominated by B. acheilognathi and metacercariae of Ornithodiplostomum sp. The richest and most diverse component community occurred in a nonnative species, the channel catfish Ictalurus punctatus, but infracommunity species richness was highest in a native host, humpback chub.

Human Papillomavirus Type 16 Genetic Variants: Phylogeny and Classification Based on E6 and LCR

PubMed Central

Gheit, Tarik; Franceschi, Silvia; Vignat, Jerome; Burk, Robert D.; Sylla, Bakary S.; Tommasino, Massimo; Clifford, Gary M.

2012-01-01

Naturally occurring genetic variants of human papillomavirus type 16 (HPV16) are common and have previously been classified into 4 major lineages; European-Asian (EAS), including the sublineages European (EUR) and Asian (As), African 1 (AFR1), African 2 (AFR2), and North-American/Asian-American (NA/AA). We aimed to improve the classification of HPV16 variant lineages by using a large resource of HPV16-positive cervical samples collected from geographically diverse populations in studies on HPV and/or cervical cancer undertaken by the International Agency for Research on Cancer. In total, we sequenced the entire E6 genes and long control regions (LCRs) of 953 HPV16 isolates from 27 different countries worldwide. Phylogenetic analyses confirmed previously described variant lineages and subclassifications. We characterized two new sublineages within each of the lineages AFR1 and AFR2 that are robustly classified using E6 and/or the LCR. We could differentiate previously identified AA1, AA2, and NA sublineages, although they could not be distinguished by E6 alone, requiring the LCR for correct phylogenetic classification. We thus provide a classification system for HPV16 genomes based on 13 and 32 phylogenetically distinguishing positions in E6 and the LCR, respectively, that distinguish nine HPV16 variant sublineages (EUR, As, AFR1a, AFR1b, AFR2a, AFR2b, NA, AA1, and AA2). Ninety-seven percent of all 953 samples fitted this classification perfectly. Other positions were frequently polymorphic within one or more lineages but did not define phylogenetic subgroups. Such a standardized classification of HPV16 variants is important for future epidemiological and biological studies of the carcinogenic potential of HPV16 variant lineages. PMID:22491459

Deletion of the human beta-globin LCR 5'HS4 or 5'HS1 differentially affects beta-like globin gene expression in beta-YAC transgenic mice.

PubMed

Fedosyuk, Halyna; Peterson, Kenneth R

2007-01-01

A 213 kb human beta-globin locus yeast artificial chromosome (beta-YAC) was modified by homologous recombination to delete 2.9 kb of cross-species conserved sequence similarity encompassing the LCR 5' hypersensitive site (HS) 4 (Delta5'HS4 beta-YAC). In three transgenic mouse lines, completion of the gamma- to beta-globin switch during definitive erythropoiesis was delayed relative to wild-type beta-YAC mice. In addition, quantitative per-copy human beta-like globin mRNA levels were similar to wild-type beta-YAC transgenic lines, although beta-globin gene expression was slightly decreased in the day 12 fetal liver of Delta5'HS4 beta-YAC mice. A 0.8 kb 5'HS1 fragment was similarly deleted in the YAC. Three Delta5'HS1 beta-YAC transgenic lines were established. epsilon-globin gene expression was markedly reduced, approximately 16 fold, during primitive erythropoiesis compared to wild-type beta-YAC mice, but gamma-globin expression levels were unaffected. However, during the fetal stage of definitive erythropoiesis, gamma-globin gene expression was decreased approximately 4 fold at day 12 and approximately 5 fold at day 14. Temporal developmental expression profiles of the beta-like globin genes were unaffected by deletion of 5'HS1. Decreased expression of the epsilon- and gamma-globin genes is the first phenotype ascribed to a 5'HS1 mutation in the human beta-globin locus, suggesting that this HS does indeed have a role in LCR function beyond simply a combined synergism with the other LCR HSs.

HPV16 E2 gene disruption and polymorphisms of E2 and LCR: some significant associations with cervical cancer in Indian women.

PubMed

Bhattacharjee, Bornali; Sengupta, Sharmila

2006-02-01

We evaluated the status of the HPV16 E2 gene (disrupted or intact), nucleotide sequence alterations within intact E2 genes and LCR of HPV16 isolates in a group of CaCx cases (invasive squamous cell carcinomas, n = 81) and population controls (normal cervical scrapes, n = 27) from Indian women. E2 disruption was detected by amplifying the entire E2 gene with single set of primers, while overlapping primers were used to determine if any particular region got selectively disrupted. Nucleotide variations in E2 and LCR were analyzed by PCR amplification followed by bi-directional sequencing. The associations between the viral factors and CaCx were analyzed using Fisher's Exact or Chi-squared test and interpreted as OR (95% CI) and P values. E2 disruption was significantly higher among the cases [3.38 (1.07-10.72); P = 0.02], which was maximum in the region between nucleotides 3650 and 3872 (DNA-binding region). The European (E) variant was found to be the prevalent subgroup (87.76% among cases and 96.30% among the controls), and the remaining samples were Asian-American variants. Among the E subgroup, variation at position 7450 (T > C) within the E2-binding site-IV was found to be significantly higher among the E2 undisrupted cases (21/37; 56.76%), compared to controls (5/18; 27.78%) [3.41 (1.01-11.55); P = 0.03]. Besides HPV16 E2 disruption, LCR 7450T > C variation within undisrupted E2 of E subgroup appears to be a major factor contributing to the risk of CaCx development in Indian women. Furthermore, polymorphisms in the E2 gene of HPV16 may not be significant for disease risk.

Human papillomavirus type 16 genetic variants: phylogeny and classification based on E6 and LCR.

PubMed

Cornet, Iris; Gheit, Tarik; Franceschi, Silvia; Vignat, Jerome; Burk, Robert D; Sylla, Bakary S; Tommasino, Massimo; Clifford, Gary M

2012-06-01

Naturally occurring genetic variants of human papillomavirus type 16 (HPV16) are common and have previously been classified into 4 major lineages; European-Asian (EAS), including the sublineages European (EUR) and Asian (As), African 1 (AFR1), African 2 (AFR2), and North-American/Asian-American (NA/AA). We aimed to improve the classification of HPV16 variant lineages by using a large resource of HPV16-positive cervical samples collected from geographically diverse populations in studies on HPV and/or cervical cancer undertaken by the International Agency for Research on Cancer. In total, we sequenced the entire E6 genes and long control regions (LCRs) of 953 HPV16 isolates from 27 different countries worldwide. Phylogenetic analyses confirmed previously described variant lineages and subclassifications. We characterized two new sublineages within each of the lineages AFR1 and AFR2 that are robustly classified using E6 and/or the LCR. We could differentiate previously identified AA1, AA2, and NA sublineages, although they could not be distinguished by E6 alone, requiring the LCR for correct phylogenetic classification. We thus provide a classification system for HPV16 genomes based on 13 and 32 phylogenetically distinguishing positions in E6 and the LCR, respectively, that distinguish nine HPV16 variant sublineages (EUR, As, AFR1a, AFR1b, AFR2a, AFR2b, NA, AA1, and AA2). Ninety-seven percent of all 953 samples fitted this classification perfectly. Other positions were frequently polymorphic within one or more lineages but did not define phylogenetic subgroups. Such a standardized classification of HPV16 variants is important for future epidemiological and biological studies of the carcinogenic potential of HPV16 variant lineages.

Analyzing 5'HS3 and 5'HS4 LCR core regions and NF-E2 in Iranian thalassemia intermedia patients with normal or carrier status for beta-globin mutations.

PubMed

Neishabury, Maryam; Azarkeivan, Azita; Oberkanins, Christian; Abedini, Seyedeh Sedigheh; Zamani, Shahbaz; Najmabadi, Hossein

2011-03-15

Our data on 114 Iranian individuals with thalassemia intermedia phenotype revealed homozygous or compound heterozygous beta-globin mutations to be the predominant disease factor in 86.2% of cases. However, 8.2% of these individuals were found to be heterozygous or wild type for beta-globin mutations. In search for determinants outside of the beta-globin gene, which could be responsible for the unexpected thalassemia intermedia phenotype in these subjects, we screened the alpha-globin genes, the 5'HS3 and 5'HS4 regions of the beta-globin LCR, and the NF-E2 transcription factor for sequence variations in selected individuals. The -3.7 deletion was the only alpha-globin mutation detected, and no alterations were found in 5'HS3 and NF-E2. Sequence analysis of the 5'HS4 LCR core region identified three known SNPs in a single patient, who required irregular blood transfusions. The A/G polymorphism in the 5'HS4 palindromic region was also observed to be variable. Family studies were carried out on a female G/G homozygous patient, who received irregular blood transfusions. Her father, who had the same heterozygous IVSII-1 beta-globin mutation but the A/G genotype at the 5'HS4 palindromic site, presented with mild anemia and no requirement for blood transfusions. This suggests an impact of SNPs in the 5'HS4 LCR core region on the thalassemia phenotype and offers an interesting subject for further investigations in the Iranian population. Copyright © 2010 Elsevier Inc. All rights reserved.

Incorporating DNA barcodes into a multi-year inventory of the fishes of the hyperdiverse Lower Congo River, with a multi-gene performance assessment of the genus Labeo as a case study.

PubMed

Lowenstein, Jacob H; Osmundson, Todd W; Becker, Sven; Hanner, Robert; Stiassny, Melanie L J

2011-10-01

Here we describe preliminary efforts to integrate DNA barcoding into an ongoing inventory of the Lower Congo River (LCR) ichthyofauna. The 350 km stretch of the LCR from Pool Malebo to Boma includes the world's largest river rapids. The LCR ichthyofauna is hyperdiverse and rich in endemism due to high habitat heterogeneity, numerous dispersal barriers, and its downstream location in the basin. We have documented 328 species from the LCR, 25% of which are thought to be endemic. In addition to detailing progress made to generate a reference sequence library of DNA barcodes for these fishes, we ask how DNA can be used at the current stage of the Fish Barcode of Life initiative, as a work in progress currently of limited utility to a wide audience. Two possibilities that we explore are the potential for DNA barcodes to generate discrete diagnostic characters for species, and to help resolve problematic taxa lacking clear morphologically diagnostic characters such as many species of the cyprinid genus Labeo, which we use as a case study. Our molecular analysis helped to clarify the validity of some species that were the subject of historical debate, and we were able to construct a molecular key for all monophyletic and morphologically recognizable species. Several species sampled from across the Congo Basin and widely distributed throughout Central and West Africa were recovered as paraphyletic based on our molecular data. Our study underscores the importance of generating reference barcodes for specimens collected from, or in close proximity to, type localities, particularly where species are poorly understood taxonomically and the extent of their geographical distributions have yet to be established.

Ex vivo investigations of laser auricular cartilage reshaping with carbon dioxide spray cooling in a rabbit model.

PubMed

Wu, Edward C; Sun, Victor; Manuel, Cyrus T; Protsenko, Dmitriy E; Jia, Wangcun; Nelson, J Stuart; Wong, Brian J F

2013-11-01

Laser cartilage reshaping (LCR) with cryogen spray cooling is a promising modality for producing cartilage shape change while reducing cutaneous thermal injury. However, LCR in thicker tissues, such as auricular cartilage, requires higher laser power, thus increasing cooling requirements. To eliminate the risks of freeze injury characteristic of high cryogen spray pulse rates, a carbon dioxide (CO2) spray, which evaporates rapidly from the skin, has been proposed as the cooling medium. This study aims to identify parameter sets which produce clinically significant reshaping while producing minimal skin thermal injury in LCR with CO2 spray cooling in ex vivo rabbit auricular cartilage. Excised whole rabbit ears were mechanically deformed around a cylindrical jig and irradiated with a 1.45-μm wavelength diode laser (fluence 12-14 J/cm(2) per pulse, four to six pulse cycles per irradiation site, five to six irradiation sites per row for four rows on each sample) with concomitant application of CO2 spray (pulse duration 33-85 ms) to the skin surface. Bend angle measurements were performed before and after irradiation, and the change quantified. Surface temperature distributions were measured during irradiation/cooling. Maximum skin surface temperature ranged between 49.0 to 97.6 °C following four heating/cooling cycles. Significant reshaping was achieved with all laser dosimetry values with a 50-70 °C difference noted between controls (no cooling) and irradiated ears. Increasing cooling pulse duration yielded progressively improved gross skin protection during irradiation. CO2 spray cooling may potentially serve as an alternative to traditional cryogen spray cooling in LCR and may be the preferred cooling medium for thicker tissues. Future studies evaluating preclinical efficacy in an in vivo rabbit model are in progress.

Nondestructive Testing Residual Stress Using Ultrasonic Critical Refracted Longitudinal Wave

NASA Astrophysics Data System (ADS)

Xu, Chunguang; Song, Wentao; Pan, Qinxue; Li, Huanxin; Liu, Shuai

Residual stress has significant impacts on the performance of the mechanical components, especially on its strength, fatigue life and corrosion resistance and dimensional stability. Based on theory of acoustoelasticity, the testing principle of ultrasonic LCR wave method is analyzed. The testing system of residual stress is build. The method of calibration of stress coefficient is proposed in order to improve the detection precision. At last, through experiments and applications on residual stress testing of oil pipeline weld joint, vehicle's torsion shaft, glass and ceramics, gear tooth root, and so on, the result show that it deserved to be studied deeply on application and popularization of ultrasonic LCR wave method.

Plague vaccines and the molecular basis of immunity against Yersinia pestis.

PubMed

Quenee, Lauriane E; Schneewind, Olaf

2009-12-01

Yersinia pestis is the causative agent of bubonic and pneumonic plague, human diseases with high mortality. Due to the microbe's ability to spread rapidly, plague epidemics present a serious public health threat. A search for prophylactic measures was initially based on historical reports of bubonic plague survivors and their apparent immunity. Due to safety and efficacy concerns, killed whole-cell preparations or live-attenuated plague vaccines are no longer considered in the United States. Vaccine developers have focused on specific subunits of plague bacteria. LcrV, a protein at the tip of type III secretion needles, and F1, the capsular pilus antigen, are both recognized as plague protective antigens. Antibodies against LcrV and F1 interfere with Y. pestis type III injection of host cells. While LcrV is absolutely essential for Y. pestis virulence, expression of F1 is dispensable for plague pathogenesis in small animals, non-human primates and presumably also in humans. Several subunit vaccines, for example rF1+rV (rYP002), rF1V or rV10, are being developed to generate plague protection in humans. Efficacy testing and licensure for human use requires the establishment of correlates for plague immunity.

Neonatal mucosal immunization with a non-living, non-genetically modified Lactococcus lactis vaccine carrier induces systemic and local Th1-type immunity and protects against lethal bacterial infection

PubMed Central

Ramirez, Karina; Ditamo, Yanina; Rodriguez, Liliana; Picking, Wendy L.; van Roosmalen, Maarten L.; Leenhouts, Kees; Pasetti, Marcela F.

2010-01-01

Safe and effective immunization of newborns and infants can significantly reduce childhood mortality, yet conventional vaccines have been largely unsuccessful in stimulating the neonatal immune system. We explored the capacity of a novel mucosal antigen delivery system consisting of non-living, non-genetically modified Lactococcus lactis particles, designated Gram-positive Enhancer Matrix (GEM), to induce immune responses in the neonatal setting. Yersinia pestis LcrV, used as model protective antigen, was displayed on the GEM particles. Newborn mice immunized intranasally with GEM-LcrV developed LcrV-specific antibodies, Th1-type cell-mediated immunity, and were protected against lethal Y. pestis (plague) infection. The GEM particles activated and enhanced the maturation of neonatal dendritic cells both in vivo and in vitro. These dendritic cells showed increased capacities for secretion of pro-inflammatory and Th1-cell polarizing cytokines, antigen presentation and stimulation of CD4+ and CD8+ T cells. These data show that mucosal immunization with L. lactis GEM particles carrying vaccine antigens represents a promising approach to prevent infectious diseases early in life. PMID:19924118

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

Di Prinzio, Renato; Almeida, Carlos Eduardo de; Laboratorio de Ciencias Radiologicas-Universidade do Estado do Rio de Janeiro

In Brazil there are over 100 high dose rate (HDR) brachytherapy facilities using well-type chambers for the determination of the air kerma rate of {sup 192}Ir sources. This paper presents the methodology developed and extensively tested by the Laboratorio de Ciencias Radiologicas (LCR) and presently in use to calibrate those types of chambers. The system was initially used to calibrate six well-type chambers of brachytherapy services, and the maximum deviation of only 1.0% was observed between the calibration coefficients obtained and the ones in the calibration certificate provided by the UWADCL. In addition to its traceability to the Brazilian Nationalmore » Standards, the whole system was taken to University of Wisconsin Accredited Dosimetry Calibration Laboratory (UWADCL) for a direct comparison and the same formalism to calculate the air kerma was used. The comparison results between the two laboratories show an agreement of 0.9% for the calibration coefficients. Three Brazilian well-type chambers were calibrated at the UWADCL, and by LCR, in Brazil, using the developed system and a clinical HDR machine. The results of the calibration of three well chambers have shown an agreement better than 1.0%. Uncertainty analyses involving the measurements made both at the UWADCL and LCR laboratories are discussed.« less

Analysis of Critically Refracted Longitudinal and Lamb Waves for Stress Characterization

NASA Astrophysics Data System (ADS)

Pei, Ning

The global production of metal, in particular, steel and aluminum keeps increasing. This material is used with various fabrication processes, such as, welding, forging, and rolling that can induce stresses in the material that can subsequently impact product performance and cause phenomena such as cracking and corrosion. When investigating plate materials it is necessary to map both texture and stress under a range of loading conditions. To address these needs a wide range of both destructive and nondestructive tools have been used. One family of methods are those based on ultrasonic measurements that relate ultrasonic velocity to properties, in particular stress. Two particular challenges are faced which are the relative insensitivity of compression and shear waves to stress and that there are also other factors which can also change velocity and these are temperature, texture and grain size. This project focused on an analysis of ultrasonic velocity measurements and specifically ways to improve performance and capabilities for stress characterization. Two approaches were considered and are reported: the critically refracted ultrasonic longitudinal (LCR) wave and higher order Lamb waves. The LCR wave method was modelled and optimized based on the fact that the sensitivity between waves and stress can reach maximum when they propagate in the same direction. However, in reality this wave typically propagates at an angle to stress, which will decrease its sensitivity. This thesis reports a numerical model used to investigate the transducers parameters that can influence the directivity of the LCR wave and hence enable performance optimization when used for industrial applications. An orthogonal test method is used to study the transducer parameters which influence the LCR wave beams and this method provides a design tool that can be used to study and optimize multiple parameter experiments and identify which parameter or parameters are of most significance. The example considered simulation of the acoustic field in a 2-D water-steel model is obtained using a Spatial Fourier Analysis method. The significance of the effects of incident angle, the aperture and the center frequency of the transducer were studied. Results show that the aperture, the center frequency and the incident angle are the most important factors in controlling the directivity of the resulting LCR wave fields.

Structural, optical and dielectric properties of pure and chromium (Cr) doped nickel oxide nanoparticles

NASA Astrophysics Data System (ADS)

Gupta, Jhalak; Ahmed, Arham S.

2018-05-01

The pure and Cr doped nickel oxide (NiO) nanoparticles have been synthesized by cost effective co-precipitation method having nickel nitrate as initial precursor. The synthesized samples were characterized by X-Ray diffraction (XRD), UV-Visible Spectroscopy(UV-Vis) and LCR meter for structural, optical and dielectric properties respectively. The crystallite size of pure nickel oxide nanoparticles characterized by XRD using Debye Scherer's formula was found to be 21.7nm and the same decreases on increasing Cr concentration whereas optical and dielectric properties were analyzed by UV-Vis and LCR meter respectively. The energy band gaps were determined by UV-Vis using Tauc relation.

Endangered fish threatened by Asian fish tapeworm

USGS Publications Warehouse

Cole, Rebecca A.

2004-01-01

The Asian fish tapeworm, an exotic parasite, has invaded the endangered humpback chub (Gila cypha) population from the Colorado and Little Colorado Rivers in Grand Canyon, Arizona. This parasite causes disease and death in carp in aquaculture settings and may retard growth in hatchery-reared roundtail chub (Gila robusta). Other consequences include destruction and dysfunction of the intestinal lining and adverse changes to certain blood parameters. Introduced into the U.S. in the 1970s with imported grass carp (Ctenopharyngodon idella), the Asian fish tapeworm (Bothriocephalus acheilognathi) was discovered in the Little Colorado River (LCR) by 1990. The LCR is the main tributary to the Colorado River in Grand Canyon and is an important spawning area for humpback chub.

LCR circuit: new simple methods for measuring the equivalent series resistance of a capacitor and inductance of a coil

NASA Astrophysics Data System (ADS)

Ivković, Saša S.; Marković, Marija Z.; Ivković, Dragica Ž.; Cvetanović, Nikola

2017-09-01

Equivalent series resistance (ESR) represents the measurement of total energy loss in a capacitor. In this paper a simple method for measuring the ESR of ceramic capacitors based on the analysis of the oscillations of an LCR circuit is proposed. It is shown that at frequencies under 3300 Hz, the ESR is directly proportional to the period of oscillations. Based on the determined dependence of the ESR on the period, a method is devised and tested for measuring coil inductance. All measurements were performed using the standard equipment found in student laboratories, which makes both methods very suitable for implementation at high school and university levels.

NF-E2 and GATA binding motifs are required for the formation of DNase I hypersensitive site 4 of the human beta-globin locus control region.

PubMed Central

Stamatoyannopoulos, J A; Goodwin, A; Joyce, T; Lowrey, C H

1995-01-01

The beta-like globin genes require the upstream locus control region (LCR) for proper expression. The active elements of the LCR coincide with strong erythroid-specific DNase I-hypersensitive sites (HSs). We have used 5' HS4 as a model to study the formation of these HSs. Previously, we identified a 101 bp element that is required for the formation of this HS. This element binds six proteins in vitro. We now report a mutational analysis of the HS4 HS-forming element (HSFE). This analysis indicates that binding sites for the hematopoietic transcription factors NF-E2 and GATA-1 are required for the formation of the characteristic chromatin structure of the HS following stable transfection into murine erythroleukemia cells. Similarly arranged NF-E2 and GATA binding sites are present in the other HSs of the human LCR, as well as in the homologous mouse and goat sequences and the chicken beta-globin enhancer. A combination of DNase I and micrococcal nuclease sensitivity assays indicates that the characteristic erythroid-specific hypersensitivity of HS4 to DNase I is the result of tissue-specific alterations in both nucleosome positioning and tertiary DNA structure. Images PMID:7828582

DNA is structured as a linear "jigsaw puzzle" in the genomes of Arabidopsis, rice, and budding yeast.

PubMed

Liu, Yun-Hua; Zhang, Meiping; Wu, Chengcang; Huang, James J; Zhang, Hong-Bin

2014-01-01

Knowledge of how a genome is structured and organized from its constituent elements is crucial to understanding its biology and evolution. Here, we report the genome structuring and organization pattern as revealed by systems analysis of the sequences of three model species, Arabidopsis, rice and yeast, at the whole-genome and chromosome levels. We found that all fundamental function elements (FFE) constituting the genomes, including genes (GEN), DNA transposable elements (DTE), retrotransposable elements (RTE), simple sequence repeats (SSR), and (or) low complexity repeats (LCR), are structured in a nonrandom and correlative manner, thus leading to a hypothesis that the DNA of the species is structured as a linear "jigsaw puzzle". Furthermore, we showed that different FFE differ in their importance in the formation and evolution of the DNA jigsaw puzzle structure between species. DTE and RTE play more important roles than GEN, LCR, and SSR in Arabidopsis, whereas GEN and RTE play more important roles than LCR, SSR, and DTE in rice. The genes having multiple recognized functions play more important roles than those having single functions. These results provide useful knowledge necessary for better understanding genome biology and evolution of the species and for effective molecular breeding of rice.

Protection against bubonic and pneumonic plague with a single dose microencapsulated sub-unit vaccine.

PubMed

Elvin, Stephen J; Eyles, James E; Howard, Kenneth A; Ravichandran, Easwaran; Somavarappu, Satyanarayan; Alpar, H Oya; Williamson, E Diane

2006-05-15

Protection against virulent plague challenge by the parenteral and aerosol routes was afforded by a single administration of microencapsulated Caf1 and LcrV antigens from Yersinia pestis in BALB/c mice. Recombinant Caf1 and LcrV were individually encapsulated in polymeric microspheres, to the surface of which additional antigen was adsorbed. The microspheres containing either Caf1 or LcrV were blended and used to immunise mice on a single occasion, by either the intra-nasal or intra-muscular route. Both routes of immunisation induced systemic and local immune responses, with high levels of serum IgG being developed in response to both vaccine antigens. In Elispot assays, secretion of cytokines by spleen and draining lymph node cells was demonstrated, revealing activation of both Th1 and Th2 associated cytokines; and spleen cells from animals immunised by either route were found to proliferate in vitro in response to both vaccine antigens. Virulent challenge experiments demonstrated that non-invasive immunisation by intra-nasal instillation can provide strong systemic and local immune responses and protect against high level challenge. Microencapsulation of these vaccine antigens has the added advantage that controlled release of the antigens occurs in vivo, so that protective immunity can be induced after only a single immunising dose.

A Boy with an LCR3/4-Flanked 10q22.3q23.2 Microdeletion and Uncommon Phenotypic Features

PubMed Central

Petrova, E.; Neuner, C.; Haaf, T.; Schmid, M.; Wirbelauer, J.; Jurkutat, A.; Wermke, K.; Nanda, I.; Kunstmann, E.

2014-01-01

The recurrent 10q22.3q23.2 deletion with breakpoints within low copy repeats 3 and 4 is a rare genomic disorder, reported in only 13 patients to date. The phenotype is rather uncharacteristic, which makes a clinical diagnosis difficult. A phenotypic feature described in almost all patients is a delay in speech development, albeit systematic studies are still pending. In this study, we report on a boy with an LCR3/4-flanked 10q22.3q23.2 deletion exhibiting an age-appropriate language development evaluated by a standardized test at an age of 2 years and 3 months. The boy was born with a cleft palate – a feature not present in any of the patients described before. Previously reported cases are reviewed, and the role of the BMPR1A gene is discussed. The phenotype of patients with an LCR3/4-flanked 10q22.3q23.2 deletion can be rather variable, so counseling the families regarding the prognosis of an affected child should be done with caution. Long-term studies of affected children are needed to delineate the natural history of this rare disorder. PMID:24550761

Validating Fricke dosimetry for the measurement of absorbed dose to water for HDR 192Ir brachytherapy: a comparison between primary standards of the LCR, Brazil, and the NRC, Canada.

PubMed

Salata, Camila; David, Mariano Gazineu; de Almeida, Carlos Eduardo; El Gamal, Islam; Cojocaru, Claudiu; Mainegra-Hing, Ernesto; McEwen, Malcom

2018-04-05

Two Fricke-based absorbed dose to water standards for HDR Ir-192 dosimetry, developed independently by the LCR in Brazil and the NRC in Canada have been compared. The agreement in the determination of the dose rate from a HDR Ir-192 source at 1 cm in a water phantom was found to be within the k  =  1 combined measurement uncertainties of the two standards: D NRC /D LCR   =  1.011, standard uncertainty  =  2.2%. The dose-based standards also agreed within the uncertainties with the manufacturer's stated dose rate value, which is traceable to a national standard of air kerma. A number of possible influence quantities were investigated, including the specific method for producing the ferrous-sulphate Fricke solution, the geometry of the holder, and the Monte Carlo code used to determine correction factors. The comparison highlighted the lack of data on the determination of G(Fe 3+ ) in this energy range and the possibilities for further development of the holders used to contain the Fricke solution. The comparison also confirmed the suitability of Fricke dosimetry for Ir-192 primary standard dose rate determinations at therapy dose levels.

Deficiency in the Heat Stress Response Could Underlie Susceptibility to Metabolic Disease

PubMed Central

Rogers, Robert S.; Morris, E. Matthew; Wheatley, Joshua L.; Archer, Ashley E.; McCoin, Colin S.; White, Kathleen S.; Wilson, David R.; Meers, Grace M.E.; Koch, Lauren G.; Britton, Steven L.

2016-01-01

Heat treatment (HT) effectively prevents insulin resistance and glucose intolerance in rats fed a high-fat diet (HFD). The positive metabolic actions of heat shock protein 72 (HSP72), which include increased oxidative capacity and enhanced mitochondrial function, underlie the protective effects of HT. The purpose of this study was to test the ability of HSP72 induction to mitigate the effects of consumption of a short-term 3-day HFD in rats selectively bred to be low-capacity runners (LCRs) and high-capacity runners (HCRs)—selective breeding that results in disparate differences in intrinsic aerobic capacity. HCR and LCR rats were fed a chow or HFD for 3 days and received a single in vivo HT (41°C, for 20 min) or sham treatment (ST). Blood, skeletal muscles, liver, and adipose tissues were harvested 24 h after HT/ST. HT decreased blood glucose levels, adipocyte size, and triglyceride accumulation in liver and muscle and restored insulin sensitivity in glycolytic muscles from LCR rats. As expected, HCR rats were protected from the HFD. Importantly, HSP72 induction was decreased in LCR rats after only 3 days of eating the HFD. Deficiency in the highly conserved stress response mediated by HSPs could underlie susceptibility to metabolic disease with low aerobic capacity. PMID:27554472

Degree of conversion of two lingual retainer adhesives cured with different light sources.

PubMed

Usümez, Serdar; Büyükyilmaz, Tamer; Karaman, Ali Ihya; Gündüz, Beniz

2005-04-01

The aim of this study was to evaluate the degree of conversion (DC) of two lingual retainer adhesives, Transbond Lingual Retainer (TLR) and Light Cure Retainer (LCR), cured with a fast halogen light, a plasma arc light and a light-emitting diode (LED) at various curing times. A conventional halogen light served as the control. One hundred adhesive samples (five per group) were cured for 5, 10 or 15 seconds with an Optilux 501 (fast halogen light), for 3, 6 or 9 seconds with a Power Pac (plasma arc light), or for 10, 20 or 40 seconds with an Elipar Freelight (LED). Samples cured for 40 seconds with the conventional halogen lamp were used as the controls. Absorbance peaks were recorded using Fourier transform infrared (FT-IR) spectroscopy. DC values were calculated. Data were analysed using Kruskal-Wallis and Mann-Whitney U-tests. For the TLR, the highest DC values were achieved in 6 and 9 seconds with the plasma arc light. Curing with the fast halogen light for 15 seconds and with the LED for 40 seconds produced statistically similar DC values, but these were lower than those with the plasma arc light. All of these light exposures yielded a statistically significantly higher DC than 40 seconds of conventional halogen light curing. The highest DC value for the LCR was achieved in 15 seconds with the fast halogen light, then the plasma arc light curing for 6 seconds. These two combinations produced a statistically significantly higher DC when compared with the 40 seconds of conventional halogen light curing. The lowest DC for the LCR was achieved with 10 seconds of LED curing. The overall DC of the LCR was significantly higher than that of the TLR. The results suggest that a similar or higher DC than the control values could be achieved in 6-9 seconds by plasma arc curing, in 10-15 seconds by fast halogen curing or in 20 seconds by LED curing.

Advantages of laparoscopic compared to conventional surgery are not related to an innate immune response of peritoneal immune activation: an animal study in rats.

PubMed

Lingohr, Philipp; Dohmen, Jonas; Matthaei, Hanno; Schwandt, Timo; Stein, Kathy; Hong, Gun-Soo; Steitz, Julia; Longerich, Thomas; Bölke, Edwin; Wehner, Sven; Kalff, Jörg C

2017-06-01

Laparoscopic surgery (LS) has proved superior compared to conventional surgery (CS) regarding morbidity, length of hospital stay, rate of wound infection and time until recovery. An improved preservation of the postoperative immune function is assumed to contribute to these benefits though the role of the local peritoneal immune response is still poorly understood. Our study investigates the peritoneal immune response subsequent to abdominal surgery and compares it between laparoscopic and conventional surgery to find an immunological explanation for the clinically proven benefits of LS. Wistar rats (N = 140) underwent laparoscopic cecum resection (LCR; N = 28), conventional cecum resection (CCR; N = 28), laparoscopic sham operation (LSO; N = 28), conventional sham operation (CSO; N = 28), or no surgical treatment (CTRL; N = 28). Postoperatively, peritoneal lavages were performed, leukocytes isolated and analyzed regarding immune function and phagocytosis activity. Immune function was inhibited postoperatively in animals undergoing LCR or CCR compared to CTRL reflected by a lower TNF-α (CTRL 3956.65 pg/ml, LCR 2018.48 pg/ml (p = 0.023), CCR 2793.78 pg/ml (n.s.)) and IL-6 secretion (CTRL 625.84 pg/ml, LCR 142.84 pg/ml (p = 0.009), CCR 169.53 pg/ml (p = 0.01)). Phagocytosis was not affected in rats undergoing any kind of surgery compared to CTRL. Neither cytokine secretion nor phagocytosis activity differed significantly between laparoscopic and conventional surgery. According to our findings the benefits associated with LS compared to CS cannot be explained by differences in the postoperative peritoneal innate immune response. Further studies are needed to elucidate the causes for a more favorable postoperative outcome in patients after LS compared to CS.

Real-Time Characterization of Virulence Factor Expression in Yersinia pestis Using a Green Fluorescent Protein Reporter System

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

Forde, C; Rocco, J; Fitch, J P

2004-06-09

A real-time reporter system was developed to monitor the thermal induction of virulence factors in Yersinia pestis. The reporter system consists of a plasmid in Y. pestis in which the expression of green fluorescent protein (GFP) is under the control of the promoters for six virulence factors, yopE, sycE, yopK, yopT, yscN, and lcrE/yopN, which are all components of the Type III secretion virulence mechanism of Y. pestis. Induction of the expression of these genes in vivo was determined by the increase in fluorescence intensity of GFP in real time. Basal expression levels observed for the Y. pestis promoters, expressedmore » as percentages of the positive control with GFP under the control of the lac promoter, were: yopE (15%), sycE (15%), yopK (13%), yopT (4%), lcrE (3.3%) and yscN (0.8%). The yopE reporter showed the strongest gene induction following temperature transition from 26 C to 37 C. The induction levels of the other virulence factors, expressed as percentages of yopE induction, were: yopK (57%), sycE (9%), yscN (3%), lcrE (3%), and yopT (2%). The thermal induction of each of these promoter fusions was repressed by calcium, and the ratios of the initial rates of thermal induction without calcium supplementation compared to the rate with calcium supplementation were: yopE (11 fold), yscN (7 fold), yopK (6 fold), lcrE (3 fold), yopT (2 fold), and sycE (2 fold). This work demonstrates a novel approach to quantify gene induction and provides a method to rapidly determine the effects of external stimuli on expression of Y. pestis virulence factors in real time, in living cells.« less

Fine-tuning synthesis of Yersinia pestis LcrV from runaway-like replication balanced-lethal plasmid in a Salmonella enterica serovar typhimurium vaccine induces protection against a lethal Y. pestis challenge in mice.

PubMed

Torres-Escobar, Ascención; Juárez-Rodríguez, María Dolores; Gunn, Bronwyn M; Branger, Christine G; Tinge, Steven A; Curtiss, Roy

2010-06-01

A balanced-lethal plasmid expression system that switches from low-copy-number to runaway-like high-copy-number replication (pYA4534) was constructed for the regulated delayed in vivo synthesis of heterologous antigens by vaccine strains. This is an antibiotic resistance-free maintenance system containing the asdA gene (essential for peptidoglycan synthesis) as a selectable marker to complement the lethal chromosomal DeltaasdA allele in live recombinant attenuated Salmonella vaccines (RASVs) such as Salmonella enterica serovar Typhimurium strain chi9447. pYA4534 harbors two origins of replication, pSC101 and pUC (low and high copy numbers, respectively). The pUC replication origin is controlled by a genetic switch formed by the operator/promoter of the P22 cro gene (O/P(cro)) (P(R)), which is negatively regulated by an arabinose-inducible P22 c2 gene located on both the plasmid and the chromosome (araC P(BAD) c2). The absence of arabinose, which is unavailable in vivo, triggers replication to a high-copy-number plasmid state. To validate these vector attributes, the Yersinia pestis virulence antigen LcrV was used to develop a vaccine against plague. An lcrV sequence encoding amino acids 131 to 326 (LcrV196) was optimized for expression in Salmonella, flanked with nucleotide sequences encoding the signal peptide (SS) and the carboxy-terminal domain (CT) of beta-lactamase, and cloned into pYA4534 under the control of the P(trc) promoter to generate plasmid pYA4535. Our results indicate that the live Salmonella vaccine strain chi9447 harboring pYA4535 efficiently stimulated a mixed Th1/Th2 immune response that protected mice against lethal challenge with Y. pestis strain CO92 introduced through either the intranasal or subcutaneous route.

Fine-Tuning Synthesis of Yersinia pestis LcrV from Runaway-Like Replication Balanced-Lethal Plasmid in a Salmonella enterica Serovar Typhimurium Vaccine Induces Protection against a Lethal Y. pestis Challenge in Mice▿

PubMed Central

Torres-Escobar, Ascención; Juárez-Rodríguez, María Dolores; Gunn, Bronwyn M.; Branger, Christine G.; Tinge, Steven A.; Curtiss, Roy

2010-01-01

A balanced-lethal plasmid expression system that switches from low-copy-number to runaway-like high-copy-number replication (pYA4534) was constructed for the regulated delayed in vivo synthesis of heterologous antigens by vaccine strains. This is an antibiotic resistance-free maintenance system containing the asdA gene (essential for peptidoglycan synthesis) as a selectable marker to complement the lethal chromosomal ΔasdA allele in live recombinant attenuated Salmonella vaccines (RASVs) such as Salmonella enterica serovar Typhimurium strain χ9447. pYA4534 harbors two origins of replication, pSC101 and pUC (low and high copy numbers, respectively). The pUC replication origin is controlled by a genetic switch formed by the operator/promoter of the P22 cro gene (O/Pcro) (PR), which is negatively regulated by an arabinose-inducible P22 c2 gene located on both the plasmid and the chromosome (araC PBAD c2). The absence of arabinose, which is unavailable in vivo, triggers replication to a high-copy-number plasmid state. To validate these vector attributes, the Yersinia pestis virulence antigen LcrV was used to develop a vaccine against plague. An lcrV sequence encoding amino acids 131 to 326 (LcrV196) was optimized for expression in Salmonella, flanked with nucleotide sequences encoding the signal peptide (SS) and the carboxy-terminal domain (CT) of β-lactamase, and cloned into pYA4534 under the control of the Ptrc promoter to generate plasmid pYA4535. Our results indicate that the live Salmonella vaccine strain χ9447 harboring pYA4535 efficiently stimulated a mixed Th1/Th2 immune response that protected mice against lethal challenge with Y. pestis strain CO92 introduced through either the intranasal or subcutaneous route. PMID:20308296

Distribution of four viruses in single and mixed infections within infected watermelon plants in Florida.

PubMed

Turechek, William W; Kousik, Chandrasekar S; Adkins, Scott

2010-11-01

Whitefly-transmitted Squash vein yellowing virus (SqVYV) and Cucurbit leaf crumple virus (CuLCrV) and aphid-transmitted Papaya ringspot virus type W (PRSV-W) have had serious impact on watermelon production in southwest and west-central Florida in the past 5 years. Tissue-blot nucleic acid hybridization assays were developed for simple, high-throughput detection of these three viruses as well as Cucurbit yellow stunting disorder virus (CYSDV), which was first reported in Florida in 2008. To determine virus distribution within plants, we collected 80 entire plants just before or during the harvest period in a systematic sample, 20 each on 11 April, 18 April, 26 April, and 3 May 2007, from a fruiting commercial watermelon field near Immokalee, FL showing symptoms of infection by SqVYV, CuLCrV, and PRSV-W and, possibly, CYSDV. This was followed by a sampling of five plants collected at harvest showing symptoms of virus infection on 11 October 2007 in a different commercial planting located in Duette, FL. Tissue prints were made from cross sections of watermelon plants from the crowns through the tips at 0.6-m intervals on nylon membranes and nucleic acid hybridization assays were used for virus detection. Results from testing crown tissue showed that SqVYV, CuLCrV, and PRSV-W were present in ≈37, 44, and 54%, respectively, of the 80 plants collected over the four sampling dates from the first field. For individual vines diagnosed with SqVYV, the distribution of SqVYV in vine tissue decreased proportionately with distance from the crown. The probability of detecting SqVYV was 70% at the base of the vine compared with 23% at the tip of the vine. In contrast, CuLCrV tended to be more evenly distributed throughout the plant, with ≈10% higher probability of detection at the growing tip relative to the crown of the plant. The distribution of PRSV-W resembled that of SqVYV but with ≈20% higher probability of detection at the tip of the vine. Similar trends were detected in the smaller sampling; however, CYSDV was also detected in three of the plants. Overall, the results indicated that SqVYV and PRSV-W were distributed differently than CuLCrV in watermelon plants, and this difference has implications on how samples should be collected and may affect vector acquisition and transmission of these viruses.

The critical distance in laser-induced plasmas: an operative definition

NASA Astrophysics Data System (ADS)

Delle Side, D.; Giuffreda, E.; Nassisi, V.

2016-05-01

We propose a method to estimate a precise value for the critical distance Lcr after which three-body recombination stops to produce charge losses in an expanding laser-induced plasma. We show in particular that the total charge collected has a ``reversed sigmoid'' shape as a function of the target-to-detector distance. Fitting the total charge data with a logistic related function, we could consider as Lcr the intercept of the tangent to this curve in its inflection point. Furthermore, this value scales well with theoretical predictions. From the application point of view, this could be of great practical interest, since it provide a reliable way to precisely determine the geometry of the extraction system in Laser Ion Sources.

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

Vargas, Asticio; Center for Optics and Photonics, Universidad de Concepción, Casilla 4016, Concepción; Mar Sánchez-López, María del

Multiple-beam Fabry-Perot (FP) interferences occur in liquid crystal retarders (LCR) devoid of an antireflective coating. In this work, a highly accurate method to obtain the spectral retardance of such devices is presented. On the basis of a simple model of the LCR that includes FP effects and by using a voltage transfer function, we show how the FP features in the transmission spectrum can be used to accurately retrieve the ordinary and extraordinary spectral phase delays, and the voltage dependence of the latter. As a consequence, the modulation characteristics of the device are fully determined with high accuracy by meansmore » of a few off-state physical parameters which are wavelength-dependent, and a single voltage transfer function that is valid within the spectral range of characterization.« less

Experiments and Demonstrations in Physics: Bar-Ilan Physics Laboratory (2nd Edition)

NASA Astrophysics Data System (ADS)

Kraftmakher, Yaakov

2014-08-01

The following sections are included: * Data-acquisition systems from PASCO * ScienceWorkshop 750 Interface and DataStudio software * 850 Universal Interface and Capstone software * Mass on spring * Torsional pendulum * Hooke's law * Characteristics of DC source * Digital storage oscilloscope * Charging and discharging a capacitor * Charge and energy stored in a capacitor * Speed of sound in air * Lissajous patterns * I-V characteristics * Light bulb * Short time intervals * Temperature measurements * Oersted's great discovery * Magnetic field measurements * Magnetic force * Magnetic braking * Curie's point I * Electric power in AC circuits * Faraday's law of induction I * Self-inductance and mutual inductance * Electromagnetic screening * LCR circuit I * Coupled LCR circuits * Probability functions * Photometric laws * Kirchhoff's rule for thermal radiation * Malus' law * Infrared radiation * Irradiance and illuminance

Pneumonic plague pathogenesis and immunity in Brown Norway rats.

PubMed

Anderson, Deborah M; Ciletti, Nancy A; Lee-Lewis, Hanni; Elli, Derek; Segal, Joshua; DeBord, Kristin L; Overheim, Katie A; Tretiakova, Maria; Brubaker, Robert R; Schneewind, Olaf

2009-03-01

The Brown Norway rat was recently described as a bubonic plague model that closely mimics human disease. We therefore evaluated the Brown Norway rat as an alternative small animal model for pneumonic plague and characterized both the efficacy and potency of vaccine candidates. When infected by intranasal instillation, these rats rapidly developed fatal pneumonic plague within 2 to 4 days of infection. Plague disease was characterized by severe alveolar edema and vascular hemorrhage in the lung in addition to fulminant necrotizing pneumonia caused by massive bacterial replication and inflammation. Twenty-four hours before death, animals developed systemic disease with an apparent delayed inflammatory response. We evaluated the ability of the protective antigen, LcrV, and a mutant derivative, V10, to protect these rats from pneumonic plague. Both were highly effective vaccines because complete protection was observed at challenge doses of 7500 LD(50). Antibody analyses suggested stronger potency of V10 immune sera compared with LcrV in the passive transfer of immunity to bubonic plague, with multiple neutralizing epitopes in LcrV. Taken together, these data demonstrate the effectiveness of inhibiting type III secretion in the prevention of pneumonic plague in rats and reveal critical contributions from both the cellular and humoral immune systems. Thus, the Brown Norway rat is an appealing alternative small animal model for the study of pneumonic plague pathogenesis and immunity.

Prevention of pneumonic plague in mice, rats, guinea pigs and non-human primates with clinical grade rV10, rV10-2 or F1-V vaccines

PubMed Central

Quenee, Lauriane E.; Ciletti, Nancy A.; Elli, Derek; Hermanas, Timothy M.; Schneewind, Olaf

2012-01-01

Yersinia pestis causes plague, a disease with high mortality in humans that can be transmitted by fleabite or aerosol. A US Food and Drug Administration (FDA)-licensed plague vaccine is currently not available. Vaccine developers have focused on two subunits of Y. pestis: LcrV, a protein at the tip of type III secretion needles, and F1, the fraction 1 pilus antigen. F1-V, a hybrid generated via translational fusion of both antigens, is being developed for licensure as a plague vaccine. The rV10 vaccine is a non-toxigenic variant of LcrV lacking residues 271–300. Here we developed Current Good Manufacturing Practice (cGMP) protocols for rV10. Comparison of clinical grade rV10 with F1-V did not reveal significant differences in plague protection in mice, guinea pigs or cynomolgus macaques. We also developed cGMP protocols for rV10-2, a variant of rV10 with an altered affinity tag. Immunization with rV10-2 adsorbed to aluminum hydroxide elicited antibodies against LcrV and conferred pneumonic plague protection in mice, rats, guinea pigs, cynomolgus macaques and African Green monkeys. The data support further development of rV10-2 for FDA Investigational New Drug (IND) authorization review and clinical testing. PMID:21763383

Clinical relevance of KRAS mutation detection in metastatic colorectal cancer treated by Cetuximab plus chemotherapy

PubMed Central

Di Fiore, F; Blanchard, F; Charbonnier, F; Le Pessot, F; Lamy, A; Galais, M P; Bastit, L; Killian, A; Sesboüé, R; Tuech, J J; Queuniet, A M; Paillot, B; Sabourin, J C; Michot, F; Michel, P; Frebourg, T

2007-01-01

The predictive value of KRAS mutation in metastatic colorectal cancer (MCRC) patients treated with cetuximab plus chemotherapy has recently been suggested. In our study, 59 patients with a chemotherapy-refractory MCRC treated with cetuximab plus chemotherapy were included and clinical response was evaluated according to response evaluation criteria in solid tumours (RECIST). Tumours were screened for KRAS mutations using first direct sequencing, then two sensitive methods based on SNaPshot and PCR-ligase chain reaction (LCR) assays. Clinical response was evaluated according to gene mutations using the Fisher exact test. Times to progression (TTP) were calculated using the Kaplan–Meier method and compared with log-rank test. A KRAS mutation was detected in 22 out of 59 tumours and, in six cases, was missed by sequencing analysis but detected using the SNaPshot and PCR-LCR assays. Remarkably, no KRAS mutation was found in the 12 patients with clinical response. KRAS mutation was associated with disease progression (P=0.0005) and TTP was significantly decreased in mutated KRAS patients (3 vs 5.5 months, P=0.015). Our study confirms that KRAS mutation is highly predictive of a non-response to cetuximab plus chemotherapy in MCRC and highlights the need to use sensitive molecular methods, such as SNaPshot or PCR-LCR assays, to ensure an efficient mutation detection. PMID:17375050

Investigating the compatibility of PEEK polymer for the fabrication of sample cells for use in muon spin spectroscopy

NASA Astrophysics Data System (ADS)

Chandrasena, L.; McKenzie, I.; Brodovitch, J.-C.; Mozafari, M.; Cottrell, S. P.; Percival, P. W.

2014-12-01

Polyether ether ketone (PEEK) is a thermoplastic polymer with a wide range of applications due to its chemical inertness and thermal stability, and for these reasons sample cells for gas and liquid phase μSR have been constructed from PEEK. Muon levelcrossing resonance (μLCR) studies of PEEK revealed a broad, strong μLCR signal that, we hypothesize, is due to multiple overlapping resonances from the various muonium (Mu) adducts of PEEK. To investigate this, two monomer units from PEEK (4,4'-dihydroxybenzophenone and para-dimethoxybenzene) were studied in solution using transverse-field muon spin rotation (TF-μSR) and μLCR. Two different muoniated radicals were formed by Mu addition to 4,4/- dihydroxybenzophenone and one radical was formed in para-dimethoxybenzene. The μSR spectra were assigned by comparing the experimentally measured muon and proton hyperfine coupling constants with values calculated for the possible structures using Gaussian-09 software with the B3LYP functional and 6-31G basis set. Good agreement was found for cyclohexadienyl- type radicals formed by Mu addition to the benzene rings of the monomer units. We can also infer that these radicals are being formed in PEEK, and based on this we conclude that sample cells made of PEEK are unsuitable for many types of μSR experiment.

Pro-inflammatory and oxidative stress pathways which compromise sperm motility and survival may be altered by L-carnitine

PubMed Central

Helal, Gouda K; Al-Yahya, Abdulaziz A; Aleisa, Abdulaziz M; Al-Rejaie, Salim S; Al-Bakheet, Saleh A

2009-01-01

The testis is an immunologically privileged organ. Sertoli cells can form a blood-testis barrier and protect sperm cells from self-immune system attacks. Spermatogenesis may be inhibited by severe illness, bacterial infections and chronic inflammatory diseases but the mechanism(s) is poorly understood. Our objective is to help in understanding such mechanism(s) to develop protective agents against temporary or permanent testicular dysfunction. Lipopolysaccaride (LPS) is used as a model of animal sepsis while L-carnitine (LCR) is used as a protective agent. A total of 60 male Swiss albino rats were divided into four groups (15/group). The control group received Saline; the 2nd group was given LCR (500 mg/kg i.p, once). The third group was treated with LPS (5 mg/kg i.p once) and the fourth group received LCR then LPS after three hours. From each group, five rats were used for histopathological examination. Biochemical parameters were assessed in the remaining ten rats. At the end of the experiment, animals were lightly anaesthetized with ether where blood samples were collected and testes were dissected on ice. Sperm count and motility were evaluated from cauda epididymis in each animal. Also, oxidative stress was evaluated by measuring testicular contents of reduced glutathione (GSH), malondialdehyde (MDA) and 8-hydroxydeoxyguanosine (8-HDG, the DNA adduct for oxidative damage) in testicular DNA. The pro-inflammatory mediator nitric oxide (NO) in addition to lactate dehydrogenase (LDHx) isoenzyme-x activity as an indicator for normal spermatozoal metabolism were assessed in testicular homogenate. Serum interlukin (IL)-2 level was also assessed as a marker for T-helper cell function. The obtained data revealed that LPS induced marked reductions in sperm's count and motility, obstruction in seminiferous tubules, hypospermia and dilated congested blood vessels in testicular sections concomitant with decreased testicular GSH content and LDHx activity. Moreover, the testicular levels of MDA, 8-HDG (in testicular DNA) and NO as well as serum IL-2 level were increased. Administration of LCR before LPS returned both sperm count and motility to normal levels. Also, contents of testicular GSH, MDA, 8-HDG and NO returned back to the corresponding control values. In addition, serum IL-2 level as well as histological abnormalities were markedly improved in LCR + LPS-treated rats. In conclusion, LPS increased proinflammatory and oxidative stress markers in the testis leading to a marked testicular dysfunction. L-carnitine administration ameliorates these effects by antioxidant and/or anti-inflammatory mechanisms suggesting a protective role against male infertility in severely infected or septic patients. PMID:20357928

Optimal Corrosion Control Treatment Evaluation Technical Recommendations

EPA Pesticide Factsheets

This document provides technical recommendations that both systems and primacy agencies can use to comply with LCR CCT requirements and effective evaluation and designation of optimal corrosion control treatment (OCCT).

The mid-Holocene to present large-scale morphodynamic and coupled fluvial-tidal sedimentologic evolution of the Lower Columbia River, WA/OR, USA

NASA Astrophysics Data System (ADS)

Prokocki, E.; Best, J.; Ashworth, P. J.; Parsons, D. R.; Sambrook Smith, G.; Nicholas, A. P.; Simpson, C.; Wang, H.; Sandbach, S.; Keevil, C.

2015-12-01

Optically stimulated luminescence (OSL) dating of four deep sediment cores (≤ 20m depth), in conjunction with shallow vibracores (≤ 6m depth), obtained from mid-channel bars in the lower Columbia River (LCR), USA, provides new insights into the mid-Holocene to present geomorphic and coupled sedimentological evolution of the LCR fluvial-tidal zone. These data reveal that the relatively coarse-grained basal sediments of mid-channel bars positioned across the LCR tidal-fluvial hydraulic regime were deposited at c. 2.5 to 2.0 ka, and not at c. 8.0 ka as previously reported. Thus, these younger depositional ages of basal sediments relative to previous studies coupled with the overall sedimentary architecture of these bars, and the absence of a temporal lag in the timing of basal sedimentation between bars located from river kilometer 51.1 to 29.3, challenges existing models that these bars represent: (a) estuarine tidal-bars, or (b) bay-head deltaic deposits. Within the context of post glacial Holocene sea-level rise, our results suggest these bars represent vertical construction of a LCR fluvial top-set from c. 2.5- 2.0 ka to the present, as the regional rate of sea-level rise slowed to ≤ 1.4 mmyr-1. Within this geomorphic context, two tidal-fluvial sedimentological signatures can be identified: (i) in the downstream direction, basal bar deposits incorporate a larger percentage of finer-grained interbeds, and (ii) vertically stacked silt/very-fine sand draped current ripple cross-laminae become prevalent from approximately 5 m in depth to the bar surfaces. The preservation of finer-grained interbeds within basal bar deposits is reasoned to be caused by the flocculation and settling of suspended sediment enhanced by the turbidity maximum. The stacked draped current ripple cross-laminae are interpreted to result from tidal-currents generating asymmetric current ripples that were draped by fine-sediment entrained by wind-waves, which fell-out of suspension during reduced wave activity, slackwater intervals, and periods when the turbidity maximum was active.

Beat-to-Beat Variation in Periodicity of Local Calcium Releases Contributes to Intrinsic Variations of Spontaneous Cycle Length in Isolated Single Sinoatrial Node Cells

PubMed Central

Monfredi, Oliver; Maltseva, Larissa A.; Spurgeon, Harold A.; Boyett, Mark R.; Lakatta, Edward G.; Maltsev, Victor A.

2013-01-01

Spontaneous, submembrane local Ca2+ releases (LCRs) generated by the sarcoplasmic reticulum in sinoatrial nodal cells, the cells of the primary cardiac pacemaker, activate inward Na+/Ca2+-exchange current to accelerate the diastolic depolarization rate, and therefore to impact on cycle length. Since LCRs are generated by Ca2+ release channel (i.e. ryanodine receptor) openings, they exhibit a degree of stochastic behavior, manifested as notable cycle-to-cycle variations in the time of their occurrence. Aim The present study tested whether variation in LCR periodicity contributes to intrinsic (beat-to-beat) cycle length variability in single sinoatrial nodal cells. Methods We imaged single rabbit sinoatrial nodal cells using a 2D-camera to capture LCRs over the entire cell, and, in selected cells, simultaneously measured action potentials by perforated patch clamp. Results LCRs begin to occur on the descending part of the action potential-induced whole-cell Ca2+ transient, at about the time of the maximum diastolic potential. Shortly after the maximum diastolic potential (mean 54±7.7 ms, n = 14), the ensemble of waxing LCR activity converts the decay of the global Ca2+ transient into a rise, resulting in a late, whole-cell diastolic Ca2+ elevation, accompanied by a notable acceleration in diastolic depolarization rate. On average, cells (n = 9) generate 13.2±3.7 LCRs per cycle (mean±SEM), varying in size (7.1±4.2 µm) and duration (44.2±27.1 ms), with both size and duration being greater for later-occurring LCRs. While the timing of each LCR occurrence also varies, the LCR period (i.e. the time from the preceding Ca2+ transient peak to an LCR’s subsequent occurrence) averaged for all LCRs in a given cycle closely predicts the time of occurrence of the next action potential, i.e. the cycle length. Conclusion Intrinsic cycle length variability in single sinoatrial nodal cells is linked to beat-to-beat variations in the average period of individual LCRs each cycle. PMID:23826247

Spontaneous, local diastolic subsarcolemmal calcium releases in single, isolated guinea-pig sinoatrial nodal cells.

PubMed

Sirenko, Syevda G; Yang, Dongmei; Maltseva, Larissa A; Kim, Mary S; Lakatta, Edward G; Maltsev, Victor A

2017-01-01

Uptake and release calcium from the sarcoplasmic reticulum (SR) (dubbed "calcium clock"), in the form of spontaneous, rhythmic, local diastolic calcium releases (LCRs), together with voltage-sensitive ion channels (membrane clock) form a coupled system that regulates the action potential (AP) firing rate. LCRs activate Sodium/Calcium exchanger (NCX) that accelerates diastolic depolarization and thus participating in regulation of the time at which the next AP will occur. Previous studies in rabbit SA node cells (SANC) demonstrated that the basal AP cycle length (APCL) is tightly coupled to the basal LCR period (time from the prior AP-induced Ca2+ transient to the diastolic LCR occurrence), and that this coupling is further modulated by autonomic receptor stimulation. Although spontaneous LCRs during diastolic depolarization have been reported in SANC of various species (rabbit, cat, mouse, toad), prior studies have failed to detect LCRs in spontaneously beating SANC of guinea-pig, a species that has been traditionally used in studies of cardiac pacemaker cell function. We performed a detailed investigation of whether guinea-pig SANC generate LCRs and whether they play a similar key role in regulation of the AP firing rate. We used two different approaches, 2D high-speed camera and classical line-scan confocal imaging. Positioning the scan-line beneath sarcolemma, parallel to the long axis of the cell, we found that rhythmically beating guinea-pig SANC do, indeed, generate spontaneous, diastolic LCRs beneath the surface membrane. The average key LCR characteristics measured in confocal images in guinea-pig SANC were comparable to rabbit SANC, both in the basal state and in the presence of β-adrenergic receptor stimulation. Moreover, the relationship between the LCR period and APCL was subtended by the same linear function. Thus, LCRs in guinea-pig SANC contribute to the diastolic depolarization and APCL regulation. Our findings indicate that coupled-clock system regulation of APCL is a general, species-independent, mechanism of pacemaker cell normal automaticity. Lack of LCRs in prior studies is likely explained by technical issues, as individual LCRs are small stochastic events occurring mainly near the cell border.

Effects of Aerobic Capacity on Thrombin-Induced Hydrocephalus and White Matter Injury.

PubMed

Ni, Wei; Gao, Feng; Zheng, Mingzhe; Koch, Lauren G; Britton, Steven L; Keep, Richard F; Xi, Guohua; Hua, Ya

2016-01-01

We have previously shown that intracerebral hemorrhage-induced brain injury is less in rats bred for high aerobic capacity (high capacity runners; HCR) compared with those bred for low aerobic capacity (low capacity runners; LCRs). Thrombin, an essential component in the coagulation cascade, is produced after cerebral hemorrhage. Intraventricular injection of thrombin causes significant hydrocephalus and white matter damage. In the present study, we examined the effect of exercise capacity on thrombin-induced hydrocephalus and white matter damage. Mid-aged (13-month-old) female LCRs (n = 13) and HCRs (n = 12) rats were used in this study. Rats received an intraventricular injection of thrombin (3 U, 50 μl). All rats underwent magnetic resonance imaging (MRI) at 24 h and were then euthanized for brain histology and Western blot. The mortalities were 20 % in LCRs and 33 % in HCRs after thrombin injection (p > 0.05). No rats died after saline injection. Intraventricular thrombin injection resulted in hydrocephalus and periventricular white matter damage as determined on MRI. In LCR rats, thrombin induced significant ventricle enlargement (23.0 ± 2.3 vs12.8 ± 1.9 mm(3) in LCR saline group; p < 0.01) and white matter lesion (9.3 ± 7.6 vs 0.6 ± 0.5 mm(3) in LCR saline group, p < 0.05). In comparison, in HCR rats thrombin induced less ventricular enlargement (17.3 ± 3.9 vs 23.0 ± 2.3 mm(3) in LCRs, p < 0.01) and smaller white matter lesions (2.6 ± 1.2 mm(3) vs 9.3 ± 7.6 mm(3) in LCRs, p < 0.05). In LCR rats, there was also upregulation of heat shock protein-32, a stress marker, and microglial activation in the periventricular white matter. These changes were significantly reduced in HCR rats. Intraventricular injection of thrombin caused more white matter damage and hydrocephalus in rats with low aerobic capacity. A differential effect of thrombin may contribute to differences in the effects of cerebral hemorrhage with aerobic capacity.

Spontaneous, local diastolic subsarcolemmal calcium releases in single, isolated guinea-pig sinoatrial nodal cells

PubMed Central

Sirenko, Syevda G.; Yang, Dongmei; Maltseva, Larissa A.; Kim, Mary S.; Lakatta, Edward G.

2017-01-01

Uptake and release calcium from the sarcoplasmic reticulum (SR) (dubbed “calcium clock”), in the form of spontaneous, rhythmic, local diastolic calcium releases (LCRs), together with voltage-sensitive ion channels (membrane clock) form a coupled system that regulates the action potential (AP) firing rate. LCRs activate Sodium/Calcium exchanger (NCX) that accelerates diastolic depolarization and thus participating in regulation of the time at which the next AP will occur. Previous studies in rabbit SA node cells (SANC) demonstrated that the basal AP cycle length (APCL) is tightly coupled to the basal LCR period (time from the prior AP-induced Ca2+ transient to the diastolic LCR occurrence), and that this coupling is further modulated by autonomic receptor stimulation. Although spontaneous LCRs during diastolic depolarization have been reported in SANC of various species (rabbit, cat, mouse, toad), prior studies have failed to detect LCRs in spontaneously beating SANC of guinea-pig, a species that has been traditionally used in studies of cardiac pacemaker cell function. We performed a detailed investigation of whether guinea-pig SANC generate LCRs and whether they play a similar key role in regulation of the AP firing rate. We used two different approaches, 2D high-speed camera and classical line-scan confocal imaging. Positioning the scan-line beneath sarcolemma, parallel to the long axis of the cell, we found that rhythmically beating guinea-pig SANC do, indeed, generate spontaneous, diastolic LCRs beneath the surface membrane. The average key LCR characteristics measured in confocal images in guinea-pig SANC were comparable to rabbit SANC, both in the basal state and in the presence of β-adrenergic receptor stimulation. Moreover, the relationship between the LCR period and APCL was subtended by the same linear function. Thus, LCRs in guinea-pig SANC contribute to the diastolic depolarization and APCL regulation. Our findings indicate that coupled-clock system regulation of APCL is a general, species-independent, mechanism of pacemaker cell normal automaticity. Lack of LCRs in prior studies is likely explained by technical issues, as individual LCRs are small stochastic events occurring mainly near the cell border. PMID:28945810

Lead and Copper Rule Revisions White Paper

EPA Pesticide Factsheets

The Lead and Copper Rule (LCR) Revisions White Paper provides examples of regulatory options to improve the existing rule. The paper highlights key challenges, opportunities, and analytical issues presented by these options.

SRO on the LCR Trip

ERIC Educational Resources Information Center

Trohanis, Pascal

1971-01-01

A look at multi-image instructional Films, including a film on production steps for a medium which blends the simultaneity of large three-screened still pictorials, motion picture film clips, and audio sounds." (Author/AK)

EPA Lead and Copper Rule State Response Documents

EPA Pesticide Factsheets

EPA recently sent letters to Governors and state environment and public health commissioners outlining specific steps to enhance oversight of LCR implementation. Below are the state responses to these letters EPA has received.

Some aspects of clinical relevance in the maturation of respiratory control in infants.

PubMed

Thach, Bradley T

2008-06-01

Two reflex mechanisms important for survival are discussed. Brain stem and cardiovascular mechanisms that are responsible for recovery from severe hypoxia (autoresuscitation) are important for survival in acutely hypoxic infants and adults. Failure of this mechanism may be important in sudden infant death syndrome (SIDS), because brain stem-mediated hypoxic gasping is essential for successful autoresuscitation and because SIDS infants appear to attempt to autoresuscitate just before death. A major function of another mechanism is to protect the airway from fluid aspiration. The various components of the laryngeal chemoreflex (LCR) change during maturation. The LCR is an important cause of prolonged apneic spells in infants. Consequently, it also may have a role in causing SIDS. Maturational changes and/or inadequacy of this reflex may be responsible for pulmonary aspiration and infectious pneumonia in both children and adults.

Influence of Hydrogen Peroxide, Lactic Acid, and Surfactants from Vaginal Lactobacilli on the Antibiotic Sensitivity of Opportunistic Bacteria.

PubMed

Sgibnev, Andrey; Kremleva, Elena

2017-06-01

We studied as hydrogen peroxide, lactic acid, or surfactants from clinical isolates of vaginal lactobacilli and cell-free supernatants from probiotic strain LCR35 can influence on the sensitivity of opportunistic bacteria to antibiotics. We found that the most effective in increasing sensitivity to antibiotics were hydrogen peroxide and surfactants or their combination but no lactic acid. In some cases, the effect of the composition of hydrogen peroxide and surfactants was clearly higher than the sum of effects of these substances alone. With using of the supernatant of LCR35 was shown that the combination of surfactant and lactate has greater effect compared with surfactants alone. In concluding, metabolites of vaginal lactobacilli are suitable for the role of "antibiotic assistants" and it can help solve the problems the antibiotic resistance.

Changes in mRNA levels for brain-derived neurotrophic factor after wheel running in rats selectively bred for high- and low-aerobic capacity

PubMed Central

Groves-Chapman, Jessica L.; Murray, Patrick S.; Stevens, Kristin L.; Monroe, Derek; Koch, Lauren G.; Britton, Steven L.; Holmes, Philip V.

2012-01-01

We evaluated levels of exercise-induced brain-derived neurotrophic factor (BDNF) messenger RNA (mRNA) within the hippocampal formation in rats selectively bred for 1) high intrinsic (i.e., untrained) aerobic capacity (High Capacity Runners, HCR), 2) low intrinsic aerobic capacity (Low Capacity Runners, LCR), and 3) unselected Sprague-Dawley (SD) rats with or without free access to running wheels for three weeks. The specific aim of the study was to determine whether a dose-response relationship exists between cumulative running distance and levels of BDNF mRNA. No additional treatments or behavioral manipulations were used. HCR, LCR, and SD rats were grouped by strain and randomly assigned to sedentary or activity (voluntary access to activity wheel) conditions. Animals were killed after 21 days of exposure to the assigned conditions. Daily running distances (mean ± standard deviation meters/d) during week three were: HCR (4726 ± 3220), SD (2293 ± 3461), LCR (672 ± 323). Regardless of strain, levels of BDNF mRNA in CA1 were elevated in wheel runners compared to sedentary rats and this difference persisted after adjustment for age (p=0.040). BDNF mRNA was not affected by intrinsic aerobic capacity and was not related to total running distance. The results support that BDNF mRNA expression is increased by unlimited access to activity wheel running for 3 weeks but is not dependent upon accumulated running distance. PMID:22024546

A Single Base Difference between Pit-1 Binding Sites at the hGH Promoter and Locus Control Region Specifies Distinct Pit-1 Conformations and Functions

PubMed Central

Shewchuk, Brian M.; Ho, Yugong; Liebhaber, Stephen A.; Cooke, Nancy E.

2006-01-01

Activation of the human growth hormone (hGH-N) gene in pituitary somatotropes is mediated by a locus control region (LCR). This LCR is composed of DNase I-hypersensitive sites (HS) located −14.5 kb to −32 kb relative to the hGH-N promoter. HSI, at −14.5 kb, is the dominant determinant of hGH-N expression and is essential for establishment of a 32-kb domain of histone acetylation that encompasses the active hGH locus. This activity is conferred by three binding sites for the POU domain transcription factor Pit-1. These Pit-1 elements are sufficient to activate hGH-N expression in the mouse pituitary. In contrast, Pit-1 sites at the hGH-N promoter are consistently unable to mediate similar activity. In the present study, we demonstrate that the functional difference between the promoter-proximal and the HSI Pit-1 binding sites can be attributed in part to a single base difference. This base affects the conformation of the Pit-1/DNA complex, and reciprocal exchange of the divergent bases between the two sets of Pit-1 elements results in a partial reversal of their transgenic activities. These data support a model in which the Pit-1 binding sites in the hGH LCR allosterically program the bound Pit-1 complex for chromatin activating functions. PMID:16914737

Singlet oxygen explicit dosimetry to predict local tumor control for HPPH-mediated photodynamic therapy

NASA Astrophysics Data System (ADS)

Penjweini, Rozhin; Kim, Michele M.; Ong, Yi Hong; Zhu, Timothy C.

2017-02-01

This preclinical study examines four dosimetric quantities (light fluence, photosensitizer photobleaching ratio, PDT dose, and reacted singlet oxygen ([1O2]rx)) to predict local control rate (LCR) for 2-(1-Hexyloxyethyl)-2-devinyl pyropheophorbide (HPPH)-mediated photodynamic therapy (PDT). Mice bearing radiation-induced fibrosarcoma (RIF) tumors were treated with different in-air fluences (135, 250 and 350 J/cm2) and in-air fluence rates (50, 75 and 150 mW/cm2) at 0.25 mg/kg HPPH and a drug-light interval of 24 hours using a 1 cm diameter collimated laser beam at 665 nm wavelength. A macroscopic model was used to calculate ([1O2]rx)) based on in vivo explicit dosimetry of the initial tissue oxygenation, photosensitizer concentration, and tissue optical properties. PDT dose was defined as a temporal integral of drug concentration and fluence rate (φ) at a 3 mm tumor depth. Light fluence rate was calculated throughout the treatment volume based on Monte-Carlo simulation and measured tissue optical properties. The tumor volume of each mouse was tracked for 30 days after PDT and Kaplan-Meier analyses for LCR were performed based on a tumor volume <=100 mm3, for four dose metrics: fluence, HPPH photobleaching rate, PDT dose, and ([1O2]rx)). The results of this study showed that ([1O2]rx)) is the best dosimetric quantity that can predict tumor response and correlate with LCR.

Ovariectomy results in differential shifts in gut microbiota in low versus high aerobic capacity rats

PubMed Central

Cox-York, Kimberly A; Sheflin, Amy M; Foster, Michelle T; Gentile, Christopher L; Kahl, Amber; Koch, Lauren G; Britton, Steven L; Weir, Tiffany L

2015-01-01

The increased risk for cardiometabolic disease with the onset of menopause is widely studied and likely precipitated by the decline in endogenous estradiol (E2), yet the precise mechanisms are unknown. The gut microbiome is involved in estrogen metabolism and has been linked to metabolic disease, suggesting its potential involvement in the postmenopausal phenotype. Furthermore, menopause-associated risk factors, as well as gut ecology, are altered with exercise. Therefore, we studied microbial changes in an ovariectomized (OVX vs. Sham) rat model of high (HCR) and low (LCR) intrinsic aerobic capacity (n = 8–10/group) in relation to changes in body weight/composition, glucose tolerance, and liver triglycerides (TG). Nine weeks after OVX, HCR rats were moderately protected against regional adipose tissue gain and liver TG accumulation (P < 0.05 for both). Microbial diversity and number of the Bacteroidetes phylum were significantly increased in LCR with OVX, but unchanged in HCR OVX relative to Sham. Plasma short-chain fatty acids (SCFA), produced by bacteria in the gut and recognized as metabolic signaling molecules, were significantly greater in HCR Sham relative to LCR Sham rats (P = 0.05) and were decreased with OVX in both groups. These results suggest that increased aerobic capacity may be protective against menopause-associated cardiometabolic risk and that gut ecology, and production of signaling molecules such as SCFA, may contribute to the mediation. PMID:26265751

Human papillomavirus type 18 variant lineages in United States populations characterized by sequence analysis of LCR-E6, E2, and L1 regions.

PubMed

Arias-Pulido, Hugo; Peyton, Cheri L; Torrez-Martínez, Norah; Anderson, D Nelson; Wheeler, Cosette M

2005-07-20

While HPV 16 variant lineages have been well characterized, the knowledge about HPV 18 variants is limited. In this study, HPV 18 nucleotide variations in the E2 hinge region were characterized by sequence analysis in 47 control and 51 tumor specimens. Fifty of these specimens were randomly selected for sequencing of an LCR-E6 segment and 20 samples representative of LCR-E6 and E2 sequence variants were examined across the L1 region. A total of 2770 nucleotides per HPV 18 variant genome were considered in this study. HPV 18 variant nucleotides were linked among all gene segments analyzed and grouped into three main branches: Asian-American (AA), European (E), and African (Af). These three branches were equally distributed among controls and cases and when stratified by Hispanic and non-Hispanic ethnicities. Among invasive cervical cancer cases, no significant differences in the three HPV variant branches were observed among ethnic groups or when stratified by histopathology (squamous vs. adenocarcinoma). The Af branch showed the greatest nucleotide variability when compared to the HPV 18 reference sequence and was more closely related to HPV 45 than either AA or E branches. Our data also characterize nucleotide and amino acid variations in the L1 capsid gene among HPV 18 variants, which may be relevant to vaccine strategies and subsequent studies of naturally occurring HPV 18 variants. Several novel HPV 18 nucleotide variations were identified in this study.

Inhibition of G9a methyltransferase stimulates fetal hemoglobin production by facilitating LCR/γ-globin looping.

PubMed

Krivega, Ivan; Byrnes, Colleen; de Vasconcellos, Jaira F; Lee, Y Terry; Kaushal, Megha; Dean, Ann; Miller, Jeffery L

2015-07-30

Induction of fetal hemoglobin (HbF) production in adult erythrocytes can reduce the severity of sickle cell disease and β-thalassemia. Transcription of β-globin genes is regulated by the distant locus control region (LCR), which is brought into direct gene contact by the LDB1/GATA-1/TAL1/LMO2-containing complex. Inhibition of G9a H3K9 methyltransferase by the chemical compound UNC0638 activates fetal and represses adult β-globin gene expression in adult human hematopoietic precursor cells, but the underlying mechanisms are unclear. Here we studied UNC0638 effects on β-globin gene expression using ex vivo differentiation of CD34(+) erythroid progenitor cells from peripheral blood of healthy adult donors. UNC0638 inhibition of G9a caused dosed accumulation of HbF up to 30% of total hemoglobin in differentiated cells. Elevation of HbF was associated with significant activation of fetal γ-globin and repression of adult β-globin transcription. Changes in gene expression were associated with widespread loss of H3K9me2 in the locus and gain of LDB1 complex occupancy at the γ-globin promoters as well as de novo formation of LCR/γ-globin contacts. Our findings demonstrate that G9a establishes epigenetic conditions preventing activation of γ-globin genes during differentiation of adult erythroid progenitor cells. In this view, manipulation of G9a represents a promising epigenetic approach for treatment of β-hemoglobinopathies.

MafK/NF-E2 p18 is required for beta-globin genes activation by mediating the proximity of LCR and active beta-globin genes in MEL cell line.

PubMed

Du, Mei-Jun; Lv, Xiang; Hao, De-Long; Zhao, Guo-Wei; Wu, Xue-Song; Wu, Feng; Liu, De-Pei; Liang, Chih-Chuan

2008-01-01

Evidences indicate that locus control region (LCR) of beta-globin spatially closes to the downstream active gene promoter to mediate the transcriptional activation by looping. DNA binding proteins may play an important role in the looping formation. NF-E2 is one of the key transcription factors in beta-globin gene transcriptional activation. To shed light on whether NF-E2 is involved in this process, DS19MafKsiRNA cell pools were established by specifically knocked down the expression of MafK/NF-E2 p18, one subunit of NF-E2 heterodimer. In the above cell pools, it was observed that the occupancy efficiency of NF-E2 on beta-globin gene locus and the expression level of beta-globin genes were decreased. H3 acetylation, H3-K4 methylation and the deposition of RNA polymerase II, but not the recruitment of GATA-1, were also found reduced at the beta-globin gene cluster. Chromosome Conformation Capture (3C) assay showed that the cross-linking frequency between the main NF-E2 binding site HS2 and downstream structural genes was reduced compared to the normal cell. This result demonstrated that MafK/NF-E2 p18 recruitment was involved in the physical proximity of LCR and active beta-globin genes upon beta-globin gene transcriptional activation.

Analysis of human papillomavirus 16 E6, E7 genes and Long Control Region in cervical samples from Uruguayan women.

PubMed

Ramas, Viviana; Mirazo, Santiago; Bonilla, Sylvia; Ruchansky, Dora; Arbiza, Juan

2018-05-15

This study aims to investigate the HPV16 variant distribution by sequence analyses of E6, E7 oncogenes and the Long Control Region (LCR), from cervical cells collected from Uruguayan women, and to reconstruct the phylogenetic relationships among variants. Forty-seven HPV16 variants, obtained from women with HSIL, LSIL, ASCUS and NILM cytological classes were analyzed for LCR and 12 were further studied for E6 and E7. Detailed sequence comparison, genetic heterogeneity analyses and phylogenetic reconstruction were performed. A high variability was observed among LCR sequences, which were distributed in 18 different variants. E6 and E7 sequences exhibited novel non-synonymous substitutions. Uruguayan sequences mainly belonged to the European lineage, and only 5 sequences clustered in non-European branches; 3 of them in the Asian-American and North-American linage and 2 in an African branch. Additionally, 6 new variants from European and African clusters were identified. HPV16 isolates mainly belonged to the European lineage, though strains from African and Asian-American lineages were also identified. Herein is reported for the first time the distribution and molecular characterization of HPV16 variants from Uruguay, providing novel insights on the molecular epidemiology of this infectious disease in the South America. A high variability among HPV 16 isolates mainly belonged to European lineage, provides an extensive sequence dataset from a country with high burden of cervical cancer. Copyright © 2018 Elsevier B.V. All rights reserved.

Lead and Copper Rule Long-Term Revisions

EPA Pesticide Factsheets

The goal for the Lead and Copper Rule (LCR) Long-Term Revisions is to improve public health protection provided by the by making substantive changes based on topics that were identified in the 2004 National Review.

GBA manager: an online tool for querying low-complexity regions in proteins.

PubMed

Bandyopadhyay, Nirmalya; Kahveci, Tamer

2010-01-01

Abstract We developed GBA Manager, an online software that facilitates the Graph-Based Algorithm (GBA) we proposed in our earlier work. GBA identifies the low-complexity regions (LCR) of protein sequences. GBA exploits a similarity matrix, such as BLOSUM62, to compute the complexity of the subsequences of the input protein sequence. It uses a graph-based algorithm to accurately compute the regions that have low complexities. GBA Manager is a user friendly web-service that enables online querying of protein sequences using GBA. In addition to querying capabilities of the existing GBA algorithm, GBA Manager computes the p-values of the LCR identified. The p-value gives an estimate of the possibility that the region appears by chance. GBA Manager presents the output in three different understandable formats. GBA Manager is freely accessible at http://bioinformatics.cise.ufl.edu/GBA/GBA.htm .

Chromatin looping and eRNA transcription precede the transcriptional activation of gene in the β-globin locus

PubMed Central

Kim, Yea Woon; Lee, Sungkung; Yun, Jangmi; Kim, AeRi

2015-01-01

Enhancers are closely positioned with actively transcribed target genes by chromatin looping. Non-coding RNAs are often transcribed on active enhancers, referred to as eRNAs (enhancer RNAs). To explore the kinetics of enhancer–promoter looping and eRNA transcription during transcriptional activation, we induced the β-globin locus by chemical treatment and analysed cross-linking frequency between the β-globin gene and locus control region (LCR) and the amount of eRNAs transcribed on the LCR in a time course manner. The cross-linking frequency was increased after chemical induction but before the transcriptional activation of gene in the β-globin locus. Transcription of eRNAs was increased in concomitant with the increase in cross-linking frequency. These results show that chromatin looping and eRNA transcription precedes the transcriptional activation of gene. Concomitant occurrence of the two events suggests functional relationship between them. PMID:25588787

Lamprologus markerti, a new lamprologine cichlid (Teleostei: Cichlidae) endemic to the lower Congo River in the Democratic Republic of Congo, west-central Africa.

PubMed

Tougas, Stephanie; Stiassny, Melanie L J

2014-08-15

A new Lamprologus is described from the lower Congo River (LCR) in the Democratic Republic of Congo. Lamprologus markerti, new species, is readily distinguished from L. tigripictilis and L. werneri, the LCR endemic lamprologines with which it was once taxonomically conflated, in the possession of a reduced number of gill rakers on the first arch (9-11 versus 12-17), a longer head (32.1-34.7% SL versus 29.3-31.9 and 29.1-32.9% SL, respectively), and a longer predorsal length (33.0-35.9% SL versus 29.3-32.7 and 28.5-32.6% SL, respectively). Further, L. markerti lacks a second intestinal loop present in both L. tigripictilis and L. werneri, and has a highly reduced infraorbital series often consisting of a single first infraorbital (lachrymal) element. 

Differential structural status of the RNA counterpart of an undecamer quasi-palindromic DNA sequence present in LCR of human β-globin gene cluster.

PubMed

Kaushik, Mahima; Kukreti, Shrikant

2015-01-01

Our previous work on structural polymorphism shown at a single nucleotide polymorphism (SNP) (A → G) site located on HS4 region of locus control region (LCR) of β-globin gene has established a hairpin → duplex equilibrium corresponding to A → B like DNA transition (Kaushik M, Kukreti, R., Grover, D., Brahmachari, S.K. and Kukreti S. Nucleic Acids Res. 2003; Kaushik M, Kukreti S. Nucleic Acids Res. 2006). The G-allele of A → G SNP has been shown to be significantly associated with the occurrence of β-thalassemia. Considering the significance of this 11-nt long quasi-palindromic sequence [5'-TGGGG(G/A)CCCCA; HP(G/A)11] of β-globin gene LCR, we further explored the differential behavior of the same DNA sequence with its RNA counterpart, using various biophysical and biochemical techniques. In contrast to its DNA counterpart exhibiting a A → B structural transition and an equilibrium between duplex and hairpin forms, the studied RNA oligonucleotide sequence [5'-UGGGG(G/A)CCCCA; RHP(G/A)11] existed only in duplex form (A-conformation) and did not form hairpin. The single residue difference from A to G led to the unusual thermal stability of the RNA structure formed by the studied sequence. Since, naturally occurring mutations and various SNP sites may stabilize or destabilize the local DNA/RNA secondary structures, these structural transitions may affect the gene expression by a change in the protein-DNA recognition patterns.

Hepatoprotective potential of Lavandula coronopifolia extracts against ethanol induced oxidative stress-mediated cytotoxicity in HepG2 cells.

PubMed

Farshori, Nida Nayyar; Al-Sheddi, Ebtsam S; Al-Oqail, Mai M; Hassan, Wafaa H B; Al-Khedhairy, Abdulaziz A; Musarrat, Javed; Siddiqui, Maqsood A

2015-08-01

The present investigations were carried out to study the protective potential of four extracts (namely petroleum ether extract (LCR), chloroform extract (LCM), ethyl acetate extract (LCE), and alcoholic extract (LCL)) of Lavandula coronopifolia on oxidative stress-mediated cell death induced by ethanol, a known hepatotoxin in human hapatocellular carcinoma (HepG2) cells. Cells were pretreated with LCR, LCM, LCE, and LCL extracts (10-50 μg/ml) of L. coronopifolia for 24 h and then ethanol was added and incubated further for 24 h. After the exposure, cell viability using (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and neutral red uptake assays and morphological changes in HepG2 cells were studied. Pretreatment with various extracts of L. coronpifolia was found to be significantly effective in countering the cytotoxic responses of ethanol. Antioxidant properties of these L. coronopifolia extracts against reactive oxygen species (ROS) generation, lipid peroxidation (LPO), and glutathione (GSH) levels induced by ethanol were investigated. Results show that pretreatment with these extracts for 24 h significantly inhibited ROS generation and LPO induced and increased the GSH levels reduced by ethanol. The data from the study suggests that LCR, LCM, LCE, and LCL extracts of L. coronopifolia showed hepatoprotective activity against ethanol-induced damage in HepG2 cells. However, a comparative study revealed that the LCE extract was found to be the most effective and LCL the least effective. The hepatoprotective effects observed in the study could be associated with the antioxidant properties of these extracts of L. coronopifolia. © The Author(s) 2013.

Variation in the ovine cortisol response to systemic bacterial endotoxin challenge is predominantly determined by signalling within the hypothalamic-pituitary-adrenal axis

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

You Qiumei; Karrow, Niel A.; Cao Honghe

Bi-directional communication between the neuroendocrine and immune systems is designed, in part, to maintain or restore homeostasis during physiological stress. Exposure to endotoxin during Gram-negative bacterial infection for example, elicits the release of pro-inflammatory cytokines that activate the hypothalamic-pituitary-adrenal axis (HPAA). The secretion of adrenal glucocorticoids subsequently down regulates the host inflammatory response, minimizing potential tissue damage. Sequence and epigenetic variants in genes involved in regulating the neuroendocrine and immune systems are likely to contribute to individual differences in the HPAA response, and this may influence the host anti-inflammatory response to toxin exposure and susceptibility to inflammatory disease. In thismore » study, high (HCR) and low (LCR) cortisol responders were selected from a normal population of 110 female sheep challenged iv with Escherichia coli endotoxin (400 ng/kg) to identify potential determinants that contribute to variation in the cortisol response phenotype. This phenotype was stable over several years in the HCR and LCR animals, and did not appear to be attributed to differences in expression of hepatic immune-related genes or systemic pro-inflammatory cytokine concentrations. Mechanistic studies using corticotrophin-releasing factor (0.5 {mu}g/kg body weight), arginine vasopressin (0.5 {mu}g/kg), and adrenocorticotropic hormone (0.5 {mu}g/kg) administered iv demonstrated that variation in this phenotype is largely determined by signalling within the HPAA. Future studies will use this ovine HCR/LCR model to investigate potential genetic and epigenetic variants that may contribute to variation in cortisol responsiveness to bacterial endotoxin.« less

Erythroid activator NF-E2, TAL1 and KLF1 play roles in forming the LCR HSs in the human adult β-globin locus.

PubMed

Kim, Yea Woon; Yun, Won Ju; Kim, AeRi

2016-06-01

The β-like globin genes are developmental stage specifically transcribed in erythroid cells. The transcription of the β-like globin genes requires erythroid specific activators such as GATA-1, NF-E2, TAL1 and KLF1. However, the roles of these activators have not fully elucidated in transcription of the human adult β-globin gene. Here we employed hybrid MEL cells (MEL/ch11) where a human chromosome containing the β-globin locus is present and the adult β-globin gene is highly transcribed by induction. The roles of erythroid specific activators were analyzed by inhibiting the expression of NF-E2, TAL1 or KLF1 in MEL/ch11 cells. The loss of each activator decreased the transcription of human β-globin gene, locus wide histone hyperacetylation and the binding of other erythroid specific activators including GATA-1, even though not affecting the expression of other activators. Notably, sensitivity to DNase I was reduced in the locus control region (LCR) hypersensitive sites (HSs) with the depletion of activators. These results indicate that NF-E2, TAL1 and KLF1, all activators play a primary role in HSs formation in the LCR. It might contribute to the transcription of human adult β-globin gene by allowing the access of activators and cofactors. The roles of activators in the adult β-globin locus appear to be different from the roles in the early fetal locus. Copyright © 2016 Elsevier Ltd. All rights reserved.

Demystifying Controlling Copper Corrosion

EPA Science Inventory

The LCR systematically misses the highest health and corrosion risk sites for copper. Additionally, there are growing concerns for WWTP copper in sludges and discharge levels. There are many corrosion control differences between copper and lead. This talk explains the sometimes c...

Characterization of dissolved organic matter in drinking water sources impacted by multiple tributaries.

PubMed

Rosario-Ortiz, Fernando L; Snyder, Shane A; Suffet, I H

2007-10-01

The characterization of dissolved organic matter (DOM) in drinking water sources is important as this material contributes to the formation of disinfection by-products (DBPs) and affects how water treatment unit operations are optimized. Drinking water utilities often draw water from sources impacted by multiple tributaries, with possible shifts in DOM concentrations and reactivity over time, depending on specific environmental conditions. In this study, results are presented on the characterization of DOM under varying ambient conditions from the four main tributaries of Lake Mead, a large reservoir in the southwest United States. The tributaries include the Las Vegas Wash (LVW), Muddy River (MR), Virgin River (VR) and the upper Colorado River (UCR). One additional sample was collected at the outflow of the reservoir (lower Colorado River (LCR)). The DOM was characterized by both bulk parameters (specific ultraviolet absorbance (SUVA)) and specific physicochemical properties, i.e. size, polarity and fluorescence. The analyses were performed emphasizing limited changes in its natural configuration by eliminating analytical preparation steps, excluding sample filtration (0.45 microm filter). Results indicate that each tributary had a different molecular weight distribution, as well as fluorescence properties, which helped in the identification of the relative source of DOM (allochthonous versus autochthonous). The largest apparent molecular weight distribution was observed for DOM samples collected at the MR site, which is fed mostly by groundwater seepage. The smallest apparent molecular weight was observed for DOM collected at the LCR site, suggesting that retention in the reservoir resulted in a decrease in molecular weight as a probable result of photo oxidation and microbial processes. Fluorescence analysis aided the differentiation of DOM by clearly identifying waters that were affected by microbial activity (LVW, UCR, and LCR), either by wastewater influence or by autochthonous processes, versus limited microbial influence (MR and VR). Polarity analysis revealed clear differences in the hydrophobic/hydrophilic nature between waters, including temporal differences within individual waters at a particular site. The DOM from the LVW and VR sites had higher hydrophobic character, as measured by retention onto non-polar sorbents. Additionally, the DOM collected at the LCR had the least hydrophobic character. This type of analysis would be beneficial to utilities who want to better understand and manage their source waters, especially in the evaluation of temporal variation within a watershed.

Voluntary Running Aids to Maintain High Body Temperature in Rats Bred for High Aerobic Capacity

PubMed Central

Karvinen, Sira M.; Silvennoinen, Mika; Ma, Hongqiang; Törmäkangas, Timo; Rantalainen, Timo; Rinnankoski-Tuikka, Rita; Lensu, Sanna; Koch, Lauren G.; Britton, Steven L.; Kainulainen, Heikki

2016-01-01

The production of heat, i.e., thermogenesis, is a significant component of the metabolic rate, which in turn affects weight gain and health. Thermogenesis is linked to physical activity (PA) level. However, it is not known whether intrinsic exercise capacity, aging, and long-term voluntary running affect core body temperature. Here we use rat models selectively bred to differ in maximal treadmill endurance running capacity (Low capacity runners, LCR and High capacity Runners, HCR), that as adults are divergent for aerobic exercise capacity, aging, and metabolic disease risk to study the connection between PA and body temperature. Ten high capacity runner (HCR) and ten low capacity runner (LCR) female rats were studied between 9 and 21 months of age. Rectal body temperature of HCR and LCR rats was measured before and after 1-year voluntary running/control intervention to explore the effects of aging and PA. Also, we determined whether injected glucose and spontaneous activity affect the body temperature differently between LCR and HCR rats at 9 vs. 21 months of age. HCRs had on average 1.3°C higher body temperature than LCRs (p < 0.001). Aging decreased the body temperature level of HCRs to similar levels with LCRs. The opportunity to run voluntarily had a significant impact on the body temperature of HCRs (p < 0.001) allowing them to maintain body temperature at a similar level as when at younger age. Compared to LCRs, HCRs were spontaneously more active, had higher relative gastrocnemius muscle mass and higher UCP2, PGC-1α, cyt c, and OXPHOS levels in the skeletal muscle (p < 0.050). These results suggest that higher PA level together with greater relative muscle mass and higher mitochondrial content/function contribute to the accumulation of heat in the HCRs. Interestingly, neither aging nor voluntary training had a significant impact on core body temperature of LCRs. However, glucose injection resulted in a lowering of the body temperature of LCRs (p < 0.050), but not that of HCRs. In conclusion, rats born with high intrinsic capacity for aerobic exercise and better health have higher body temperature compared to rats born with low exercise capacity and disease risk. Voluntary running allowed HCRs to maintain high body temperature during aging, which suggests that high PA level was crucial in maintaining the high body temperature of HCRs. PMID:27504097

Refining the 22q11.2 deletion breakpoints in DiGeorge syndrome by aCGH

PubMed Central

Bittel, D.C.; Yu, S.; Newkirk, H.; Kibiryeva, N.; Holt, S.; Butler, M.G.; Cooley, L.D.

2009-01-01

Hemizygous deletions of the chromosome 22q11.2 region result in the 22q11.2 deletion syndrome also referred to as DiGeorge, Velocardiofacial or Shprintzen syndromes. The phenotype is variable but commonly includes conotruncal cardiac defects, palatal abnormalities, learning and behavioral problems, immune deficiency, and facial anomalies. Four distinct highly homologous blocks of low copy number repeat sequences (LCRs) flank the deletion region. Mispairing of LCRs during meiosis with unequal meiotic exchange is assumed to cause the recurrent and consistent deletions. The proximal LCR is reportedly located at 22q11.2 from 17.037 to 17.083 Mb while the distal LCR is located from 19.835 to 19.880 Mb. Although the chromosome breakpoints are thought to localize to the LCRs, the positions of the breakpoints have been investigated in only a few individuals. Therefore, we used high resolution oligonucleotide-based 244K microarray comparative genomic hybridization (aCGH) to resolve the breakpoints in a cohort of 20 subjects with known 22q11.2 deletions. We also investigated copy number variation (CNV) in the rest of the genome. The 22q11.2 breaks occurred on either side of the LCR in our subjects, although more commonly on the distal side of the reported proximal LCR. The proximal breakpoints in our subjects spanned the region from 17.036 to 17.398 Mb. This region includes the genes DGCR6 (DiGeorge syndrome critical region protein 6) and PRODH (proline dehydrogenase 1), along with three open reading frames that may encode proteins of unknown function. The distal breakpoints spanned the region from 19.788 to 20.122 Mb. This region includes the genes GGT2 (gamma-glutamyltransferase-like protein 2), HIC2 (hypermethylated in cancer 2), and multiple transcripts of unknown function. The genes in these two breakpoint regions are variably hemizygous depending on the location of the breakpoints. Our 20 subjects had 254 CNVs throughout the genome, 94 duplications and 160 deletions, ranging in size from 1 kb to 2.4 Mb. The presence or absence of genes at the breakpoints depending on the size of the deletion plus variation in the rest of the genome due to CNVs likely contribute to the variable phenotype associated with the 22q11.2 deletion or DiGeorge syndrome. PMID:19420922

Voluntary Running Aids to Maintain High Body Temperature in Rats Bred for High Aerobic Capacity.

PubMed

Karvinen, Sira M; Silvennoinen, Mika; Ma, Hongqiang; Törmäkangas, Timo; Rantalainen, Timo; Rinnankoski-Tuikka, Rita; Lensu, Sanna; Koch, Lauren G; Britton, Steven L; Kainulainen, Heikki

2016-01-01

The production of heat, i.e., thermogenesis, is a significant component of the metabolic rate, which in turn affects weight gain and health. Thermogenesis is linked to physical activity (PA) level. However, it is not known whether intrinsic exercise capacity, aging, and long-term voluntary running affect core body temperature. Here we use rat models selectively bred to differ in maximal treadmill endurance running capacity (Low capacity runners, LCR and High capacity Runners, HCR), that as adults are divergent for aerobic exercise capacity, aging, and metabolic disease risk to study the connection between PA and body temperature. Ten high capacity runner (HCR) and ten low capacity runner (LCR) female rats were studied between 9 and 21 months of age. Rectal body temperature of HCR and LCR rats was measured before and after 1-year voluntary running/control intervention to explore the effects of aging and PA. Also, we determined whether injected glucose and spontaneous activity affect the body temperature differently between LCR and HCR rats at 9 vs. 21 months of age. HCRs had on average 1.3°C higher body temperature than LCRs (p < 0.001). Aging decreased the body temperature level of HCRs to similar levels with LCRs. The opportunity to run voluntarily had a significant impact on the body temperature of HCRs (p < 0.001) allowing them to maintain body temperature at a similar level as when at younger age. Compared to LCRs, HCRs were spontaneously more active, had higher relative gastrocnemius muscle mass and higher UCP2, PGC-1α, cyt c, and OXPHOS levels in the skeletal muscle (p < 0.050). These results suggest that higher PA level together with greater relative muscle mass and higher mitochondrial content/function contribute to the accumulation of heat in the HCRs. Interestingly, neither aging nor voluntary training had a significant impact on core body temperature of LCRs. However, glucose injection resulted in a lowering of the body temperature of LCRs (p < 0.050), but not that of HCRs. In conclusion, rats born with high intrinsic capacity for aerobic exercise and better health have higher body temperature compared to rats born with low exercise capacity and disease risk. Voluntary running allowed HCRs to maintain high body temperature during aging, which suggests that high PA level was crucial in maintaining the high body temperature of HCRs.

Best Practices and Simultaneous Compliance

EPA Science Inventory

This wrap-up presentation to the workshop covers several topics concerning how lead and copper compliance under the Lead and Copper Rule should be integrated into an overall “simultaneous compliance” framework with other organizations. The LCR requires “optimization” of lead leve...

Review of a model to assess stranding of juvenile salmon by ship wakes along the Lower Columbia River, Oregon and Washington

USGS Publications Warehouse

Kock, Tobias J.; Plumb, John M.; Adams, Noah S.

2013-01-01

Long period wake waves from deep draft vessels have been shown to strand small fish, particularly juvenile Chinook salmon Oncorhynchus tschawytcha, in the lower Columbia River (LCR). The U.S. Army Corps of Engineers is responsible for maintaining the shipping channel in the LCR and recently conducted dredging operations to deepen the shipping channel from an authorized depth of 40 feet(ft) to an authorized depth of 43 ft (in areas where rapid shoaling was expected, dredging operations were used to increase the channel depth to 48 ft). A model was developed to estimate stranding probabilities for juvenile salmon under the 40- and 43-ft channel scenarios, to determine if channel deepening was going to affect wake stranding (Assessment of potential stranding of juvenile salmon by ship wakes along the Lower Columbia River under scenarios of ship traffic and channel depth: Report prepared for the Portland District U.S. Army Corps of Engineers, Portland, Oregon). The U.S. Army Corps of Engineers funded the U.S. Geological Survey to review this model. A total of 30 review questions were provided to guide the review process, and these questions are addressed in this report. In general, we determined that the analyses by Pearson (2011) were appropriate given the data available. We did identify two areas where additional information could have been provided: (1) a more thorough description of model diagnostics and model selection would have been useful for the reader to better understand the model framework; and (2) model uncertainty should have been explicitly described and reported in the document. Stranding probability estimates between the 40- and 43-ft channel depths were minimally different under most of the scenarios that were examined by Pearson (2011), and a discussion of the effects of uncertainty given these minimal differences would have been useful. Ultimately, however, a stochastic (or simulation) model would provide the best opportunity to illustrate uncertainty within a given set of model predictions, but such an approach would require a substantial amount of additional data collection. Several review questions focused on the accuracy and precision of the model estimates, but we were unable to address these questions because of the limited data that currently exists regarding wake stranding in the LCR. Additional field studies will be required to validate findings from Pearson (2011), if concerns regarding accuracy and precision remain a priority. Although the Pearson (2011) model provided a useful examination of stranding under pre-construction and post-construction conditions, future research will be required to better understand the effects of wake stranding on juvenile salmonids throughout the entire LCR. If additional information on wake stranding is desired in the future, the following topics may be of interest: (1) spatial examination of wake stranding throughout the entire LCR; (2) additional evaluation of juvenile salmonid behavior and population dynamics; (3) assessing and integrating predicted changes in ship development; and (4) assessing and integrating predicted changes in climate on environmental factors known to cause stranding.

Maintaining safe, efficient and sustainable intermodal transport through the Port of Portland.

DOT National Transportation Integrated Search

2011-10-01

"About $15 billion of freight passes annually through the Lower Columbia River (LCR) navigation channel to reach Portland and Vancouver, : where most of it connects with land transport. This commerce plays a vital role in sustaining the regional econ...

LONG-TERM IMPACTS OF ORTHOPHOSPHATE TREATMENT ON COPPER

EPA Science Inventory

Laboratory, pilot, and field data collected support the theoretical "cupric hydroxide" copper solubility model. For the short time frames inherent in laboratory and pilot studies of copper solubility and in initial field monitoring for the LCR from Tier 1 soldered copper sites, c...

GEOSYNTHETIC DESIGN GUIDANCE FOR HAZARDOUS WASTE LANDFILL CELLS AND SURFACE IMPOUNDMENTS

EPA Science Inventory

The report provides guidance design procedures for the use of geosynthetic materials in hazardous waste land disposal cells. Primary geosynthetic components include flexible membrane liners (FML) used to limit the flow of leachate, and leachate collection and removal systems (LCR...

75 FR 81424 - Airworthiness Directives; Sikorsky Aircraft Corporation (Sikorsky) Model S76A, B, and C Helicopters

Federal Register 2010, 2011, 2012, 2013, 2014

2010-12-28

.... The EAD requires inspecting the LITEF Attitude Heading and Reference System (AHRS) unit of the...: Model S-76A, B, and C helicopters, with LITEF LCR-100, Attitude Heading and Reference System (AHRS) Unit...

Evaluation of protective potential of Yersinia pestis outer membrane protein antigens as possible candidates for a new-generation recombinant plague vaccine.

PubMed

Erova, Tatiana E; Rosenzweig, Jason A; Sha, Jian; Suarez, Giovanni; Sierra, Johanna C; Kirtley, Michelle L; van Lier, Christina J; Telepnev, Maxim V; Motin, Vladimir L; Chopra, Ashok K

2013-02-01

Plague caused by Yersinia pestis manifests itself in bubonic, septicemic, and pneumonic forms. Although the U.S. Food and Drug Administration recently approved levofloxacin, there is no approved human vaccine against plague. The capsular antigen F1 and the low-calcium-response V antigen (LcrV) of Y. pestis represent excellent vaccine candidates; however, the inability of the immune responses to F1 and LcrV to provide protection against Y. pestis F1(-) strains or those which harbor variants of LcrV is a significant concern. Here, we show that the passive transfer of hyperimmune sera from rats infected with the plague bacterium and rescued by levofloxacin protected naive animals against pneumonic plague. Furthermore, 10 to 12 protein bands from wild-type (WT) Y. pestis CO92 reacted with the aforementioned hyperimmune sera upon Western blot analysis. Based on mass spectrometric analysis, four of these proteins were identified as attachment invasion locus (Ail/OmpX), plasminogen-activating protease (Pla), outer membrane protein A (OmpA), and F1. The genes encoding these proteins were cloned, and the recombinant proteins purified from Escherichia coli for immunization purposes before challenging mice and rats with either the F1(-) mutant or WT CO92 in bubonic and pneumonic plague models. Although antibodies to Ail and OmpA protected mice against bubonic plague when challenged with the F1(-) CO92 strain, Pla antibodies were protective against pneumonic plague. In the rat model, antibodies to Ail provided protection only against pneumonic plague after WT CO92 challenge. Together, the addition of Y. pestis outer membrane proteins to a new-generation recombinant vaccine could provide protection against a wide variety of Y. pestis strains.

Singlet oxygen explicit dosimetry to predict long-term local tumor control for BPD-mediated photodynamic therapy

NASA Astrophysics Data System (ADS)

Kim, Michele M.; Penjweini, Rozhin; Ong, Yi Hong; Zhu, Timothy C.

2017-02-01

Photodynamic therapy (PDT) is a well-established treatment modality for cancer and other malignant diseases; however, quantities such as light fluence, photosensitizer photobleaching rate, and PDT dose do not fully account for all of the dynamic interactions between the key components involved. In particular, fluence rate (Φ) effects are not accounted for, which has a large effect on the oxygen consumption rate. In this preclinical study, reacted singlet oxygen [1O2]rx was investigated as a dosimetric quantity for PDT outcome. The ability of [1O2]rx to predict the long-term local tumor control rate (LCR) for BPD-mediated PDT was examined. Mice bearing radioactivelyinduced fibrosarcoma (RIF) tumors were treated with different in-air fluences (250, 300, and 350 J/cm2) and in-air ϕ (75, 100, and150 mW/cm2) with a BPD dose of 1 mg/kg and a drug-light interval of 3 hours. Treatment was delivered with a collimated laser beam of 1 cm diameter at 690 nm. Explicit dosimetry of initial tissue oxygen concentration, tissue optical properties, and BPD concentration was used to calculate [1O2]rx. Φ was calculated for the treatment volume based on Monte-Carlo simulations and measured tissue optical properties. Kaplan-Meier analyses for LCR were done for an endpoint of tumor volume <= 100 mm3 using four dose metrics: light fluence, photosensitizer photobleaching rate, PDT dose, and [1O2]rx. PDT dose was defined as the product of the timeintegral of photosensitizer concentration and Φ at a 3 mm tumor depth. Preliminary studies show that [1O2]rx better correlates with LCR and is an effective dosimetric quantity that can predict treatment outcome.

Singlet oxygen explicit dosimetry to predict long-term local tumor control for Photofrin-mediated photodynamic therapy

NASA Astrophysics Data System (ADS)

Penjweini, Rozhin; Kim, Michele M.; Ong, Yi Hong; Zhu, Timothy C.

2017-02-01

Although photodynamic therapy (PDT) is an established modality for the treatment of cancer, current dosimetric quantities do not account for the variations in PDT oxygen consumption for different fluence rates (φ). In this study we examine the efficacy of reacted singlet oxygen concentration ([1O2]rx) to predict long-term local control rate (LCR) for Photofrin-mediated PDT. Radiation-induced fibrosarcoma (RIF) tumors in the right shoulders of female C3H mice are treated with different in-air fluences of 225-540 J/cm2 and in-air fluence rate (φair) of 50 and 75 mW/cm2 at 5 mg/kg Photofrin and a drug-light interval of 24 hours using a 1 cm diameter collimated laser beam at 630 nm wavelength. [1O2]rx is calculated by using a macroscopic model based on explicit dosimetry of Photofrin concentration, tissue optical properties, tissue oxygenation and blood flow changes during PDT. The tumor volume of each mouse is tracked for 90 days after PDT and Kaplan-Meier analyses for LCR are performed based on a tumor volume <=100 mm3, for the four dose metrics light fluence, photosensitizer photobleaching rate, PDT dose and [1O2]rx. PDT dose is defined as a temporal integral of photosensitizer concentration and Φ at a 3 mm tumor depth. φ is calculated throughout the treatment volume based on Monte-Carlo simulation and measured tissue optical properties. Our preliminary studies show that [1O2]rx is the best dosimetric quantity that can predict tumor response and correlate with LCR. Moreover, [1O2]rx calculated using the blood flow changes was in agreement with [1O2]rx calculated based on the actual tissue oxygenation.

Lead in drinking water and human blood lead levels in the United States.

PubMed

Brown, Mary Jean; Margolis, Stephen

2012-08-10

Lead is a pervasive environmental contaminant. The adverse health effects of lead exposure in children and adults are well documented, and no safe blood lead threshold in children has been identified. Lead can be ingested from various sources, including lead paint and house dust contaminated by lead paint, as well as soil, drinking water, and food. The concentration of lead, total amount of lead consumed, and duration of lead exposure influence the severity of health effects. Because lead accumulates in the body, all sources of lead should be controlled or eliminated to prevent childhood lead poisoning. Beginning in the 1970s, lead concentrations in air, tap water, food, dust, and soil began to be substantially reduced, resulting in significantly reduced blood lead levels (BLLs) in children throughout the United States. However, children are still being exposed to lead, and many of these children live in housing built before the 1978 ban on lead-based residential paint. These homes might contain lead paint hazards, as well as drinking water service lines made from lead, lead solder, or plumbing materials that contain lead. Adequate corrosion control reduces the leaching of lead plumbing components or solder into drinking water. The majority of public water utilities are in compliance with the Safe Drinking Water Act Lead and Copper Rule (LCR) of 1991. However, some children are still exposed to lead in drinking water. EPA is reviewing LCR, and additional changes to the rule are expected that will further protect public health. Childhood lead poisoning prevention programs should be made aware of the results of local public water system lead monitoring measurement under LCR and consider drinking water as a potential cause of increased BLLs, especially when other sources of lead exposure are not identified.

Nondestructive testing and characterization of residual stress field using an ultrasonic method

NASA Astrophysics Data System (ADS)

Song, Wentao; Xu, Chunguang; Pan, Qinxue; Song, Jianfeng

2016-03-01

To address the difficulty in testing and calibrating the stress gradient in the depth direction of mechanical components, a new technology of nondestructive testing and characterization of the residual stress gradient field by ultrasonic method is proposed based on acoustoelasticity theory. By carrying out theoretical analysis, the sensitivity coefficients of different types of ultrasonic are obtained by taking the low carbon steel(12%C) as a research object. By fixing the interval distance between sending and receiving transducers, the mathematical expressions of the change of stress and the variation of time are established. To design one sending-one receiving and oblique incidence ultrasonic detection probes, according to Snell law, the critically refracted longitudinal wave (LCR wave) is excited at a certain depth of the fixed distance of the tested components. Then, the relationship between the depth of LCR wave detection and the center frequency of the probe in Q235 steel is obtained through experimental study. To detect the stress gradient in the depth direction, a stress gradient LCR wave detection model is established, through which the stress gradient formula is derived by the relationship between center frequency and detecting depth. A C-shaped stress specimen of Q235 steel is designed to conduct stress loading tests, and the stress is measured with the five group probes at different center frequencies. The accuracy of ultrasonic testing is verified by X-ray stress analyzer. The stress value of each specific depth is calculated using the stress gradient formula. Accordingly, the ultrasonic characterization of residual stress field is realized. Characterization results show that the stress gradient distribution is consistent with the simulation in ANSYS. The new technology can be widely applied in the detection of the residual stress gradient field caused by mechanical processing, such as welding and shot peening.

Evaluation of Protective Potential of Yersinia pestis Outer Membrane Protein Antigens as Possible Candidates for a New-Generation Recombinant Plague Vaccine

PubMed Central

Erova, Tatiana E.; Rosenzweig, Jason A.; Sha, Jian; Suarez, Giovanni; Sierra, Johanna C.; Kirtley, Michelle L.; van Lier, Christina J.; Telepnev, Maxim V.; Motin, Vladimir L.

2013-01-01

Plague caused by Yersinia pestis manifests itself in bubonic, septicemic, and pneumonic forms. Although the U.S. Food and Drug Administration recently approved levofloxacin, there is no approved human vaccine against plague. The capsular antigen F1 and the low-calcium-response V antigen (LcrV) of Y. pestis represent excellent vaccine candidates; however, the inability of the immune responses to F1 and LcrV to provide protection against Y. pestis F1− strains or those which harbor variants of LcrV is a significant concern. Here, we show that the passive transfer of hyperimmune sera from rats infected with the plague bacterium and rescued by levofloxacin protected naive animals against pneumonic plague. Furthermore, 10 to 12 protein bands from wild-type (WT) Y. pestis CO92 reacted with the aforementioned hyperimmune sera upon Western blot analysis. Based on mass spectrometric analysis, four of these proteins were identified as attachment invasion locus (Ail/OmpX), plasminogen-activating protease (Pla), outer membrane protein A (OmpA), and F1. The genes encoding these proteins were cloned, and the recombinant proteins purified from Escherichia coli for immunization purposes before challenging mice and rats with either the F1− mutant or WT CO92 in bubonic and pneumonic plague models. Although antibodies to Ail and OmpA protected mice against bubonic plague when challenged with the F1− CO92 strain, Pla antibodies were protective against pneumonic plague. In the rat model, antibodies to Ail provided protection only against pneumonic plague after WT CO92 challenge. Together, the addition of Y. pestis outer membrane proteins to a new-generation recombinant vaccine could provide protection against a wide variety of Y. pestis strains. PMID:23239803

Production of Recombinant Injectosome and Outer Membrane Proteins from Yersinia Pestis KIM5

DTIC Science & Technology

2009-06-01

Overview ........................................................................ 45 20. Test Ammonium Sulfate Precipitation Overview...52 24. LcrV and YscF Test Expressions .................................................................. 53...coli modify the proteins’ primary structure post - translationally? 1.5 Significance of Results The study of VHH as immunotherapeutics for combating

LONG-TERM IMPACTS OF ORTHOPHOSPHATE TREATMENT ON COPPER LEVELS - PRESENTATION

EPA Science Inventory

Laboratory, pilot, and field data collected support the theoretical "cupric hydroxide" copper solubility model. For the short time frames inherent in laboratory and pilot studies of copper solubility and in initial field monitoring for the LCR from Tier 1 soldered copper sites, c...

Long-Term Effects of Orthophosphate Treatment on Copper Concentration

EPA Science Inventory

Laboratory, pilot, and field data collected support the theoretical “cupric hydroxide” copper solubility model. For the short time frames inherent in laboratory and pilot studies of copper solubility, and in initial field monitoring for the LCR from Tier 1 soldered copper sites,...

LONG-TERM IMPACTS OF ORTHOPHOSPHATE TREATMENT ON COPPER LEVELS

EPA Science Inventory

Laboratory, pilot, and field data collected support the theoretical “cupric hydroxide” copper solubility model. For the short time frames inherent in laboratory and pilot studies of copper solubility and in initial field monitoring for the LCR from Tier 1 soldered copper sites,...

THE SOLUBILITY AND SURFACE CHEMISTRY OF FRESHLY PRECIPITATED COPPER SOLIDS

EPA Science Inventory

Since the implementation of the United States Environmental Protection Agency’s Lead and Copper Rule (LCR) in 1991, a great deal of research has been conducted on copper corrosion and the leaching of copper from materials in drinking water distribution systems. While important...

Detection and Evaluation of Elevated Lead Release from Service Lines: A Field Study

EPA Science Inventory

Comparative stagnation sampling conducted in 32 homes in Chicago, Illinois with lead service (LSLs) demonstrated that the existing regulatory sampling protocol under the U. S. Lead and Copper Rule (LCR) systematically underestimated lead corrosion. Lead levels were highest within...

Disinfection Addition and Disinfection Changes: What It Means to the LCR

EPA Science Inventory

This slide presentation’s general points are: Many protective pipe scales are vey dependent on ORP, and hence, state of disinfection. Adding disinfection to anoxic systems will likely cause big chemistry changes in DS and corrosion. Changing disinfectants could cause major l...

Estimating disperser abundance using open population models that incorporate data from continuous detection PIT arrays

USGS Publications Warehouse

Dzul, Maria C.; Yackulic, Charles B.; Korman, Josh

2017-01-01

Autonomous passive integrated transponder (PIT) tag antenna systems continuously detect individually marked organisms at one or more fixed points over long time periods. Estimating abundance using data from autonomous antennae can be challenging, because these systems do not detect unmarked individuals. Here we pair PIT antennae data from a tributary with mark-recapture sampling data in a mainstem river to estimate the number of fish moving from the mainstem to the tributary. We then use our model to estimate abundance of non-native rainbow trout Oncorhynchus mykiss that move from the Colorado River to the Little Colorado River (LCR), the latter of which is important spawning and rearing habitat for federally-endangered humpback chub Gila cypha. We estimate 226 rainbow trout (95% CI: 127-370) entered the LCR from October 2013-April 2014. We discuss the challenges of incorporating detections from autonomous PIT antenna systems into mark-recapture population models, particularly in regards to using information about spatial location to estimate movement and detection probabilities.

Stress-Triggered Phase Separation Is an Adaptive, Evolutionarily Tuned Response

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

Riback, Joshua A.; Katanski, Christopher D.; Kear-Scott, Jamie L.

In eukaryotic cells, diverse stresses trigger coalescence of RNA-binding proteins into stress granules. In vitro, stress-granule-associated proteins can demix to form liquids, hydrogels, and other assemblies lacking fixed stoichiometry. Observing these phenomena has generally required conditions far removed from physiological stresses. We show that poly(A)-binding protein (Pab1 in yeast), a defining marker of stress granules, phase separates and forms hydrogels in vitro upon exposure to physiological stress conditions. Other RNA-binding proteins depend upon low-complexity regions (LCRs) or RNA for phase separation, whereas Pab1’s LCR is not required for demixing, and RNA inhibits it. Based on unique evolutionary patterns, we createmore » LCR mutations, which systematically tune its biophysical properties and Pab1 phase separation in vitro and in vivo. Mutations that impede phase separation reduce organism fitness during prolonged stress. Poly(A)-binding protein thus acts as a physiological stress sensor, exploiting phase separation to precisely mark stress onset, a broadly generalizable mechanism.« less

Oral immunisation with live aroA attenuated Salmonella enterica serovar Typhimurium expressing the Yersinia pestis V antigen protects mice against plague.

PubMed

Garmory, Helen S; Griffin, Kate F; Brown, Katherine A; Titball, Richard W

2003-06-20

Bubonic and pneumonic plague are caused by the bacterium Yersinia pestis. The V antigen of Y. pestis is a protective antigen against plague. In this study, an aroA attenuated strain of Salmonella enterica serovar Typhimurium (SL3261) has been used to deliver the Y. pestis V antigen as a candidate oral plague vaccine. SL3261 was transformed with the expression plasmid pTrc-LcrV, containing the lcrV gene encoding V antigen. Immunoblot analysis showed V antigen expression in SL3261 in vitro and intragastric immunisation of mice with the recombinant Salmonella resulted in the induction of V antigen-specific serum antibody responses and afforded protection against Y. pestis challenge. However, the antibody responses induced by the recombinant Salmonella did not correlate with the protection afforded, indicating that immune responses other than antibody may play a role in the protection afforded against plague by this candidate vaccine.

Pooling Cervical Swabs and Testing by Ligase Chain Reaction Are Accurate and Cost-Saving Strategies for Diagnosis of Chlamydia trachomatis

PubMed Central

Kapala, J.; Copes, D.; Sproston, A.; Patel, J.; Jang, D.; Petrich, A.; Mahony, J.; Biers, K.; Chernesky, M.

2000-01-01

Specimen pooling to achieve efficiency when testing urine specimens for Chlamydia trachomatis nucleic acids has been suggested. We pooled endocervical swabs from 1,288 women and also tested individual swabs by ligase chain reaction (LCR). Out of 53 positive specimens, pools of 4 or 8 specimens missed two positives, providing 96.2% accuracy compared to individual test results. Dilution and positive-control spiking experiments showed that negative specimens with inhibitors of LCR in the pool reduced the signal. Conversely, two extra positives, detected only through pooling, were negative by individual testing but became positive after storage, suggesting that fresh positive specimens with labile inhibitors may be positive in a pool because of dilution of inhibitors. For this population of women with a 4% prevalence of C. trachomatis infection, substantial savings in cost of reagents (55 to 63%) and technologist time (50 to 63%) made pooling strategies a desirable alternative to individual testing. PMID:10878029

Multiple antigens of Yersinia pestis delivered by live recombinant attenuated Salmonella vaccine strains elicit protective immunity against plague.

PubMed

Sanapala, Shilpa; Rahav, Hannah; Patel, Hetal; Sun, Wei; Curtiss, Roy

2016-05-05

Based on our improved novel Salmonella vaccine delivery platform, we optimized the recombinant attenuated Salmonella typhimurium vaccine (RASV) χ12094 to deliver multiple Yersinia pestis antigens. These included LcrV196 (amino acids, 131-326), Psn encoded on pYA5383 and F1 encoded in the chromosome, their synthesis did not cause adverse effects on bacterial growth. Oral immunization with χ12094(pYA5383) simultaneously stimulated high antibody titers to LcrV, Psn and F1 in mice and presented complete protection against both subcutaneous (s.c.) and intranasal (i.n.) challenges with high lethal doses of Y. pestis CO92. Moreover, no deaths or other disease symptoms were observed in SCID mice orally immunized with χ12094(pYA5383) over a 60-day period. Therefore, the trivalent S. typhimurium-based live vaccine shows promise for a next-generation plague vaccine. Copyright © 2016 Elsevier Ltd. All rights reserved.

Single quantum dot analysis enables multiplexed point mutation detection by gap ligase chain reaction.

PubMed

Song, Yunke; Zhang, Yi; Wang, Tza-Huei

2013-04-08

Gene point mutations present important biomarkers for genetic diseases. However, existing point mutation detection methods suffer from low sensitivity, specificity, and a tedious assay processes. In this report, an assay technology is proposed which combines the outstanding specificity of gap ligase chain reaction (Gap-LCR), the high sensitivity of single-molecule coincidence detection, and the superior optical properties of quantum dots (QDs) for multiplexed detection of point mutations in genomic DNA. Mutant-specific ligation products are generated by Gap-LCR and subsequently captured by QDs to form DNA-QD nanocomplexes that are detected by single-molecule spectroscopy (SMS) through multi-color fluorescence burst coincidence analysis, allowing for multiplexed mutation detection in a separation-free format. The proposed assay is capable of detecting zeptomoles of KRAS codon 12 mutation variants with near 100% specificity. Its high sensitivity allows direct detection of KRAS mutation in crude genomic DNA without PCR pre-amplification. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

LCR-initiated rearrangements at the IDS locus, completed with Alu-mediated recombination or non-homologous end joining.

PubMed

Oshima, Junko; Lee, Jennifer A; Breman, Amy M; Fernandes, Priscilla H; Babovic-Vuksanovic, Dusica; Ward, Patricia A; Wolfe, Lynne A; Eng, Christine M; Del Gaudio, Daniela

2011-07-01

Mucopolysaccharidosis type II (MPS II) is caused by mutations in the IDS gene, which encodes the lysosomal enzyme iduronate-2-sulfatase. In ∼20% of MPS II patients the disorder is caused by gross IDS structural rearrangements. We identified two male cases harboring complex rearrangements involving the IDS gene and the nearby pseudogene, IDSP1, which has been annotated as a low-copy repeat (LCR). In both cases the rearrangement included a partial deletion of IDS and an inverted insertion of the neighboring region. In silico analyses revealed the presence of repetitive elements as well as LCRs at the junctions of rearrangements. Our models illustrate two alternative consequences of rearrangements initiated by non-allelic homologous recombination of LCRs: resolution by a second recombination event (that is, Alu-mediated recombination), or resolution by non-homologous end joining repair. These complex rearrangements have the potential to be recurrent and may be present among those MSP II cases with previously uncharacterized aberrations involving IDS.

Portable water quality monitoring system

NASA Astrophysics Data System (ADS)

Nizar, N. B.; Ong, N. R.; Aziz, M. H. A.; Alcain, J. B.; Haimi, W. M. W. N.; Sauli, Z.

2017-09-01

Portable water quality monitoring system was a developed system that tested varied samples of water by using different sensors and provided the specific readings to the user via short message service (SMS) based on the conditions of the water itself. In this water quality monitoring system, the processing part was based on a microcontroller instead of Lead and Copper Rule (LCR) machines to receive the results. By using four main sensors, this system obtained the readings based on the detection of the sensors, respectively. Therefore, users can receive the readings through SMS because there was a connection between Arduino Uno and GSM Module. This system was designed to be portable so that it would be convenient for users to carry it anywhere and everywhere they wanted to since the processor used is smaller in size compared to the LCR machines. It was also developed to ease the user to monitor and control the water quality. However, the ranges of the sensors' detection still a limitation in this study.

THE SOLUBILITY AND SURFACE CHEMISTRY OF FRESHLY PRECIPITATED COPPER SOLIDS

EPA Science Inventory

Since the implementation of the United States Environmental Protection Agency’s Lead and Copper Rule (LCR) in 1991, a great deal of research has been conducted on copper corrosion and the leaching of copper from materials in drinking water distribution systems. While important p...

CALIFORNIA'S FIRST AERATION PLANTS FOR CORROSION CONTROL

EPA Science Inventory

As required by the Lead and Copper Rule (LCR), Idyllwild Water District (IWD) and Pine Cove Water District (PCWD) conducted a sampling and testing program in 1993-1994. The program began with an evaluation of local household plumbing to identify 20 sites in each District which wo...

LEAD AND COPPER CONTROL: LESSONS LEARNED FROM THE US

EPA Science Inventory

This presentation was invited for the “Inorganic Contaminant Issues in Canada–Research Needs” session, in order to provide an overview of how the assumptions and research behind the US Lead and Copper Rule (LCR) have held up in the past 16 years since promulgati...

Spatial And Temporal Analysis Of Multiple Whitefly Transmitted Virus Infections In Watermelon

USDA-ARS?s Scientific Manuscript database

Squash vein yellowing virus (SqVYV), Cucurbit leaf crumple virus (CuLCrV), and Cucurbit yellow stunting disorder virus (CYSDV) are three whitefly-transmitted viruses recently introduced to Florida that induce visually distinguishable symptoms on watermelon. The epidemiology of these three viruses wa...

USEPA LEAD AND COPPER CORROSION DS RESEARCH AND LCR REGULATORY UPDATE (AWWA WQTC 2006)

EPA Science Inventory

The USEPA is actively addressing lead and copper research needs which continue to develop. The objective of the presentation is to summarize current EPA lead and copper research efforts. Discussion includes project descriptions, a summary of relevant current findings, and discuss...

Computational modeling in the optimization of corrosion control to reduce lead in drinking water

EPA Science Inventory

An international “proof-of-concept” research project (UK, US, CA) will present its findings during this presentation. An established computational modeling system developed in the UK is being calibrated and validated in U.S. and Canadian case studies. It predicts LCR survey resul...

75 FR 9895 - Public Water System Supervision Program Revision for the State of Oklahoma

Federal Register 2010, 2011, 2012, 2013, 2014

2010-03-04

... through Friday, at the following offices: Oklahoma Department of Environmental Quality, Water Quality... ENVIRONMENTAL PROTECTION AGENCY [FRL-9121-9] Public Water System Supervision Program Revision for... Public Water System Supervision Program adopting new regulations for the Lead and Copper Rule (LCR) Short...

77 FR 35676 - Public Water System Supervision Program Revision for the State of Texas

Federal Register 2010, 2011, 2012, 2013, 2014

2012-06-14

... ENVIRONMENTAL PROTECTION AGENCY [FRL-9687-5] Public Water System Supervision Program Revision for... Water System Supervision Program. Texas has adopted the Lead and Copper Rule (LCR) Short-Term Revisions...:30 p.m., Monday through Friday, at the following offices: Texas Commission on Environmental Quality...

Epidemiology and Association of Four Insect-Vectored Viruses in Florida Watermelon

USDA-ARS?s Scientific Manuscript database

Whitefly-transmitted Squash vein yellowing virus (SqVYV), Cucurbit leaf crumple virus (CuLCrV), Cucurbit yellow stunting disorder virus (CYSDV) and aphid-transmitted Papaya ringspot virus type W (PRSV-W) have had serious impact on watermelon production in southwest and west-central Florida in the pa...

Patterns of Virus Distribution in Single and Mixed Infections of Florida Watermelons

USDA-ARS?s Scientific Manuscript database

Whitefly-transmitted Squash vein yellowing virus (SqVYV) and Cucurbit leaf crumple virus (CuLCrV), and aphid-transmitted Papaya ringspot virus type W (PRSV-W) have had serious impact on watermelon production in southwest and west-central Florida in recent years. Tissue blot nucleic acid hybridizati...

Investigations of Crown Gall in the Commercial Propagation of Weeping Fig

USDA-ARS?s Scientific Manuscript database

Whitefly-transmitted Squash vein yellowing virus (SqVYV) and Cucurbit leaf crumple virus (CuLCrV), and aphid-transmitted Papaya ringspot virus type W (PRSV-W) have had serious impact on watermelon production in southwest and west-central Florida in recent years. Tissue blot nucleic acid hybridizati...

A Field Comparison of Sampling Protocols for Measuring Lead in Drinking Water

EPA Science Inventory

US EPA Region 5 conducted a sampling study that demonstrates existing sampling protocols used for the Lead and Copper Rule (LCR) underestimate peak and probable mass of lead released in a system with lead service lines (LSLs). This comparative stagnation sampling was conducted i...

Effects of Water Quality and Orthophosphate on Surface Characteristics of Cu Corrosion in Drinking Water using Atomic Force Microscopy

EPA Science Inventory

Since the passage of the Lead and Copper Rule (LCR) in 1991, researchers have examined the effects of water chemistry on the solubility of copper to establish best approaches for reducing copper levels. Despite recent developments, important gaps still exist regarding copper cor...

Effect of variable annual precipitation and nutrient input on nitrogen and phosphorus transport from two Midwestern agricultural watersheds

USDA-ARS?s Scientific Manuscript database

Precipitation patterns and nutrient inputs impact transport of nitrate (NO3-N) and phosphorus (TP) from Midwest watersheds. Nutrient concentrations and yields from two subsurface-drained watersheds, the Little Cobb River (LCR) in southern Minnesota and the South Fork Iowa River (SFIR) in northern Io...

75 FR 15499 - National Primary Drinking Water Regulations; Announcement of the Results of EPA's Review of...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-03-29

...--packed tower aeration PWS--public water system R2S2--Regulatory Review Support Spreadsheet RED...--hexavalent chromium CWS--community water system DBPs--disinfection byproducts DBCP--1,2-dibromo-3... Eligibility Decision IRIS--Integrated Risk Information System LCR--Lead and Copper Rule LH--lutenizing hormone...

Distribution of Four Viruses in Single and Mixed Infections Within Infected Watermelon Plants in Florida

USDA-ARS?s Scientific Manuscript database

Whitefly-transmitted Squash vein yellowing virus (SqVYV) and Cucurbit leaf crumple virus (CuLCrV), and aphid-transmitted Papaya ringspot virus type W (PRSV-W) have had serious impact on watermelon production in southwest and west-central Florida in recent years. Tissue blot nucleic acid hybridizati...

76 FR 67187 - National Drinking Water Advisory Council; Notice of a Public Teleconference Meeting

Federal Register 2010, 2011, 2012, 2013, 2014

2011-10-31

... requirements of the National Drinking Water Regulations for Lead and Copper. DATES: The public teleconference... and Copper: EPA is developing proposed revisions to the Lead and Copper Rule (LCR), which is the National Primary Drinking Water Regulation for controlling lead and copper in drinking water supplied by...

Investigation of the Preservation Method within Environmental Protection Agency Method 200.8

EPA Science Inventory

Lead (Pb) is a trace metal that is closely regulated in drinking water systems because of its harmful toxicity. The U.S. Environmental Protection Agency (USEPA) issued the Lead and Copper Rule (LCR), which defines the action level for Lead as 0.015 mg/L. Researchers and drinking ...

75 FR 63177 - Notice of a Public Meeting: Stakeholder Meeting Concerning EPA's Long-Term Revisions to the...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-10-14

... ENVIRONMENTAL PROTECTION AGENCY [FRL-9213-1] Notice of a Public Meeting: Stakeholder Meeting...: Environmental Protection Agency (EPA). ACTION: Notice. SUMMARY: The U.S. Environmental Protection Agency (EPA... Copper Rule (LCR). EPA is holding a stakeholder meeting to provide information to the public and an...

34 CFR 222.41 - How does a State educational agency compute local contribution rates based upon generally...

Code of Federal Regulations, 2010 CFR

2010-07-01

..., DEPARTMENT OF EDUCATION IMPACT AID PROGRAMS Payments for Federally Connected Children Under Section 8003(b..., subject to the Secretary's review and approval, shall compute an LCR for each group of generally... of this section, the SEA shall consider only those aggregate current expenditures made by the...

76 FR 1392 - Endangered and Threatened Species: Designation of Critical Habitat for Threatened Lower Columbia...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-01-10

... Columbia River. Although there is some level of reproductive isolation and ecological specialization... isolation of the remaining naturally produced fish confer considerable risks to LCR coho. Major habitat... changes are warranted, we will document the bases for the revisions and include this rationale as part of...

Patterns of Virus Distribution in Single and Mixed Infections of Florida Watermelons

USDA-ARS?s Scientific Manuscript database

Whitefly-transmitted Squash vein yellowing virus (SqVYV) and Cucurbit leaf crumple virus (CuLCrV), and aphid-transmitted Papaya ringspot virus type W (PRSV-W) have had serious impact on watermelon production in southwest and west-central Florida in recent years. To determine the distribution of vir...

Epidemiological Analysis of Multi-Virus Infections of Watermelon in Experimental Fields in Southwest Florida

USDA-ARS?s Scientific Manuscript database

Whitefly-transmitted Squash vein yellowing virus (SqVYV) and Cucurbit leaf crumple virus (CuLCrV), and aphid-transmitted Papaya ringspot virus type W (PRSV-W) have had serious impact on watermelon production in southwest and west-central Florida in recent years. Tissue blot nucleic acid hybridizati...

Importance of Fe and Mn Pipe Deposits to Lead and Copper Rule Compliance - abstract

EPA Science Inventory

When Madison, WI exceeded the lead Action Level in 1992, residential and off-line tests suggested that lead release into the water was more complex than a lead solubility mechanism. The water utility chose to address the Lead and Copper Rule (LCR) exceedance by implementing full ...

Incorporating temporal heterogeneity in environmental conditions into a somatic growth model

USGS Publications Warehouse

Dzul, Maria C.; Yackulic, Charles B.; Korman, Josh; Yard, Michael D.; Muehlbauer, Jeffrey D.

2017-01-01

Evaluating environmental effects on fish growth can be challenging because environmental conditions may vary at relatively fine temporal scales compared to sampling occasions. Here we develop a Bayesian state-space growth model to evaluate effects of monthly environmental data on growth of fish that are observed less frequently (e.g., from mark-recapture data where time between captures can range from months to years). We assess effects of temperature, turbidity duration, food availability, flow variability, and trout abundance on subadult humpback chub (Gila cypha) growth in two rivers, the Colorado River (CR) and the Little Colorado River (LCR), and we use out-of-sample prediction to rank competing models. Environmental covariates explained a high proportion of the variation in growth in both rivers; however, the best growth models were river-specific and included either positive temperature and turbidity duration effects (CR) or positive temperature and food availability effects (LCR). Our approach to analyzing environmental controls on growth should be applicable in other systems where environmental data vary over relatively short time scales compared to animal observations.

Human papillomavirus genotypes and phylogenetic analysis of HPV-16 variants in HIV-1 infected subjects in Italy.

PubMed

Tanzi, Elisabetta; Amendola, Antonella; Bianchi, Silvia; Fasolo, M Michela; Beretta, Rosangela; Pariani, Elena; Zappa, Alessandra; Frati, Elena; Orlando, Giovanna

2009-05-29

A cross-sectional study was carried out to improve the state of evidence regarding the spectrum of HPV types and HPV-16 LCR variants circulating among men and women infected with HIV-1 in Italy. This study, conducted in 518 HIV-positive subjects (346 males and 172 females), showed a high prevalence of HPV anal infections (88.7%) in men and of cervical infections (65.1%) in women. A wide spectrum of HPV genotypes has been observed, as both single and multiple infections. Low-risk HPV types 6, 11 and 61 were frequently detected. HPV-16 was the prevalent high-risk type. Fourteen different HPV-16 LCR variants were found. Ten belonged to the European lineage (78.7% were detected in Italian subjects and 21.3% in foreign-born, all homo/bisexual men), two to the Asiatic lineage and two to the African-2 lineage. This study underlines the great genotypic heterogeneity characterizing anal and cervical HPV infections and the marked polymorphism of the predominant HPV-16 in this high-risk population in Italy.

Chlorogenic acid, anthocyanin and flavan-3-ol biosynthesis in flesh and skin of Andean potato tubers (Solanum tuberosum subsp. andigena).

PubMed

Valiñas, Matías Ariel; Lanteri, María Luciana; Ten Have, Arjen; Andreu, Adriana Balbina

2017-08-15

Natural variation of Andean potato was used to study the biosynthesis of phenolic compounds. Levels of phenolic compounds and corresponding structural gene transcripts were examined in flesh and skin of tubers. Phenolic acids, mainly chlorogenic acid (CGA), represent the major compounds, followed by anthocyanins and flavan-3-ols. High-anthocyanin varieties have high levels of CGA. Both metabolite and transcript levels were higher in skin than in flesh and showed a good correspondence. Two hydroxycinnamoyl-CoA transferases (HCT/HQT) have been involved in CGA production, of which HCT reflects CGA levels. Catechin was found in pigmented tissues whereas epicatechin was restricted to tuber skin. Transcripts of leucoanthocyanidin reductase (LCR), which generates catechin, could not be detected. Anthocyanidin reductase (ANR) transcripts, the enzyme responsible for epicatechin production, showed similar levels among samples. These data suggest that the biosynthesis of flavan-3-ols in potato tuber would require ANR but not LCR and that an epimerization process is involved. Copyright © 2017 Elsevier Ltd. All rights reserved.

Proteomic and biochemical basis for enhanced growth yield of Enterobacter sp. LCR1 on insoluble phosphate medium.

PubMed

Kumar, Arvind; Rai, Lal Chand

2015-01-01

Proteomics and biochemical analyses were used to unravel the basis for higher growth yield of Enterobacter sp. LCR1 on insoluble phosphate medium compared to soluble. Proteomic analysis using 2-DE, MALDI-TOF/MS and LC-MS revealed the involvement of nine proteins. Down-regulation of fructose bisphosphate aldolase with decreased concentrations of glucose-6-phosphate and fructose-6-phosphate indicated diminished glycolysis. However, up-regulation of phosphoglycerate mutase, increase in the activities of 6-phosphogluconate dehydratase, 2-keto-3-deoxy-6-phosphogluconate aldolase and 6-phosphogluconate dehydrogenase suggested induction of Entner-Doudoroff and pentose phosphate pathways. These pathways generate sufficient energy from gluconic acid, which is also used for biosynthesis as indicated by up-regulation of elongation factor Tu, elongation factor G and protein disulfide isomerase. Increased reactive oxygen species (ROS) formation resulting from organic acid oxidation leads to overexpressed manganese superoxide dismutase and increased activities of catalase and ascorbate peroxidase. Thus the organism uses gluconate instead of glucose for energy, while alleviating extra ROS formation by oxidative defense enzymes. Copyright © 2014 Elsevier GmbH. All rights reserved.

The 3’-Jα Region of the TCRα Locus Bears Gene Regulatory Activity in Thymic and Peripheral T Cells

PubMed Central

Kučerová-Levisohn, Martina; Knirr, Stefan; Mejia, Rosa I.; Ortiz, Benjamin D.

2015-01-01

Much progress has been made in understanding the important cis-mediated controls on mouse TCRα gene function, including identification of the Eα enhancer and TCRα locus control region (LCR). Nevertheless, previous data have suggested that other cis-regulatory elements may reside in the locus outside of the Eα/LCR. Based on prior findings, we hypothesized the existence of gene regulatory elements in a 3.9-kb region 5’ of the Cα exons. Using DNase hypersensitivity assays and TCRα BAC reporter transgenes in mice, we detected gene regulatory activity within this 3.9-kb region. This region is active in both thymic and peripheral T cells, and selectively affects upstream, but not downstream, gene expression. Together, these data indicate the existence of a novel cis-acting regulatory complex that contributes to TCRα transgene expression in vivo. The active chromatin sites we discovered within this region would remain in the locus after TCRα gene rearrangement, and thus may contribute to endogenous TCRα gene activity, particularly in peripheral T cells, where the Eα element has been found to be inactive. PMID:26177549

Estimative of conversion fractions of AGN magnetic luminosity to produce ultra high energy cosmic rays from the observation of Fermi-LAT gamma rays

NASA Astrophysics Data System (ADS)

Coimbra-Araújo, Carlos H.; Anjos, Rita C.

2017-01-01

A fraction of the magnetic luminosity (LB) produced by Kerr black holes in some active galactic nuclei (AGNs) can produce the necessary energy to accelerate ultra high energy cosmic rays (UHECRs) beyond the GZK limit, observed, e.g., by the Pierre Auger experiment. Nevertheless, the direct detection of those UHECRs has a lack of information about the direction of the source from where those cosmic rays are coming, since charged particles are deflected by the intergalactic magnetic field. This problem arises the needing of alternative methods to evaluate the luminosity of UHECRs (LCR) from a given source. Methods proposed in literature range from the observation of upper limits in gamma rays to the observation of upper limits in neutrinos produced by cascade effects during the propagation of UHECRs. In this aspect, the present work proposes a method to calculate limits of the main possible conversion fractions ηCR = LCR/LB for nine UHECR AGN Seyfert sources based on the respective observation of gamma ray upper limits from Fermi-LAT data.

Appearance of the pituitary factor Pit-1 increases chromatin remodeling at hypersensitive site III in the human GH locus.

PubMed

Yang, Xiaoyang; Jin, Yan; Cattini, Peter A

2010-07-01

Expression of pituitary and placental members of the human GH and chorionic somatomammotropin (CS) gene family is directed by an upstream remote locus control region (LCR). Pituitary-specific expression of GH requires direct binding of Pit-1 (listed as POU1F1 in the HUGO database) to sequences marked by a hypersensitive site (HS) region (HS I/II) 14.6 kb upstream of the GH-N gene (listed as GH1 in the HUGO database). We used human embryonic kidney 293 (HEK293) cells overexpressing wild-type and mutant Pit-1 proteins as a model system to gain insight into the mechanism by which Pit-1 gains access to the GH LCR. Addition of Pit-1 to these cells increased DNA accessibility at HS III, located 28 kb upstream of the human GH-N gene, in a POU homeodomain-dependent manner, as reflected by effects on histone hyperacetylation and RNA polymerase II activity. Direct binding of Pit-1 to HS III sequences is not supported. However, the potential for binding of ETS family members to this region has been demonstrated, and Pit-1 association with this ETS element in HS III sequences requires the POU homeodomain. Also, both ETS1 and ELK1 co-precipitate from human pituitary extracts using two independent sources of Pit-1 antibodies. Finally, overexpression of ELK1 or Pit-1 expression in HEK293 cells increased GH-N RNA levels. However, while ELK1 overexpression also stimulated placental CS RNA levels, the effect of Pit-1 appeared to correlate with ETS factor levels and target GH-N preferentially. These data are consistent with recruitment and an early role for Pit-1 in remodeling of the GH LCR at the constitutively open HS III through protein-protein interaction.

Clinical application of computed tomography-guided (125)I seed interstitial implantation for head and neck cancer patients with unmanageable cervical lymph node metastases.

PubMed

Huang, Hai; Xu, Shaonian; Li, Fusheng; Du, Zhenguang; Wang, Liang

2016-04-27

To assess clinical application of computed tomography (CT)-guided (125)I seed implantation for patients who cannot endure or unwillingly receive repeated surgery, chemotherapy, or radiotherapy for unmanageable cervical lymph node metastases in head and neck cancer (HNC). Thirty-one consecutive patients received CT-guided (125)I seed implantation between February 2010 and December 2013. To evaluate the clinical efficiency, karnofsky performance score (KPS), numeric rating scale (NRS), and tumor volume at 3-, and 6-month post-implantation were compared with pre-implantation, along with local control rate (LCR), overall survival rate (OSR), and complications at 3, 6 months, 1, and 2 years. The tumor volume was obviously decreased at 3-, and 6-month post-implantation (21.23 ± 8.83 versus 9.19 ± 7.52 cm(2); 21.23 ± 8.83 versus 6.42 ± 9.79 cm(2); P < 0.05) compared with pre-implantation. The NRS was statistically reduced (3.06 ± 1.06 versus 7.77 ± 0.92; 2.39 ± 1.15 versus 7.77 ± 0.92; P < 0.05), while KPS was significantly improved (83.18 ± 5.97 versus 73.60 ± 7.90; 82.86 ± 5.43 versus 73.60 ± 7.90; P < 0.05) postoperatively at 3 and 6 months, respectively. The LCR at 3, 6 months, 1, and 2 years was 96.30, 83.87, 64.51, and 45.16%, respectively. The OSR was 100, 100, 67.74, and 45.16%, respectively. Three cases experienced grade I and two had grade II acute radiation toxicity. CT-guided seed implantation may be feasible and safe for HNC patients whose neck nodes are not manageable by routine strategies with fewer complications, higher LCR, and significant pain relief.

Phylogeny and polymorphism in the long control regions E6, E7, and L1 of HPV Type 56 in women from southwest China

PubMed Central

Jing, Yaling; Wang, Tao; Chen, Zuyi; Ding, Xianping; Xu, Jianju; Mu, Xuemei; Cao, Man; Chen, Honghan

2018-01-01

Globally, human papillomavirus (HPV)-56 accounts for a small proportion of all high-risk HPV types; however, HPV-56 is detected at a higher rate in Asia, particularly in southwest China. The present study analyzed polymorphisms, intratypic variants, and genetic variability in the long control regions (LCR), E6, E7, and L1 of HPV-56 (n=75). The LCRs, E6, E7 and L1 were sequenced using a polymerase chain reaction and the sequences were submitted to GenBank. Maximum-likelihood trees were constructed using Kimura's two-parameter model, followed by secondary structure analysis and protein damaging prediction. Additionally, in order to assess the effect of variations in the LCR on putative binding sites for cellular proteins, MATCH server was used. Finally, the selection pressures of the E6-E7 and L1 genes were estimated. A total of 18 point substitutions, a 42-bp deletion and a 19-bp deletion of LCR were identified. Some of those mutations are embedded in the putative binding sites for transcription factors. 18 single nucleotide changes occurred in the E6-E7 sequence, 11/18 were non-synonymous substitutions and 7/18 were synonymous mutations. A total 24 single nucleotide changes were identified in the L1 sequence, 6/24 being non-synonymous mutations and 18/24 synonymous mutations. Selective pressure analysis predicted that the majority of mutations of HPV-56 E6, E7 and L1 were of positive selection. The phylogenetic tree demonstrated that the isolates distributed in two lineages. Data on the prevalence and genetic variation of HPV-56 types in southwest China may aid future studies on viral molecular mechanisms and contribute to future investigations of diagnostic probes and therapeutic vaccines. PMID:29568922

The Impact of Water Quality, Corrosion Inhibitors and Plumbing Age on Copper Release in Distribution Systems

EPA Science Inventory

The U.S. Environmental Protection Agency promulgated the Lead and Copper Rule (LCR) in 1991, which established a copper action level of 1.3 mg/L in the consumers’ tap water. As a result, researchers have examined the effects of water chemistry on the solubility of copper to esta...

34 CFR 222.39 - How does a State educational agency identify generally comparable local educational agencies for...

Code of Federal Regulations, 2012 CFR

2012-07-01

..., DEPARTMENT OF EDUCATION IMPACT AID PROGRAMS Payments for Federally Connected Children Under Section 8003(b... location factors, the SEA shall subdivide according to the size factor before the location factor. (b... follows: (1) The SEA shall not, when computing an LCR, include the following “significantly impacted” LEAs...

34 CFR 222.39 - How does a State educational agency identify generally comparable local educational agencies for...

Code of Federal Regulations, 2014 CFR

2014-07-01

..., DEPARTMENT OF EDUCATION IMPACT AID PROGRAMS Payments for Federally Connected Children Under Section 8003(b... location factors, the SEA shall subdivide according to the size factor before the location factor. (b... follows: (1) The SEA shall not, when computing an LCR, include the following “significantly impacted” LEAs...

Gut microbiota are linked to increased susceptibility to hepatic steatosis in low aerobic capacity rats fed an acute high fat diet

USDA-ARS?s Scientific Manuscript database

Poor aerobic fitness is linked to nonalcoholic fatty liver disease and increased all-cause mortality. We previously found that low capacity running (LCR) rats fed acute high fat diet (HFD; 45% kcal from fat) for 3 days resulted in positive energy balance and increased hepatic steatosis compared with...

Update on the watermelon vine decline virus and other whitefly-transmitted cucurbit viruses in Florida, and their effects on watermelon

USDA-ARS?s Scientific Manuscript database

Whitefly-transmitted Squash vein yellowing virus (SqVYV) was shown in the mid-2000’s to cause a watermelon vine decline in southwest and west-central Florida. More recently, Cucurbit leaf crumple virus (CuLCrV) and Cucurbit yellow stunting disorder virus (CYSDV), also whitefly-transmitted, have bee...

Internal Stress Monitoring of In-Service Structural Steel Members with Ultrasonic Method

PubMed Central

Li, Zuohua; He, Jingbo; Teng, Jun; Wang, Ying

2016-01-01

Internal stress in structural steel members is an important parameter for steel structures in their design, construction, and service stages. However, it is hard to measure via traditional approaches. Among the existing non-destructive testing (NDT) methods, the ultrasonic method has received the most research attention. Longitudinal critically refracted (Lcr) waves, which propagate parallel to the surface of the material within an effective depth, have shown great potential as an effective stress measurement approach. This paper presents a systematic non-destructive evaluation method to determine the internal stress in in-service structural steel members using Lcr waves. Based on theory of acoustoelasticity, a stress evaluation formula is derived. Factor of stress to acoustic time difference is used to describe the relationship between stress and measurable acoustic results. A testing facility is developed and used to demonstrate the performance of the proposed method. Two steel members are measured by using the proposed method and the traditional strain gauge method for verification. Parametric studies are performed on three steel members and the aluminum plate to investigate the factors that influence the testing results. The results show that the proposed method is effective and accurate for determining stress in in-service structural steel members. PMID:28773347

Internal Stress Monitoring of In-Service Structural Steel Members with Ultrasonic Method.

PubMed

Li, Zuohua; He, Jingbo; Teng, Jun; Wang, Ying

2016-03-23

Internal stress in structural steel members is an important parameter for steel structures in their design, construction, and service stages. However, it is hard to measure via traditional approaches. Among the existing non-destructive testing (NDT) methods, the ultrasonic method has received the most research attention. Longitudinal critically refracted (Lcr) waves, which propagate parallel to the surface of the material within an effective depth, have shown great potential as an effective stress measurement approach. This paper presents a systematic non-destructive evaluation method to determine the internal stress in in-service structural steel members using Lcr waves. Based on theory of acoustoelasticity, a stress evaluation formula is derived. Factor of stress to acoustic time difference is used to describe the relationship between stress and measurable acoustic results. A testing facility is developed and used to demonstrate the performance of the proposed method. Two steel members are measured by using the proposed method and the traditional strain gauge method for verification. Parametric studies are performed on three steel members and the aluminum plate to investigate the factors that influence the testing results. The results show that the proposed method is effective and accurate for determining stress in in-service structural steel members.

In Vivo Photo-Cross-Linking to Study T3S Interactions Demonstrated Using the Yersinia pestis T3S System.

PubMed

Henderson, Thomas A; Nilles, Matthew L

2017-01-01

Cross-linking of proteins is effective in determining protein-protein interactions. The use of photo-cross-linkers was developed to study protein interactions in several manners. One method involved the incorporation of photo-activatable cross-linking groups into chemically synthesized peptides. A second approach relies on incorporation of photo-activatable cross-linking groups into proteins using tRNAs with chemically bound photo-activatable amino acids with suppressor tRNAs translational systems to incorporate the tags into specific sites. A third system was made possible by the development of photoreactive amino acids that use the normal cellular tRNAs and aminoacyl tRNA synthetases. In this method, the third system is used to demonstrate its utility for the study of T3S system interactions. This method describes how two photo-activatable amino acids, photo-methionine and photo-leucine, that use the normal cellular machinery are incorporated into Yersinia pestis and used to study interactions in the T3S system. To demonstrate the system, the method was used to cross-link the T3S regulatory proteins LcrG and LcrV.

Biophysical Characterization of the Type III Secretion Tip Proteins and the Tip Proteins Attached to Bacterium-Like Particles

PubMed Central

Choudhari, Shyamal P.; Chen, Xiaotong; Kim, Jae Hyun; van Roosmalen, Maarten L.; Greenwood, Jamie C.; Joshi, Sangeeta B.; Picking, William D.; Leenhouts, Kees; Middaugh, C. Russell; Picking, Wendy L.

2014-01-01

Bacterium-like particles (BLPs), derived from Lactococcus lactis, offer a self-adjuvanting delivery vehicle for subunit protein vaccines. Proteins can be specifically loaded onto the BLPs via a peptidoglycan anchoring domain (PA). In this study, the tip proteins IpaD, SipD and LcrV belonging to type three secretion systems of Shigella flexneri, Salmonella enterica and Yersinia enterocolitica, respectively, were fused to the PA and loaded onto the BLPs. Herein, we biophysically characterized these nine samples and condensed the spectroscopic results into three-index empirical phase diagrams (EPDs). The EPDs show distinctions between the IpaD/SipD and LcrV subfamilies of tip proteins, based on their physical stability, even upon addition of the PA. Upon attachment to the BLPs, the BLPs become defining moiety in the spectroscopic measurements, leaving the tip proteins to have a subtle yet modulating effect on the structural integrity of the tip proteins-BLPs binding. In summary, this work provides a comprehensive view of physical stability of the tip proteins and tip protein-BLPs and serves as a baseline for screening of excipients to increase the stability of the tip protein-BLPs for future vaccine formulation. PMID:24916512

Characterization of mechanical properties and electrochemical behaviour in a Hank´s solution of 316L/Cr1- xAlxN system

NASA Astrophysics Data System (ADS)

Osorio, D. M.; Caicedo, J. C.; Aperador, W.; Benitez-Castro, A. M.; Giraldo-Betancur, A. L.; Muñoz-Saldaña, J.; Yañez-Limón, J. M.; Sanchez, O.; Zambrano, G.

2017-01-01

Cr1-xAlxN hard coatings were successfully deposited by R.F. reactive magnetron co-sputtering in an Ar/N2 gas mixture using chromium and aluminium targets on 316L stainless steel substrates. Crystallographic orientations associated to the Cr1-xAlxN FCC based in the conjugate complex of CrN and w-AlN phases, with ao=4.18Å lattice parameter for the ternary Cr1-xAlxN compound were identified by X-Ray diffraction. The thickness and roughness of the deposited coatings are 1.00±0.05nm and 2.65±0.6nm, respectively. The mechanical properties were determined by nanoindentation leading to a hardness of 27.8±2.6GPa and elastic modulus of 346GPa. The corrosion resistance of the coated 316L/Cr1-xAlxN system under simulated body fluid (SBF, Hank’s solution) was determined via electrochemical impedance spectroscopy. A reduction in the corrosion rate of 99% in relation to uncoated 316L stainless steel substrate was found by Tafel. Thus, these coatings seem to be excellent candidates to be used in biomedical applications.

New insights into ocean tide loading corrections on tidal gravity data in Canary Islands

NASA Astrophysics Data System (ADS)

Arnoso, J.; Benavent, M.; Bos, M. S.; Montesinos, F. G.

2009-04-01

The Canary Islands are an interesting area to investigate ocean tides loading effects due to the complex coastline of the islands and the varying bathymetry. We present here the quality of five recent global oceanic tidal models, GOT00.2, GOT4.7, FES2004, TPXO.7.1 and AG2006, by comparing their predicted ocean tide loading values with results from tidal gravity observations made on three islands, Lanzarote, Tenerife and El Hierro, for the four harmonic constituents O1, K1, M2 and S2. In order to improve the accuracy of the loading corrections on the gravity tide measurements, we have used the high resolution regional oceanic model CIAM2 to supplement the global models considered here. This regional model has been obtained by assimilating TOPEX/Poseidon altimetry at crossovers and along-track points and tide gauge observations into a hydrodynamic model. The model has a 5'Ã-5' resolution and covers the area between the coordinates 26°.5N to 30°.0N and 19°.0W to 12°.5W. The gravity tide observing sites have been occupied by three different LaCoste&Romberg (LCR) spring gravimeters during different periods of observation. We considered here the most recent gravity tide observations made with LCR Graviton-EG1194 in El Hierro Island, for a period of 6 months during 2008. In the case of Tenerife and Lanzarote sites we have used observation periods of 6 months and 8 years with LCR-G665 and LCR-G434 gravimeters, respectively. The last two sites have been revisited in order to improve the previous tidal analysis results. Thus, the gravity ocean tide loading corrections, based on the five global ocean tide models supplemented with the regional model CIAM2 allowed us to review the normalization factors (scale factor and phase lag) of both two gravimeters. Also, we investigated the discrepancies of the corrected gravimetric factors with the DDW elastic and inelastic non hydrostatic body tide model (Dehant et al., 1999). The lowest values are found for inelastic model in the case of M2 and O1 waves at three sites. However, the scatter between oceanic models seen at final residual vectors does not indicate clearly if tidal observations are close to elastic or inelastic body tide model. Finally, after computing misfits of gravity tide observations and ocean tide loading calculations the level of agreement between the five global oceanic models is below 0.2 Gal (1 Gal=10-8ms-2), except for the solar harmonic K1, which reaches a large value that reflects the thermal instability at three sites because the period of K1 is very close to that of S1. None of the five global models seems to give results that are clearly better than the other models.

Effectiveness of the Preservation Protocol within the Environmental Protection Agency (EPA) Method 200.8 for Soluble and Particulate Lead Recovery in Drinking Water

EPA Science Inventory

Lead (Pb) is a toxic trace metal that is regulated in drinking water. The U.S. Environmental Protection Agency (USEPA) issued the Lead and Copper Rule (LCR), which defines the action level for lead at the tap as 0.015 mg/L. Researchers and drinking water utilities typically emplo...

SU-E-I-02: Characterizing Low-Contrast Resolution for Non-Circular CBCT Trajectories

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

Davis, A; Pan, X; Pelizzari, C

Purpose: The use of non-circular scanning trajectories with optimization-basedreconstruction algorithms can be used in conjunction with non-planaracquisition geometries for axial field-of-view (FOV) extension incone-beam CT (CBCT). To evaluate the utility of these trajectories,quantitative image quality metrics should be evaluated. Low-contrastresolution (LCR) and CT number accuracy are significant challenges forCBCT. With unprecedented axial coverage provided by thesetrajectories, measuring such metrics throughout the axial range iscritical. There are currently no phantoms designed to measurelow-contrast resolution over such an extended volume. Methods: The CATPHAN (The Phantom Laboratory, Salem NY) is the current standardfor image quality evaluation. While providing several useful modulesfor different evaluationmore » metrics, each module was designed to beevaluated in a single slice and not for comparison across axialpositions. To characterize the LCR and HU accuracy over an extendedaxial length, we have designed and built a phantom with evaluationmodules at multiple and adjustable axial positions. Results: The modules were made from a cast polyurethane resin. Holes rangingfrom 1/8 to 5/8 inch were added at a constant radius from the modulecenter into which rods of two different plastic materials were pressedto provide two nominal levels of contrast (1.0% and 0.5%). Largerholes were bored to accept various RMI plugs with known electrondensities for HU accuracy evaluation. The modules can be inserted intoan acrylic tube long enough to cover the entire axial FOV and theirpositions adjusted to desired evaluation points. Conclusion: This phantom allows us to measure the LCR and HU accuracy across theaxial coverage within a single acquisition. These metrics can be usedto characterize the impact different trajectories and reconstructionparameters have on clinically relevant image quality performancemetrics. Funding was provided in part by Varian Medical Systems and NIH R01 Grants Nos. CA158446, CA182264, EB018102, and EB000225. The contents of this poster are solely the responsibility of the authors and do not necessarily represent the official view of any of the supporting organizations.« less

Determination of beam-position dependent transfer functions of LCR-G gravimeters by means of moving mass calibration device in the Mátyáshegy Gravity and Geodynamical Observatory, Budapest

NASA Astrophysics Data System (ADS)

Koppán, András; Kis, Márta; Merényi, László; Papp, Gábor; Benedek, Judit; Meurers, Bruno

2017-04-01

In this presentation authors propose a method for the determination of transfer characteristics and fine calibration of LCR relative gravimeters used for earth-tide recordings, by means of the moving-mass gravimeter calibration device of Budapest-Mátyáshegy Gravity and Geodynamical Observatory. Beam-position dependent transfer functions of four relative LCR G type gravimeters were determined and compared. In order to make these instruments applicable for observatory tidal recordings, there is a need for examining the unique characteristics of equipments and adequately correcting these inherent distorting effects. Thus, the sensitivity for the tilting, temporal changes of scale factors and beam-position dependent transfer characteristics are necessary to be determined for observatory use of these instruments. During the calibration a cylindrical ring of 3200 kg mass is vertically moving around the equipment, generating gravity variations. The effect of the moving mass can be precisely calculated from the known mass and geometrical parameters. The maximum theoretical gravity variation produced by the vertical movement of the mass is ab. 110 microGal, so it provides excellent possibility for the fine calibration of gravimeters in the tidal range. Magnetic experiments were also carried out on the pillar of the calibration device as well, in order to analyse the magnetic effect of the moving stainless steel-mass. According to the magnetic measurements, a correction for the magnetic effect was applied on the measured gravimetric data series. The calibration process is aided by intelligent controller electronics. A PLC-based system has been developed to allow easy control of the movement of the calibrating mass and to measure the mass position. It enables also programmed steps of movements (waiting positions and waiting times) for refined gravity changes. All parameters (position of the mass, CPI data, X/Y leveling positions) are recorded with 1/sec. sampling rate. The system can be controlled remotely through the internet. Authors wish to express their thanks to OTKA (Hungarian Scientific Research Fund) for their support (OTKA-K101603, OTKA K109060).

Unique LCR variations among lineages of HPV16, 18 and 45 isolates from women with normal cervical cytology in Ghana.

PubMed

Awua, Adolf K; Adanu, Richard M K; Wiredu, Edwin K; Afari, Edwin A; Zubuch, Vanessa A; Asmah, Richard H; Severini, Alberto

2017-04-21

In addition to being useful for classification, sequence variations of human Papillomavirus (HPV) genotypes have been implicated in differential oncogenic potential and a differential association with the different histological forms of invasive cervical cancer. These associations have also been indicated for HPV genotype lineages and sub-lineages. In order to better understand the potential implications of lineage variation in the occurrence of cervical cancers in Ghana, we studied the lineages of the three most prevalent HPV genotypes among women with normal cytology as baseline to further studies. Of previously collected self- and health personnel-collected cervical specimen, 54, which were positive for HPV16, 18 and 45, were selected and the long control region (LCR) of each HPV genotype was separately amplified by a nested PCR. DNA sequences of 41 isolates obtained with the forward and reverse primers by Sanger sequencing were analysed. Nucleotide sequence variations of the HPV16 genotypes were observed at 30 positions within the LCR (7460 - 7840). Of these, 19 were the known variations for the lineages B and C (African lineages), while the other 11 positions had variations unique to the HPV16 isolates of this study. For the HPV18 isolates, the variations were at 35 positions, 22 of which were known variations of Africa lineages and the other 13 were unique variations observed for the isolates obtained in this study (at positions 7799 and 7813). HPV45 isolates had variations at 35 positions and 2 (positions 7114 and 97) were unique to the isolates of this study. This study provides the first data on the lineages of HPV 16, 18 and 45 isolates from Ghana. Although the study did not obtain full genome sequence data for a comprehensive comparison with known lineages, these genotypes were predominately of the Africa lineages and had some unique sequence variations at positions that suggest potential oncogenic implications. These data will be useful for comparison with lineages of these genotypes from women with cervical lesion and all the forms of invasive cervical cancers.

Virulence plasmid (pYV)-associated expression of phenotypic virulent determinants in pathogenic Yersinia species: a convenient method for monitoring the presence of pYV under culture conditions and its application for...food

USDA-ARS?s Scientific Manuscript database

In Yersinia pestis, Y. pseudotuberculosis, and Y, enterocolitica, phenotypic expression of virulence plasmid (pYV: 70-kb)-associated genetic determinants may include low calcium response (Lcr, pin point colony, size = 0.36 mm), colony morphology (size = 1.13 mm), crystal violet (CV) binding (dark-v...

Virulence plasmid (pYV)-associated expression of phenotypic virulent determinants in pathogenic Yersinia species: a convenient method for monitoring the presence of pYV under culture conditions and its application for....food

USDA-ARS?s Scientific Manuscript database

In Yersinia pestis, Y. pseudotuberculosis, and Y, enterocolitica, phenotypic expression of several virulence plasmid (pYV: 70-kb)-associated genetic determinants may include low calcium response (Lcr, pin point colony, size = 0.36 mm), colony morphology (size = 1.13 mm), crystal violet (CV) binding...

The locus control region is required for association of the murine β-globin locus with engaged transcription factories during erythroid maturation

PubMed Central

Ragoczy, Tobias; Bender, M.A.; Telling, Agnes; Byron, Rachel; Groudine, Mark

2006-01-01

We have examined the relationship between nuclear localization and transcriptional activity of the endogenous murine β-globin locus during erythroid differentiation. Murine fetal liver cells were separated into distinct erythroid maturation stages by fluorescence-activated cell sorting, and the nuclear position of the locus was determined at each stage. We find that the β-globin locus progressively moves away from the nuclear periphery with increasing maturation. Contrary to the prevailing notion that the nuclear periphery is a repressive compartment in mammalian cells, βmajor-globin expression begins at the nuclear periphery prior to relocalization. However, relocation of the locus to the nuclear interior with maturation is accompanied by an increase in βmajor-globin transcription. The distribution of nuclear polymerase II (Pol II) foci also changes with erythroid differentiation: Transcription factories decrease in number and contract toward the nuclear interior. Moreover, both efficient relocalization of the β-globin locus from the periphery and its association with hyperphosphorylated Pol II transcription factories require the locus control region (LCR). These results suggest that the LCR-dependent association of the β-globin locus with transcriptionally engaged Pol II foci provides the driving force for relocalization of the locus toward the nuclear interior during erythroid maturation. PMID:16705039

A critical analysis of single-frequency LCR databridge impedance measurements of human skin.

PubMed

White, Erick A; Orazem, Mark E; Bunge, Annette L

2011-06-01

Testing whether the barrier of skin samples has sufficient integrity for meaningful measurements of in-vitro chemical permeability is usually required when data are generated for regulatory purposes. Recently, skin integrity has been assessed using LCR databridge measurements, which are reported as resistances determined in either series (SER) or parallel (PAR) modes at a single frequency, typically 100 or 1000Hz. Measurements made at different combinations of mode and frequency are known to differ, although the skin literature reveals confusion over the meaning of these differences and the impact on the interpretation of integrity test results. Here, the theoretical meanings of resistance and capacitance measurements in PAR and SER mode are described and confirmed experimentally. SER-mode resistances are equal to the real part of the complex impedance; whereas, PAR-mode resistances are the inverse of the real part of the admittance. Capacitance measurements reported in SER and PAR modes are similar manipulations of the imaginary parts of the complex impedance and admittance. A large body of data from human cadaver skin is used to show that the PAR-mode resistance and SER-mode capacitance measured at 100Hz are sensitive to skin resistivity, which is the electrical measurement most closely related to skin integrity. Copyright © 2011 Elsevier Ltd. All rights reserved.

Structural polymorphism exhibited by a quasipalindrome present in the locus control region (LCR) of the human beta-globin gene cluster.

PubMed

Kaushik, Mahima; Kukreti, Shrikant

2006-01-01

Structural polymorphism of DNA is a widely accepted property. A simple addition to this perception has been our recent finding, where a single nucleotide polymorphism (SNP) site present in a quasipalindromic sequence of beta-globin LCR exhibited a hairpin-duplex equilibrium. Our current studies explore that secondary structures adopted by individual complementary strands compete with formation of a perfect duplex. Using gel-electrophoresis, ultraviolet (UV)-thermal denaturation, circular dichroism (CD) techniques, we have demonstrated the structural transitions within a perfect duplex containing 11 bp quasipalindromic stretch (TGGGG(G/C)CCCCA), to hairpins and bulge duplex forms. The extended version of the 11 bp duplex, flanked by 5 bp on both sides also demonstrated conformational equilibrium between duplex and hairpin species. Gel-electrophoresis confirms that the duplex coexists with hairpin and bulge duplex/cruciform species. Further, in CD spectra of duplexes, presence of two overlapping positive peaks at 265 and 285 nm suggest the features of A- as well as B-type DNA conformation and show oligomer concentration dependence, manifested in A --> B transition. This indicates the possibility of an architectural switching at quasipalindromic region between linear duplex to a cruciform structure. Such DNA structural variations are likely to be found in the mechanics of molecular recognition and manipulation by proteins.

Rapamycin increases grip strength and attenuates age-related decline in maximal running distance in old low capacity runner rats.

PubMed

Xue, Qian-Li; Yang, Huanle; Li, Hui-Fen; Abadir, Peter M; Burks, Tyesha N; Koch, Lauren G; Britton, Steven L; Carlson, Joshua; Chen, Laura; Walston, Jeremy D; Leng, Sean X

2016-04-01

Rapamycin is known to extend lifespan. We conducted a randomized placebo-controlled study of enteric rapamycin-treatment to evaluate its effect on physical function in old low capacity runner (LCR) rats, a rat model selected from diverse genetic background for low intrinsic aerobic exercise capacity without genomic manipulation and characterized by increased complex disease risks and aging phenotypes. The study was performed in 12 male and 16 female LCR rats aged 16-22 months at baseline. The treatment group was fed with rapamycin-containing diet pellets at approximately 2.24mg/kg body weight per day and the placebo group with the same diet without rapamycin for six months. Observation was extended for additional 2 months. Physical function measurements include grip strength measured as maximum tensile force using a rat grip strength meter and maximum running distance (MRD) using rat physical treadmill test. The results showed that rapamycin improved grip strength by 13% (p=.036) and 60% (p=.001) from its baseline in female and male rats, respectively. Rapamycin attenuated MRD decline by 66% (p=.001) and 46% (p=.319) in females and males, respectively. These findings provide initial evidence for beneficial effect of rapamycin on physical functioning in an aging rat model of high disease risks with significant implication in humans.

Validating Fricke dosimetry for the measurement of absorbed dose to water for HDR 192Ir brachytherapy: a comparison between primary standards of the LCR, Brazil, and the NRC, Canada

NASA Astrophysics Data System (ADS)

Salata, Camila; Gazineu David, Mariano; de Almeida, Carlos Eduardo; El Gamal, Islam; Cojocaru, Claudiu; Mainegra-Hing, Ernesto; McEwen, Malcom

2018-04-01

Two Fricke-based absorbed dose to water standards for HDR Ir-192 dosimetry, developed independently by the LCR in Brazil and the NRC in Canada have been compared. The agreement in the determination of the dose rate from a HDR Ir-192 source at 1 cm in a water phantom was found to be within the k  =  1 combined measurement uncertainties of the two standards: D NRC/D LCR  =  1.011, standard uncertainty  =  2.2%. The dose-based standards also agreed within the uncertainties with the manufacturer’s stated dose rate value, which is traceable to a national standard of air kerma. A number of possible influence quantities were investigated, including the specific method for producing the ferrous-sulphate Fricke solution, the geometry of the holder, and the Monte Carlo code used to determine correction factors. The comparison highlighted the lack of data on the determination of G(Fe3+) in this energy range and the possibilities for further development of the holders used to contain the Fricke solution. The comparison also confirmed the suitability of Fricke dosimetry for Ir-192 primary standard dose rate determinations at therapy dose levels.

Retroviral vectors encoding ADA regulatory locus control region provide enhanced T-cell-specific transgene expression.

PubMed

Trinh, Alice T; Ball, Bret G; Weber, Erin; Gallaher, Timothy K; Gluzman-Poltorak, Zoya; Anderson, French; Basile, Lena A

2009-12-30

Murine retroviral vectors have been used in several hundred gene therapy clinical trials, but have fallen out of favor for a number of reasons. One issue is that gene expression from viral or internal promoters is highly variable and essentially unregulated. Moreover, with retroviral vectors, gene expression is usually silenced over time. Mammalian genes, in contrast, are characterized by highly regulated, precise levels of expression in both a temporal and a cell-specific manner. To ascertain if recapitulation of endogenous adenosine deaminase (ADA) expression can be achieved in a vector construct we created a new series of Moloney murine leukemia virus (MuLV) based retroviral vector that carry human regulatory elements including combinations of the ADA promoter, the ADA locus control region (LCR), ADA introns and human polyadenylation sequences in a self-inactivating vector backbone. A MuLV-based retroviral vector with a self-inactivating (SIN) backbone, the phosphoglycerate kinase promoter (PGK) and the enhanced green fluorescent protein (eGFP), as a reporter gene, was generated. Subsequent vectors were constructed from this basic vector by deletion or addition of certain elements. The added elements that were assessed are the human ADA promoter, human ADA locus control region (LCR), introns 7, 8, and 11 from the human ADA gene, and human growth hormone polyadenylation signal. Retroviral vector particles were produced by transient three-plasmid transfection of 293T cells. Retroviral vectors encoding eGFP were titered by transducing 293A cells, and then the proportion of GFP-positive cells was determined using fluorescence-activated cell sorting (FACS). Non T-cell and T-cell lines were transduced at a multiplicity of infection (MOI) of 0.1 and the yield of eGFP transgene expression was evaluated by FACS analysis using mean fluorescent intensity (MFI) detection. Vectors that contained the ADA LCR were preferentially expressed in T-cell lines. Further improvements in T-cell specific gene expression were observed with the incorporation of additional cis-regulatory elements, such as a human polyadenylation signal and intron 7 from the human ADA gene. These studies suggest that the combination of an authentically regulated ADA gene in a murine retroviral vector, together with additional locus-specific regulatory refinements, will yield a vector with a safer profile and greater efficacy in terms of high-level, therapeutic, regulated gene expression for the treatment of ADA-deficient severe combined immunodeficiency.

Influence of temperature, filter wettability, and timing of filtration on the removal of WBCs from RBC concentrates.

PubMed

van der Meer, P F; Pietersz, R N; Reesink, H W

2001-04-01

The efficacy of the removal of WBCs from buffy coat-reduced RBC concentrates by filtration is determined by many variables. The aim of this study was to investigate the influence of the filtration temperature, the wettability of the filter material, and the timing of the filtration after collection. The investigation used commercially available filters: 3 dry "online" filters (Cellselect FR, Fresenius Hemocare; BioR-01-max, Fresenius; Leucoflex LCG1, MacoPharma) and one wet "inline" filter (Leucoflex LCR4, MacoPharma) that contained saline-adenine-glucose-mannitol additive solution for RBCs and differed from the online version only in wettability. After buffy coat removal and suspension in saline-adenine-glucose-mannitol, filtrations were performed immediately at room temperature (RT) and after 2 hours' storage of the RBC concentrates at 4 degrees C, while the Leucoflex LCR4 was also tested after 24 hours' storage of the RBC concentrates at 4 degrees C. Sets of 12 pooled experiments were performed to prevent donor-dependent differences. The Cellselect FR gave significantly better WBC removal from RBC concentrates at 4 degrees C than at RT, with residual WBCs of 1.44 +/- 0.58 x 10(6) and 2.78 +/- 1.23 x 10(6), respectively (p<0.001). The BioR-01-max gave no significant difference: 0.62 +/- 0.27 x 10(6) WBCs (at 4 degrees C) versus 0.61 +/- 0.25 x 10(6) WBCs (at RT). Filtration with the Leucoflex LCG1 resulted in 0.06 +/- 0.03 x 10(6) and 0.07 +/- 0.07 x 10(6) WBCs at 4 degrees C and RT, respectively, which is not a significant difference. The Leucoflex LCR4, however, gave 2.08 +/- 0.84 x 10(6) WBCs at RT, 0.52 +/- 0.44 x 10(6) WBCs at 4 degrees C after 2 hours' cooling, and 0.05 +/- 0.10 x 10(6) WBCs at 4 degrees C after 24 hours' cooling (all p<0.001). Temperature, filter wettability, and timing of filtration after collection influence the efficacy of a filter for RBC concentrates. These variables need to be established, validated, and controlled before a filter can be selected for routine use.

[Application of Bayes Probability Model in Differentiation of Yin and Yang Jaundice Syndromes in Neonates].

PubMed

Mu, Chun-sun; Zhang, Ping; Kong, Chun-yan; Li, Yang-ning

2015-09-01

To study the application of Bayes probability model in differentiating yin and yang jaundice syndromes in neonates. Totally 107 jaundice neonates who admitted to hospital within 10 days after birth were assigned to two groups according to syndrome differentiation, 68 in the yang jaundice syndrome group and 39 in the yin jaundice syndrome group. Data collected for neonates were factors related to jaundice before, during and after birth. Blood routines, liver and renal functions, and myocardial enzymes were tested on the admission day or the next day. Logistic regression model and Bayes discriminating analysis were used to screen factors important for yin and yang jaundice syndrome differentiation. Finally, Bayes probability model for yin and yang jaundice syndromes was established and assessed. Factors important for yin and yang jaundice syndrome differentiation screened by Logistic regression model and Bayes discriminating analysis included mothers' age, mother with gestational diabetes mellitus (GDM), gestational age, asphyxia, or ABO hemolytic diseases, red blood cell distribution width (RDW-SD), platelet-large cell ratio (P-LCR), serum direct bilirubin (DBIL), alkaline phosphatase (ALP), cholinesterase (CHE). Bayes discriminating analysis was performed by SPSS to obtain Bayes discriminant function coefficient. Bayes discriminant function was established according to discriminant function coefficients. Yang jaundice syndrome: y1= -21. 701 +2. 589 x mother's age + 1. 037 x GDM-17. 175 x asphyxia + 13. 876 x gestational age + 6. 303 x ABO hemolytic disease + 2.116 x RDW-SD + 0. 831 x DBIL + 0. 012 x ALP + 1. 697 x LCR + 0. 001 x CHE; Yin jaundice syndrome: y2= -33. 511 + 2.991 x mother's age + 3.960 x GDM-12. 877 x asphyxia + 11. 848 x gestational age + 1. 820 x ABO hemolytic disease +2. 231 x RDW-SD +0. 999 x DBIL +0. 023 x ALP +1. 916 x LCR +0. 002 x CHE. Bayes discriminant function was hypothesis tested and got Wilks' λ =0. 393 (P =0. 000). So Bayes discriminant function was proved to be with statistical difference. To check Bayes probability model in discriminating yin and yang jaundice syndromes, coincidence rates for yin and yang jaundice syndromes were both 90% plus. Yin and yang jaundice syndromes in neonates could be accurately judged by Bayesian discriminating functions.

Some novel insights on HPV16 related cervical cancer pathogenesis based on analyses of LCR methylation, viral load, E7 and E2/E4 expressions.

PubMed

Das Ghosh, Damayanti; Bhattacharjee, Bornali; Sen, Shrinka; Premi, Laikangbam; Mukhopadhyay, Indranil; Chowdhury, Rahul Roy; Roy, Sudipta; Sengupta, Sharmila

2012-01-01

This study was undertaken to decipher the interdependent roles of (i) methylation within E2 binding site I and II (E2BS-I/II) and replication origin (nt 7862) in the long control region (LCR), (ii) expression of viral oncogene E7, (iii) expression of the transcript (E7-E1/E4) that encodes E2 repressor protein and (iv) viral load, in human papillomavirus 16 (HPV16) related cervical cancer (CaCx) pathogenesis. The results revealed over-representation (p<0.001) of methylation at nucleotide 58 of E2BS-I among E2-intact CaCx cases compared to E2-disrupted cases. Bisulphite sequencing of LCR revealed overrepresentation of methylation at nucleotide 58 or other CpGs in E2BS-I/II, among E2-intact cases than E2-disrupted cases and lack of methylation at replication origin in case of both. The viral transcript (E7-E1/E4) that produces the repressor E2 was analyzed by APOT (amplification of papillomavirus oncogenic transcript)-coupled-quantitative-RT-PCR (of E7 and E4 genes) to distinguish episomal (pure or concomitant with integrated) from purely integrated viral genomes based on the ratio, E7 C(T)/E4 C(T). Relative quantification based on comparative C(T) (threshold cycle) method revealed 75.087 folds higher E7 mRNA expression in episomal cases over purely integrated cases. Viral load and E2 gene copy numbers were negatively correlated with E7 C(T) (p = 0.007) and E2 C(T) (p<0.0001), respectively, each normalized with ACTB C(T), among episomal cases only. The k-means clustering analysis considering E7 C(T) from APOT-coupled-quantitative-RT-PCR assay, in conjunction with viral load, revealed immense heterogeneity among the HPV16 positive CaCx cases portraying integrated viral genomes. The findings provide novel insights into HPV16 related CaCx pathogenesis and highlight that CaCx cases that harbour episomal HPV16 genomes with intact E2 are likely to be distinct biologically, from the purely integrated viral genomes in terms of host genes and/or pathways involved in cervical carcinogenesis.

Chaotic Dynamics and Application of LCR Oscillators Sharing Common Nonlinearity

NASA Astrophysics Data System (ADS)

Jeevarekha, A.; Paul Asir, M.; Philominathan, P.

2016-06-01

This paper addresses the problem of sharing common nonlinearity among nonautonomous and autonomous oscillators. By choosing a suitable common nonlinear element with the driving point characteristics capable of bringing out chaotic motion in a combined system, we obtain identical chaotic states. The dynamics of the coupled system is explored through numerical and experimental studies. Employing the concept of common nonlinearity, a simple chaotic communication system is modeled and its performance is verified through Multisim simulation.

Ontogenesis of endangered humpback chub (Gila cypha) in the Little Colorado River, Arizona

USGS Publications Warehouse

Stone, Dennis M.; Gorman, Owen T.

2006-01-01

The largest population of endangered humpback chub Gila cypha inhabits the Colorado River below Glen Canyon Dam and the lower 14 km of the Little Colorado River (LCR), Arizona. Currently, adults from both rivers spawn and their progenies grow and recruit to adulthood primarily within the LCR, where we studied G. cypha's life history using hoop net capture data. Humpback chub undergo an ontogenesis from diurnally active, vulnerable, nearshore-reliant young-of-the-year (YOY; 30-90 mm total length) into nocturnally active, large-bodied adults (a?Y180 mm TL). During the day, adults primarily resided in deep midchannel pools; however, at night they dispersed inshore amongst the higher densities of YOY conspecifics. Many YOY G. cypha shifted to nocturnal habitats that provided greater cover, possibly, to avoid inshore invading adults. These findings mirror predator-prey scenarios described in other freshwater assemblages, but do not refute other plausible hypotheses. Gila cypha piscivorous activity may escalate in hoop nets, which can confine fish of disparate sizes together; adults were significantly associated with YOY conspecifics and small dead fish in hoop nets at night and eight G. cypha (156-372 mm TL) regurgitated and/or defecated other fish body parts during handling following capture. Gila cypha can definitely be piscivorous given the opportunity, but the magnitude of their piscivorous activity in the wild is debatable.

Changes in productivity and contaminants in bald eagles nesting along the lower Columbia River, USA

USGS Publications Warehouse

Buck, J.A.; Anthony, R.G.; Schuler, C.A.; Isaacs, F.B.; Tillitt, D.E.

2005-01-01

Previous studies documented poor productivity of bald eagles (Haliaeetus leucocephalus) in the lower Columbia River (LCR), USA, and elevated p,p???-dichlorodiphenyldichloroethylene (DDE), polychlorinated biphenyls (PCBs), dioxins, and furans in eagle eggs. From 1994 to 1995, we collected partially incubated eggs at 19 of 43 occupied territories along the LCR and compared productivity and egg contaminants to values obtained in the mid-1980s. We found higher productivity at new nesting sites along the river, yet productivity at 23 older breeding territories remained low and was not different (p = 0.713) between studies. Eggshell thickness at older territories had not improved (p = 0.404), and eggshells averaged 11% thinner than shells measured before dichlorodiphenyltrichloroethane use. Decreases in DDE (p = 0.022) and total PCBs (p = 0.0004) in eggs from older breeding areas occurred between study periods. Productivity was not correlated to contaminants, but DDE, PCBs, and dioxin-like chemicals exceeded estimated no-effect values. Some dioxin-like contaminants in eggs were correlated to nest location, with highest concentrations occurring toward the river's mouth where productivity was lowest. Although total productivity increased due to the success of new nesting pairs in the region, egg contaminants remain high enough to impair reproduction at older territories and, over time, may alter productivity of new pairs nesting near the river's mouth. ?? 2005 SETAC.

Improvement of health risk factors after reduction of VOC concentrations in industrial and urban areas.

PubMed

Lerner, Jorge Esteban Colman; Kohajda, Tibor; Aguilar, Myriam Elisabeth; Massolo, Laura Andrea; Sánchez, Erica Yanina; Porta, Atilio Andrés; Opitz, Philipp; Wichmann, Gunnar; Herbarth, Olf; Mueller, Andrea

2014-01-01

After reductions of fugitive and diffuse emissions by an industrial complex, a follow-up study was performed to determine the time variability of volatile organic compounds (VOCs) and the lifetime cancer risk (LCR). Passive samplers (3 M monitors) were placed outdoors (n = 179) and indoors (n = 75) in industrial, urban, and control areas for 4 weeks. Twenty-five compounds including n-alkanes, cycloalkanes, aromatics, chlorinated hydrocarbons, and terpenes were determined by GC/MS. The results show a significant decrease of all VOCs, especially in the industrial area and to a lesser extent in the urban area. The median outdoor concentration of benzene in the industrial area declined compared to the former study, around 85% and about 50% in the urban area, which in the past was strongly influenced by industrial emissions. Other carcinogenic compounds like styrene and tetrachloroethylene were reduced to approximately 60%. VOC concentrations in control areas remained nearly unchanged. According to the determined BTEX ratios and interspecies correlations, in contrast to the previous study, traffic was identified as the main emission source in the urban and control areas and showed an increased influence in the industrial area. The LCR, calculated for benzene, styrene, and tetrachloroethylene, shows a decrease of one order of magnitude in accordance to the decreased total VOC concentrations and is now acceptable according to values proposed by the World Health Organization.

Rapamycin increases grip strength and attenuates age-related decline in maximal running distance in old low capacity runner rats

PubMed Central

Xue, Qian-Li; Yang, Huanle; Li, Hui-Fen; Abadir, Peter M.; Burks, Tyesha N.; Koch, Lauren G.; Britton, Steven L.; Carlson, Joshua; Chen, Laura; Walston, Jeremy D.; Leng, Sean X.

2016-01-01

Rapamycin is known to extend lifespan. We conducted a randomized placebo-controlled study of enteric rapamycin-treatment to evaluate its effect on physical function in old low capacity runner (LCR) rats, a rat model selected from diverse genetic background for low intrinsic aerobic exercise capacity without genomic manipulation and characterized by increased complex disease risks and aging phenotypes. The study was performed in 12 male and 16 female LCR rats aged 16-22 months at baseline. The treatment group was fed with rapamycin-containing diet pellets at approximately 2.24mg/kg body weight per day and the placebo group with the same diet without rapamycin for six months. Observation was extended for additional 2 months. Physical function measurements include grip strength measured as maximum tensile force using a rat grip strength meter and maximum running distance (MRD) using rat physical treadmill test. The results showed that rapamycin improved grip strength by 13% (p=.036) and 60% (p<.001) from its baseline in female and male rats, respectively. Rapamycin attenuated MRD decline by 66% (p<.001) and 46% (p=.319) in females and males, respectively. These findings provide initial evidence for beneficial effect of rapamycin on physical functioning in an aging rat model of high disease risks with significant implication in humans. PMID:26997106

Label-Free LC-MS Profiling of Skeletal Muscle Reveals Heart-Type Fatty Acid Binding Protein as a Candidate Biomarker of Aerobic Capacity.

PubMed

Malik, Zulezwan Ab; Cobley, James N; Morton, James P; Close, Graeme L; Edwards, Ben J; Koch, Lauren G; Britton, Steven L; Burniston, Jatin G

2013-12-01

Two-dimensional gel electrophoresis provides robust comparative analysis of skeletal muscle, but this technique is laborious and limited by its inability to resolve all proteins. In contrast, orthogonal separation by SDS-PAGE and reverse-phase liquid chromatography (RPLC) coupled to mass spectrometry (MS) affords deep mining of the muscle proteome, but differential analysis between samples is challenging due to the greater level of fractionation and the complexities of quantifying proteins based on the abundances of their tryptic peptides. Here we report simple, semi-automated and time efficient ( i.e ., 3 h per sample) proteome profiling of skeletal muscle by 1-dimensional RPLC electrospray ionisation tandem MS. Solei were analysed from rats (n = 5, in each group) bred as either high- or low-capacity runners (HCR and LCR, respectively) that exhibited a 6.4-fold difference (1,625 ± 112 m vs . 252 ± 43 m, p < 0.0001) in running capacity during a standardized treadmill test. Soluble muscle proteins were extracted, digested with trypsin and individual biological replicates (50 ng of tryptic peptides) subjected to LC-MS profiling. Proteins were identified by triplicate LC-MS/MS analysis of a pooled sample of each biological replicate. Differential expression profiling was performed on relative abundances (RA) of parent ions, which spanned three orders of magnitude. In total, 207 proteins were analysed, which encompassed almost all enzymes of the major metabolic pathways in skeletal muscle. The most abundant protein detected was type I myosin heavy chain (RA = 5,843 ± 897) and the least abundant protein detected was heat shock 70 kDa protein (RA = 2 ± 0.5). Sixteen proteins were significantly ( p < 0.05) more abundant in HCR muscle and hierarchal clustering of the profiling data highlighted two protein subgroups, which encompassed proteins associated with either the respiratory chain or fatty acid oxidation. Heart-type fatty acid binding protein (FABPH) was 1.54-fold ( p = 0.0064) more abundant in HCR than LCR soleus. This discovery was verified using selective reaction monitoring (SRM) of the y5 ion (551.21 m/z ) of the doubly-charged peptide SLGVGFATR (454.19 m/z ) of residues 23-31 of FABPH. SRM was conducted on technical replicates of each biological sample and exhibited a coefficient of variation of 20%. The abundance of FABPH measured by SRM was 2.84-fold greater ( p = 0.0095) in HCR muscle. In addition, SRM of FABPH was performed in vastus lateralis samples of young and elderly humans with different habitual activity levels (collected during a previous study) finding FABPH abundance was 2.23-fold greater ( p = 0.0396) in endurance-trained individuals regardless of differences in age. In summary, our findings in HCR/LCR rats provide protein-level confirmation for earlier transcriptome profiling work and show LC-MS is a viable means of profiling the abundance of almost all major metabolic enzymes of skeletal muscle in a highly parallel manner. Moreover, our approach is relatively more time efficient than techniques relying on orthogonal separations, and we demonstrate LC-MS profiling of the HCR/LCR selection model was able to highlight biomarkers that also exhibit differences in trained and untrained human muscle.

An Epidemiologic Analysis of Chlamydia trachomatis and Neisseria gonorrhoeae Infections in Female Federal Prisoners

DTIC Science & Technology

2002-01-01

compared to culture.70,72 Use of PCR to detect GC has yielded sensitivities between 92-100% and specificities ranging from 96-99% on endocervical...specimen.73,74 With use of urine specimens, PCR has yielded sensitivities ranging from 65 to 92% and 96-99% respectively, for GC detection .74...21 An important limitation of PCR and LCR is that the tests may detect CT or GC nucleic acid remaining after therapy has been administered

Intrinsic aerobic capacity correlates with greater inherent mitochondrial oxidative and H2O2 emission capacities without major shifts in myosin heavy chain isoform.

PubMed

Seifert, Erin L; Bastianelli, Mark; Aguer, Céline; Moffat, Cynthia; Estey, Carmen; Koch, Lauren G; Britton, Steven L; Harper, Mary-Ellen

2012-11-01

Exercise capacity and performance strongly associate with metabolic and biophysical characteristics of skeletal muscle, factors that also relate to overall disease risk. Despite its importance, the exact mechanistic features that connect aerobic metabolism with health status are unknown. To explore this, we applied artificial selection of rats for intrinsic (i.e., untrained) aerobic treadmill running to generate strains of low- and high-capacity runners (LCR and HCR, respectively), subsequently shown to diverge for disease risk. Concurrent breeding of LCR and HCR per generation allows the lines to serve as reciprocal controls for unknown environmental changes. Here we provide the first direct evidence in mitochondria isolated from skeletal muscle that intrinsic mitochondrial capacity is higher in HCR rats. Maximal phosphorylating respiration was ~40% greater in HCR mitochondria, independent of substrate and without altered proton leak or major changes in protein levels or muscle fiber type, consistent with altered control of phosphorylating respiration. Unexpectedly, H(2)O(2) emission was ~20% higher in HCR mitochondria, due to greater reduction of more harmful reactive oxygen species to H(2)O(2); indeed, oxidative modification of mitochondrial proteins was lower. When the higher mitochondrial yield was considered, phosphorylating respiration and H(2)O(2) emission were 70-80% greater in HCR muscle. Greater capacity of HCR muscle for work and H(2)O(2) signaling may result in enhanced and more immediate cellular repair, possibly explaining lowered disease risks.

Yersinia pestis with regulated delayed attenuation as a vaccine candidate to induce protective immunity against plague.

PubMed

Sun, Wei; Roland, Kenneth L; Kuang, Xiaoying; Branger, Christine G; Curtiss, Roy

2010-03-01

Two mutant strains of Yersinia pestis KIM5+, a Deltacrp mutant and a mutant with arabinose-dependent regulated delayed-shutoff crp expression (araC P(BAD) crp), were constructed, characterized in vitro, and evaluated for virulence, immunogenicity, and protective efficacy in mice. Both strains were highly attenuated by the subcutaneous (s.c.) route. The 50% lethal doses (LD(50)s) of the Deltacrp and araC P(BAD) crp mutants were approximately 1,000,000-fold and 10,000-fold higher than those of Y. pestis KIM5+, respectively, indicating that both strains were highly attenuated. Mice vaccinated s.c. with 3.8 x 10(7) CFU of the Deltacrp mutant developed high anti-Y. pestis and anti-LcrV serum IgG titers, both with a strong Th2 bias, and induced protective immunity against subcutaneous challenge with virulent Y. pestis (80% survival) but no protection against pulmonary challenge. Mice vaccinated with 3.0 x 10(4) CFU of the araC P(BAD) crp mutant also developed high anti-Y. pestis and anti-LcrV serum IgG titers but with a more balanced Th1/Th2 response. This strain induced complete protection against s.c. challenge and partial protection (70% survival) against pulmonary challenge. Our results demonstrate that arabinose-dependent regulated crp expression is an effective strategy to attenuate Y. pestis while retaining strong immunogenicity, leading to protection against the pneumonic and bubonic forms of plague.

Electroporation of a multivalent DNA vaccine cocktail elicits a protective immune response against anthrax and plague.

PubMed

Albrecht, Mark T; Livingston, Brian D; Pesce, John T; Bell, Matt G; Hannaman, Drew; Keane-Myers, Andrea M

2012-07-06

Electroporation of DNA vaccines represents a platform technology well positioned for the development of multivalent biodefense vaccines. To evaluate this hypothesis, three vaccine constructs were produced using codon-optimized genes encoding Bacillus anthracis Protective Antigen (PA), and the Yersinia pestis genes LcrV and F1, cloned into pVAX1. A/J mice were immunized on a prime-boost schedule with these constructs using the electroporation-based TriGrid Delivery System. Immunization with the individual pDNA vaccines elicited higher levels of antigen-specific IgG than when used in combination. DNA vaccine effectiveness was proven, the pVAX-PA titers were toxin neutralizing and fully protective against a lethal B. anthracis spore challenge when administered alone or co-formulated with the plague pDNA vaccines. LcrV and F1 pVAX vaccines against plague were synergistic, resulting in 100% survival, but less protective individually and when co-formulated with pVAX-PA. These DNA vaccine responses were Th1/Th2 balanced with high levels of IFN-γ and IL-4 in splenocyte recall assays, contrary to complimentary protein Alum vaccinations displaying a Th2 bias with increased IL-4 and low levels of IFN-γ. These results demonstrate the feasibility of electroporation to deliver and maintain the overall efficacy of an anthrax-plague DNA vaccine cocktail whose individual components have qualitative immunological differences when combined. Published by Elsevier Ltd.

Synthesis and characterization of barium hexaferrite with manganese (Mn) doping material as anti-radar

NASA Astrophysics Data System (ADS)

Susilawati, Doyan, Aris; Khalilurrahman

2017-01-01

Have been successfully synthesized barium powder doping Manganese hexaferrite with the expected potential as anti-radar material. Synthesis was done by using the co-precipitation method, the variation of the variable x concentrations used were 0; 0.2; 0.4; and 0.6 and calcined at temperatures of 400, 600 and 800°C. Characterization powders of hexaferrite have used XRD (X-Ray Diffraction), SEM (Scanning Electron Microscopy), TEM (Transmission Electron Microscopy), LCR (inductance, capacitance, and resistance) meter, and VSM (Vibrating Sample Magnetometer). The higher the concentration and temperature of calcinations given affect the color of the powder. The test results using XRD indicates that it has formed barium hexaferrite phase with a hexagonal crystal structure. Tests using SEM showed that all the constituent elements barium powder hexaferrite by doping Manganese powders have been spread evenly. XRD test results were confirmed by a test using a TEM showing the crystal structure and the powder was sized nano particles. The results from the LCR meter showed that the barium powder hexaferrite by doping Manganese that has been synthesized classified in semiconductor materials. The result from VSM showed that the value of coercivity magnetic powder doped barium hexaferrite Manganese is smaller when compared with barium hexaferrite without doping and belong to the soft magnetic. Based on the results of the synthesis and characterization, we can conclude that the barium powder heksaferrite by doping Manganese potential as a material anti-radar.

β-Globin locus control region HS2 and HS3 interact structurally and functionally

PubMed Central

Jackson, David A.; McDowell, Jennifer C.; Dean, Ann

2003-01-01

The overall structure of the DNase I hypersensitive sites (HSs) that comprise the β-globin locus control region (LCR) is highly conserved among mammals, implying that the HSs have conserved functions. However, it is not well understood how the LCR HSs, either individually or collectively, activate transcription. We analyzed the interactions of HS2, HS3 and HS4 with the human ε- and β-globin genes in chromatinized episomes in fetal/embryonic K562 cells. Only HS2 activates transcription of the ε-globin gene, while all three HSs activate the β-globin gene. HS3 stimulates the β-globin gene constitutively, but HS2 and HS4 transactivation requires expression of the transcription factor EKLF, which is not present in K562 cells but is required for β-globin expression in vivo. To begin addressing how the individual HSs may interact with one another in a complex, we linked the β-globin gene to both the HS2 and HS3. HS2 and HS3 together resulted in synergistic stimulation of β-globin transcription. Unexpectedly, mutated, inactive forms of HS2 impeded the activation of the β-globin gene by HS3. Thus, there appear to be distinct interactions among the HSs and between the HSs and the globin genes. These preferential, non-exclusive interactions may underlie an important structural and functional cooperativity among the regulatory sequences of the β-globin locus in vivo. PMID:12582237

Synthesis and characterization of PVA blended LiClO4 as electrolyte material for battery Li-ion

NASA Astrophysics Data System (ADS)

Gunawan, I.; Deswita; Sugeng, B.; Sudaryanto

2017-07-01

It have been synthesized the materials for Li ion battery electrolytes, namely PVA with the addition of LiClO4 salt were varied 0, 5, 10, 15 and 20% by weight respectively. The objective of this study is to control the ionic conductivity in traditional polymer electrolytes, to improve ionic conductivity with the addition of lithium perchlorat (LiClO4). These electrolyte materials prepared by PVA powder was dissolved into distilled water and added LiClO4 salt were varied. After drying the solution, PVA sheet blended LiClO4 salt as electrolyte material for Li ion battery obtained. PVA blended LiClO4 salt crystallite form was confirmed using X-Ray Difraction (XRD) equipment. Observation of the morphology done by using Scanning Electron Microscope (SEM). While the electrical conductivity of the material is measured using LCR meter. The results of XRD pattern of LiClO4 shows intense peaks at angles 2θ = 23.2, 32.99, and 36.58°, which represent the crystalline nature of the salt. Particles morphology of the sample revealed by scanning electron microscopy are irregular in shape and agglomerated, with mean size 200-300 nm. It can be concluded that polycrystalline particles are composed of large number of crystallites. The study of conductivity by using LCR meter shows that all the graphs represent the DC and AC conductivity phenomena.

Heavy metal exposure from cooked rice grain ingestion and its potential health risks to humans from total and bioavailable forms analysis.

PubMed

Praveena, S M; Omar, N A

2017-11-15

Heavy metal in rice studies has attracted a greater concern worldwide. However, there have been limited studies on marketed rice samples although it represents a vital ingestion portion for a real estimation of human health risk. This study was aimed to determine both total and bioaccessible of trace elements and heavy metals (Cd, Cr, Cu, Co, Al, Zn, As, Pb and Fe) in 22 varieties of cooked rice using an inductively coupled plasma-optical emission spectroscopy. Both total and bioaccessible of trace elements and heavy metals were digested using closed-nitric acid digestion and Rijksinstituut voor Volksgezondheid en Milieu (RIVM) in vitro digestion model, respectively. Human health risks via Health Risk Assessment (HRA) were conducted to understand exposure risks involving adults and children representing Malaysian population. Zinc was the highest while As was the lowest contents for total and in their bioavailable forms. Four clusters were identified: (1) Pb, As, Co, Cd and Cr; (2) Cu and Al; (3) Fe and (4) Zn. For HRA, there was no any risks found from single element exposure. While potential carcinogenic health risks present for both adult and children from single As exposure (Life time Cancer Risk, LCR>1×10 -4 ). Total Hazard Quotient values for adult and children were 27.0 and 18.0, respectively while total LCR values for adult and children were 0.0049 and 0.0032, respectively. Copyright © 2017 Elsevier Ltd. All rights reserved.

Size distributions of polycyclic aromatic hydrocarbons in urban atmosphere: sorption mechanism and source contributions to respiratory deposition

NASA Astrophysics Data System (ADS)

Lv, Yan; Li, Xiang; Xu, Ting Ting; Cheng, Tian Tao; Yang, Xin; Chen, Jian Min; Iinuma, Yoshiteru; Herrmann, Hartmut

2016-03-01

In order to better understand the particle size distribution of polycyclic aromatic hydrocarbons (PAHs) and their source contribution to human respiratory system, size-resolved PAHs have been studied in ambient aerosols at a megacity Shanghai site during a 1-year period (2012-2013). The results showed the PAHs had a bimodal distribution with one mode peak in the fine-particle size range (0.4-2.1 µm) and another mode peak in the coarse-particle size range (3.3-9.0 µm). Along with the increase in ring number of PAHs, the intensity of the fine-mode peak increased, while the coarse-mode peak decreased. Plotting of log(PAH / PM) against log(Dp) showed that all slope values were above -1, suggesting that multiple mechanisms (adsorption and absorption) controlled the particle size distribution of PAHs. The total deposition flux of PAHs in the respiratory tract was calculated as being 8.8 ± 2.0 ng h-1. The highest lifetime cancer risk (LCR) was estimated at 1.5 × 10-6, which exceeded the unit risk of 10-6. The LCR values presented here were mainly influenced by accumulation mode PAHs which came from biomass burning (24 %), coal combustion (25 %), and vehicular emission (27 %). The present study provides us with a mechanistic understanding of the particle size distribution of PAHs and their transport in the human respiratory system, which can help develop better source control strategies.

SU-E-P-18: Intensity-Modulated Radiation Therapy for Cervical Esophageal Squamous Cell Carcinoma

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

Bai, W; Qiao, X; Zhou, Z

2015-06-15

Purpose: To retrospectively analyze the outcomes and prognostic factors of cervical esophageal squamous cell carcinoma (SCC) treated with intensity modulated radiation therapy (IMRT). Methods: Thirty-seven patients with cervical esophageal SCC treated with IMRT were analyzed retrospectively. They received 54–66 Gy in 27–32 fractions. Nineteen patients received concurrent (n=12) or sequential (n=7) platinum-based two drugs chemoradiotherapy. Overall survival (OS), local control rates (LCR) and prognostic factors were evaluated. Acute toxicities and patterns of first failures were observed. Results: The median follow-up was 46 months for alive patients. The l-, 3-, 4- and 5-year OS of the all patients were 83.8%, 59.1%,more » 47.5% and 32.6% respectively. The median survival time was 46 months. The l-, 3-,4- and 5-year LCR were 82.9%, 63.0%, 54.5% and 54.5%, respectively. Univariate and Multivariate analysis all showed that size of GTV was an independent prognostic factor (p=0.033, p=0.039). There were no patients with Grade 3 acute radiation esophagitis and Grade 2–4 acute pneumonitis. The local failure accounted for 70.0% of all treatment-related failures. Conclusion: IMRT is safe and effective in the treatment of cervical esophageal squamous cell carcinoma. Size of GTV is an independent prognostic factor. Local failure still remains the main reason of treatment failures. The authors declare no conflicts of interest in preparing this article.« less

Retroviral vectors encoding ADA regulatory locus control region provide enhanced T-cell-specific transgene expression

PubMed Central

2009-01-01

Background Murine retroviral vectors have been used in several hundred gene therapy clinical trials, but have fallen out of favor for a number of reasons. One issue is that gene expression from viral or internal promoters is highly variable and essentially unregulated. Moreover, with retroviral vectors, gene expression is usually silenced over time. Mammalian genes, in contrast, are characterized by highly regulated, precise levels of expression in both a temporal and a cell-specific manner. To ascertain if recapitulation of endogenous adenosine deaminase (ADA) expression can be achieved in a vector construct we created a new series of Moloney murine leukemia virus (MuLV) based retroviral vector that carry human regulatory elements including combinations of the ADA promoter, the ADA locus control region (LCR), ADA introns and human polyadenylation sequences in a self-inactivating vector backbone. Methods A MuLV-based retroviral vector with a self-inactivating (SIN) backbone, the phosphoglycerate kinase promoter (PGK) and the enhanced green fluorescent protein (eGFP), as a reporter gene, was generated. Subsequent vectors were constructed from this basic vector by deletion or addition of certain elements. The added elements that were assessed are the human ADA promoter, human ADA locus control region (LCR), introns 7, 8, and 11 from the human ADA gene, and human growth hormone polyadenylation signal. Retroviral vector particles were produced by transient three-plasmid transfection of 293T cells. Retroviral vectors encoding eGFP were titered by transducing 293A cells, and then the proportion of GFP-positive cells was determined using fluorescence-activated cell sorting (FACS). Non T-cell and T-cell lines were transduced at a multiplicity of infection (MOI) of 0.1 and the yield of eGFP transgene expression was evaluated by FACS analysis using mean fluorescent intensity (MFI) detection. Results Vectors that contained the ADA LCR were preferentially expressed in T-cell lines. Further improvements in T-cell specific gene expression were observed with the incorporation of additional cis-regulatory elements, such as a human polyadenylation signal and intron 7 from the human ADA gene. Conclusion These studies suggest that the combination of an authentically regulated ADA gene in a murine retroviral vector, together with additional locus-specific regulatory refinements, will yield a vector with a safer profile and greater efficacy in terms of high-level, therapeutic, regulated gene expression for the treatment of ADA-deficient severe combined immunodeficiency. PMID:20042112

Longitudinal Characteristics and Aileron Effectiveness of a Midwing Airplane from High-Speed Wind-Tunnel Tests

DTIC Science & Technology

1944-09-01

undesirable control cho.r~cteristics caused by the decrease in the elcvator- f’Lxcd pitehinc-moi’aent coefficient (fiZ. 12.(8.)). The effect of the t~b on...LONGITUDINAL CHARACTERISTICS AND AILERON EFFECTIVENESS OF A MIDWING AIRPLANE FROM ffiGH-SPEED WIND-TUNNEL TESTS By Charles F. Hall and Robert L. Mannes Ames...thG o.i Lcr-on effectiveness a t bieh Hc.ch numbers. Tho forco and Gamont coefficients computod fro~ the tost data nrc prcsontcd in this ro)ort. The

Broad T Cell Immunity to the LcrV Virulence Protein is Induced by Targeted Delivery to DEC-205/CD205-Positive Mouse Dendritic Cells

DTIC Science & Technology

2007-08-13

to its potential use as a biological weapon [1, 2]. Bubonic plague is the most common clinical form of disease and is characterized by swollen lymph...naive mice. Prior plague vaccines, such as killed whole-cell vaccine and attenuated vaccine, induce protection against bubonic plague , but the...R. W. and Williamson, E. D., Vaccination against bubonic and pneumonic plague . Vaccine 2001. 19: 4175–4184. 5 Du, Y., Rosqvist, R. and Forsberg, A

Structural and dielectric properties of CTAB modified ZrO2 nanoparticles

NASA Astrophysics Data System (ADS)

Sidhu, Gaganpreet Kaur; Tripathi, S. K.; Kumar, Rajesh

2016-05-01

Zirconia (ZrO2) has been considered as one of the most investigated materials among various metal oxides due its outstanding dielectric properties and ionic conduction properties, which is mainly due to its high oxygen ion conduction. ZrO2 nanoparticles were synthesized using surfactant (CTAB) to study the variation of its dielectric behavior at room temperature. Surfactants form a unique class of chemical compounds, because of their remarkable ability to influence the properties of surfaces and interfaces of nanostructures. The dielectric properties of prepared nanoparticles were studied using LCR meter.

Bibliography on Cold Regions Science and Technology. Volume 36, Part 1, 1982.

DTIC Science & Technology

1982-09-01

western MeNlurdo Sound. and the fact that thewe areas are truly rainless, many features Barrett, P I, ct al. .- lcr st fi p h ilc l I ,i,, . iss i and...beneath structures. Engi- 1981. Nir Mr. P 42 IlC Ressea equipment. ge rittund- leeriim utlmcog. Ertiten rick temperature. Snon Riesal’. A, \\. depth...i Jarri ii soc ctr im treg itjan ra irsd~rI-J River basins, Snot. cover distribution, Soon . water ik. \\iii s icrhr airIr i ii ’r~ ir~c rrrkrr .I

Synthesis and Characterization Materials M-Barium Hexaferrite Doping Ions Co-Mn Nano Particle

NASA Astrophysics Data System (ADS)

Susilawati; Doyan, A.; Sahlam

2017-05-01

This research has been success in the synthesis of M-Barium hexaferrite (BaM) doping Co-Mn ions using coprecipitation method are expected to be applied as a base material in the coating RADAR. M-Barium hexaferrite (BaM) are BaFe12-2xCoxMnxO19 synthesized with various concentrations (x = 0.0, 0.1, 0.2, 0.3) and the calcinations temperature (T = 400°C, 600°C, 800°C). The materials characterization using a X-Ray Diffraction (XRD), Transmission Electron Microscope (TEM), Inductance Capacitance and resistance (LCR) meter, and Vibrating Sample Magnetometer (VSM) Instruments. The measurement results using XRD shows the material has a hexagonal crystalline structure. The measurement results using a TEM show a sample of nano crystal materials with grain diameters up to 40 nm and spacing of the crystal lattice. The measurement results using a LCR-meter shows the electric conductivity of 1.15 × 10-6 S/cm to BaM without doping, 3.75 × 10-6 S/cm to 0.1 doping concentration calcination temperature of 400 °C, and 1,23 × 10-5 S/cm to 0.3 doping concentration calcination temperature of 800 °C, thus including semiconductor materials. The magnetic properties of materials using a VSM test results show the value of coercivity of 0.1 T; remanence value of 0.06 emu/g; and the saturation value of 0.42 emu/g. The results above show BaM Co-Mn metal doping potential as anti-radar material.

Eos Negatively Regulates Human γ-globin Gene Transcription during Erythroid Differentiation

PubMed Central

Yu, Hai-Chuan; Zhao, Hua-Lu; Wu, Zhi-Kui; Zhang, Jun-Wu

2011-01-01

Background Human globin gene expression is precisely regulated by a complicated network of transcription factors and chromatin modifying activities during development and erythropoiesis. Eos (Ikaros family zinc finger 4, IKZF4), a member of the zinc finger transcription factor Ikaros family, plays a pivotal role as a repressor of gene expression. The aim of this study was to examine the role of Eos in globin gene regulation. Methodology/Principal Findings Western blot and quantitative real-time PCR detected a gradual decrease in Eos expression during erythroid differentiation of hemin-induced K562 cells and Epo-induced CD34+ hematopoietic stem/progenitor cells (HPCs). DNA transfection and lentivirus-mediated gene transfer demonstrated that the enforced expression of Eos significantly represses the expression of γ-globin, but not other globin genes, in K562 cells and CD34+ HPCs. Consistent with a direct role of Eos in globin gene regulation, chromatin immunoprecipitaion and dual-luciferase reporter assays identified three discrete sites located in the DNase I hypersensitivity site 3 (HS3) of the β-globin locus control region (LCR), the promoter regions of the Gγ- and Aγ- globin genes, as functional binding sites of Eos protein. A chromosome conformation capture (3C) assay indicated that Eos may repress the interaction between the LCR and the γ-globin gene promoter. In addition, erythroid differentiation was inhibited by enforced expression of Eos in K562 cells and CD34+ HPCs. Conclusions/Significance Our results demonstrate that Eos plays an important role in the transcriptional regulation of the γ-globin gene during erythroid differentiation. PMID:21829552

Intrinsic aerobic capacity correlates with greater inherent mitochondrial oxidative and H2O2 emission capacities without major shifts in myosin heavy chain isoform

PubMed Central

Seifert, Erin L.; Bastianelli, Mark; Aguer, Céline; Moffat, Cynthia; Estey, Carmen; Koch, Lauren G.; Britton, Steven L.

2012-01-01

Exercise capacity and performance strongly associate with metabolic and biophysical characteristics of skeletal muscle, factors that also relate to overall disease risk. Despite its importance, the exact mechanistic features that connect aerobic metabolism with health status are unknown. To explore this, we applied artificial selection of rats for intrinsic (i.e., untrained) aerobic treadmill running to generate strains of low- and high-capacity runners (LCR and HCR, respectively), subsequently shown to diverge for disease risk. Concurrent breeding of LCR and HCR per generation allows the lines to serve as reciprocal controls for unknown environmental changes. Here we provide the first direct evidence in mitochondria isolated from skeletal muscle that intrinsic mitochondrial capacity is higher in HCR rats. Maximal phosphorylating respiration was ∼40% greater in HCR mitochondria, independent of substrate and without altered proton leak or major changes in protein levels or muscle fiber type, consistent with altered control of phosphorylating respiration. Unexpectedly, H2O2 emission was ∼20% higher in HCR mitochondria, due to greater reduction of more harmful reactive oxygen species to H2O2; indeed, oxidative modification of mitochondrial proteins was lower. When the higher mitochondrial yield was considered, phosphorylating respiration and H2O2 emission were 70–80% greater in HCR muscle. Greater capacity of HCR muscle for work and H2O2 signaling may result in enhanced and more immediate cellular repair, possibly explaining lowered disease risks. PMID:22995392

The Virulence Plasmid of Yersinia, an Antihost Genome

PubMed Central

Cornelis, Guy R.; Boland, Anne; Boyd, Aoife P.; Geuijen, Cecile; Iriarte, Maite; Neyt, Cécile; Sory, Marie-Paule; Stainier, Isabelle

1998-01-01

The 70-kb virulence plasmid enables Yersinia spp. (Yersinia pestis, Y. pseudotuberculosis, and Y. enterocolitica) to survive and multiply in the lymphoid tissues of their host. It encodes the Yop virulon, an integrated system allowing extracellular bacteria to disarm the cells involved in the immune response, to disrupt their communications, or even to induce their apoptosis by the injection of bacterial effector proteins. This system consists of the Yop proteins and their dedicated type III secretion apparatus, called Ysc. The Ysc apparatus is composed of some 25 proteins including a secretin. Most of the Yops fall into two groups. Some of them are the intracellular effectors (YopE, YopH, YpkA/YopO, YopP/YopJ, YopM, and YopT), while the others (YopB, YopD, and LcrV) form the translocation apparatus that is deployed at the bacterial surface to deliver the effectors into the eukaryotic cells, across their plasma membrane. Yop secretion is triggered by contact with eukaryotic cells and controlled by proteins of the virulon including YopN, TyeA, and LcrG, which are thought to form a plug complex closing the bacterial secretion channel. The proper operation of the system also requires small individual chaperones, called the Syc proteins, in the bacterial cytosol. Transcription of the genes is controlled both by temperature and by the activity of the secretion apparatus. The virulence plasmid of Y. enterocolitica and Y. pseudotuberculosis also encodes the adhesin YadA. The virulence plasmid contains some evolutionary remnants including, in Y. enterocolitica, an operon encoding resistance to arsenic compounds. PMID:9841674

Human papillomavirus type-16 variants in Quechua aboriginals from Argentina.

PubMed

Picconi, María Alejandra; Alonio, Lidia Virginia; Sichero, Laura; Mbayed, Viviana; Villa, Luisa Lina; Gronda, Jorge; Campos, Rodolfo; Teyssié, Angélica

2003-04-01

Cervical carcinoma is the leading cause of cancer death in Quechua indians from Jujuy (northwestern Argentina). To determine the prevalence of HPV-16 variants, 106 HPV-16 positive cervical samples were studied, including 33 low-grade squamous intraepithelial lesions (LSIL), 28 high-grade squamous intraepithelial lesions (HSIL), 9 invasive cervical cancer (ICC), and 36 samples from women with normal colposcopy and cytology. HPV genome variability was examined in the L1 and E6 genes by PCR-hybridization. In a subset of 20 samples, a LCR fragment was also analyzed by PCR-sequencing. Most variants belonged to the European branch with subtle differences that depended on the viral gene fragment studied. Only about 10% of the specimens had non-European variants, including eight Asian-American, two Asian, and one North-American-1. E6 gene analysis revealed that 43% of the samples were identical to HPV-16 prototype, while 57% corresponded to variants. Interestingly, the majority (87%) of normal smears had HPV-16 prototype, whereas variants were detected mainly in SIL and ICC. LCR sequencing yielded 80% of variants, including 69% of European, 19% Asian-American, and 12% Asian. We identified a new variant, the Argentine Quechua-51 (AQ-51), similar to B-14 plus two additional changes: G7842-->A and A7837-->C; phylogenetic inference allocated it in the Asian-American branch. The high proportion of European variants may reflect Spanish colonial influence on these native Inca descendants. The predominance of HPV-16 variants in pathologic samples when compared to normal controls could have implications for the natural history of cervical lesions. Copyright 2003 Wiley-Liss, Inc.

Electromagnetic induction imaging of concealed metallic objects by means of resonating circuits

NASA Astrophysics Data System (ADS)

Guilizzoni, R.; Watson, J. C.; Bartlett, P. A.; Renzoni, F.

2016-05-01

An electromagnetic induction system, suitable for 2D imaging of metallic samples of different electrical conductivities, has been developed. The system is based on a parallel LCR circuit comprising a ferrite-cored coil (7.8 mm x 9.5 mm, L=680 μH at 1 KHz), a variable resistor and capacitor. The working principle of the system is based on eddy current induction inside a metallic sample when this is introduced into the AC magnetic field created by the coil. The inductance of the LCR circuit is modified due to the presence of the sample, to an extent that depends on its conductivity. Such modification is known to increase when the system is operated at its resonant frequency. Characterizing different metals based on their values of conductivity is therefore possible by utilizing a suitable system operated at resonance. Both imaging and material characterization were demonstrated by means of the proposed electromagnetic induction technique. Furthermore, the choice of using a system with an adjustable resonant frequency made it possible to select resonances that allow magnetic-field penetration through conductive screens. Investigations on the possibility of imaging concealed metals by penetrating such shields have been carried out. A penetration depth of δ~3 mm through aluminium (Al) was achieved. This allowed concealed metallic samples- having conductivities ranging from 0.54 to 59.77 MSm-1 and hidden behind 1.5-mm-thick Al shields- to be imaged. Our results demonstrate that the presence of the concealed metallic objects can be revealed. The technique was thus shown to be a promising detection tool for security applications.

Influence of magnification and superimposition of structures on cephalometric diagnosis.

PubMed

Paula, Leonardo Koerich de; Solon-de-Mello, Priscilla de Almeida; Mattos, Claudia Trindade; Ruellas, Antônio Carlos de Oliveira; Sant'Anna, Eduardo Franzotti

2015-01-01

The purpose of this study was to assess the influence of magnification and superimposition of structures on CBCT-generated lateral cephalometric radiographs (LCR) using different segments of the cranium. CBCT scans of 10 patients were selected. Four LCR were generated using Dolphin Imaging(r) software: full-face, right side, left side and center of the head. A total of 40 images were imported into Radiocef Studio 2(r), and the angles of the most common cephalometric analyses were traced by the same observer twice and within a 10-day interval. Statistical analyses included intraexaminer agreement and comparison between methods by means of intraclass correlation coefficient (ICC) and Bland-Altman agreement tests. Intraexaminer agreement of the angles assessed by ICC was excellent (> 0.90) for 83% of measurements, good (between 0.75 and 0.90) for 15%, and moderate (between 0.50 and 0.75) for 2% of measurements. The comparison between methods by ICC was excellent for 68% of measurements, good for 26%, and moderate for 6%. Variables presenting wider confidence intervals (> 6o) in the Bland-Altman tests, in intraexaminer assessment, were: mandibular incisor angle, maxillary incisor angle, and occlusal plane angle. And in comparison methods the variables with wider confidence interval were: mandibular incisor, maxillary incisor, GoGn, occlusal plane angle, Frankfort horizontal plane (FHP), and CoA. Superimposition of structures seemed to influence the results more than magnification, and neither one of them significantly influenced the measurements. Considerable individual variability may occur, especially for mandibular and maxillary incisors, FHP and occlusal plane.

Aerobic endurance capacity affects spatial memory and SIRT1 is a potent modulator of 8-oxoguanine repair.

PubMed

Sarga, L; Hart, N; Koch, L G; Britton, S L; Hajas, G; Boldogh, I; Ba, X; Radak, Z

2013-11-12

Regular exercise promotes brain function via a wide range of adaptive responses, including the increased expression of antioxidant and oxidative DNA damage-repairing systems. Accumulation of oxidized DNA base lesions and strand breaks is etiologically linked to for example aging processes and age-associated diseases. Here we tested whether exercise training has an impact on brain function, extent of neurogenesis, and expression of 8-oxoguanine DNA glycosylase-1 (Ogg1) and SIRT1 (silent mating-type information regulation 2 homolog). To do so, we utilized strains of rats with low- and high-running capacity (LCR and HCR) and examined learning and memory, DNA synthesis, expression, and post-translational modification of Ogg1 hippocampal cells. Our results showed that rats with higher aerobic/running capacity had better spatial memory, and expressed less Ogg1, when compared to LCR rats. Furthermore, exercise increased SIRT1 expression and decreased acetylated Ogg1 (AcOgg1) levels, a post-translational modification important for efficient repair of 8-oxo-7,8-dihydroguanine (8-oxoG). Our data on cell cultures revealed that nicotinamide, a SIRT1-specific inhibitor, caused the greatest increase in the acetylation of Ogg1, a finding further supported by our other observations that silencing SIRT1 also markedly increased the levels of AcOgg1. These findings imply that high-running capacity is associated with increased hippocampal function, and SIRT1 level/activity and inversely correlates with AcOgg1 levels and thereby the repair of genomic 8-oxoG. Copyright © 2013 IBRO. Published by Elsevier Ltd. All rights reserved.

Abundance Trends and Status of the Little Colorado River Population of Humpback Chub: An Update Considering 1989-2006 Data

USGS Publications Warehouse

Coggins,, Lewis G.

2008-01-01

EXECUTIVE SUMMARY In 1967, the humpback chub (Gila cypha) (HBC) was added to the federal list of endangered species and is today protected under the Endangered Species Act of 1973. Only six populations of humpback chub are currently known to exist, five in the Colorado River Basin above Lees Ferry, Arizona, and one in Grand Canyon, Arizona. The majority of Grand Canyon humpback chub are found in the Little Colorado River (LCR)-the largest tributary to the Colorado River in Grand Canyon-and the Colorado River near its confluence with the Little Colorado River. Monitoring and research of the Grand Canyon humpback chub population is overseen by the U.S. Geological Survey's (USGS) Grand Canyon Monitoring and Research Center (GCMRC) under the auspices of the Glen Canyon Dam Adaptive Management Program (GCDAMP), a Federal initiative to protect and improve resources downstream of Glen Canyon Dam. This report provides updated information on the status and trends of the LCR population in light of new information and refined assessment methodology. An earlier assessment of the LCR population (Coggins and others, 2006a) used data collected during 1989?2002; the assessment provided here includes that data and additional data collected through 2006. Catch-rate indices, closed population mark-recapture model abundance estimates, results from the original age-structured mark recapture (ASMR) model (Coggins and others, 2006b), and a newly refined ASMR model are presented. This report also seeks to (1) formally evaluate alternative stock assessment models using Pearson residual analyses and information theoretic procedures, (2) use mark-recapture data to estimate the relationship between HBC age and length, (3) translate uncertainty in the assignment of individual fish age to resulting estimates of recruitment and abundance from the ASMR model, and (4) evaluate past and present stock assessments considering the available data sources and analyses, recognizing the limitations inherent in both. A major task of this study was to improve the overall methodology used to conduct HBC stock assessment by addressing concerns identified in an independent review conducted in 2003 (Kitchell and others, 2003). The review report identified that the current technique of assigning age to individual fish based on length was a potential source of bias in ASMR estimates of abundance and recruitment, and called for a more complete examination of this potential error source. Additionally, the review suggested that further work to develop procedures to better arbitrate among alternative assessment models (e.g., ASMR 1?3) would be beneficial. To address the first of the concerns identified by the independent review, this study uses mark-recapture data to develop a temperature-dependent growth model to characterize the relationship between HBC age and length. This model attempts to account for temperature differences resulting from both ontogenetic habitat shifts between the Little Colorado and the mainstem Colorado Rivers as well as seasonal variation in water temperature within the LCR. The resulting growth model is then used to characterize the error in assigning age to individual fish based on length. Results presented in this study suggest that ageing error does not result in large bias in either abundance or recruitment estimates from the ASMR model. However, incorporating ageing error into the assessment does result in less precise estimates, particularly for recruitment. To address the second concern brought forward in the review report related to model selection procedures, this study arbitrated among the competing models by both examining model fit using Pearson residual analyses and considering information theoretic measures. Although adult abundance estimates and trend varied little among all models considered, these procedures identified ASMR 3 as the model whose underlying assumptions were most consistent with the data. Because ASMR 3 is

CT Urography: Segmentation of Urinary Bladder using CLASS with Local Contour Refinement

PubMed Central

Cha, Kenny; Hadjiiski, Lubomir; Chan, Heang-Ping; Caoili, Elaine M.; Cohan, Richard H.; Zhou, Chuan

2016-01-01

Purpose We are developing a computerized system for bladder segmentation on CT urography (CTU), as a critical component for computer-aided detection of bladder cancer. Methods The presence of regions filled with intravenous contrast and without contrast presents a challenge for bladder segmentation. Previously, we proposed a Conjoint Level set Analysis and Segmentation System (CLASS). In case the bladder is partially filled with contrast, CLASS segments the non-contrast (NC) region and the contrast-filled (C) region separately and automatically conjoins the NC and C region contours; however, inaccuracies in the NC and C region contours may cause the conjoint contour to exclude portions of the bladder. To alleviate this problem, we implemented a local contour refinement (LCR) method that exploits model-guided refinement (MGR) and energy-driven wavefront propagation (EDWP). MGR propagates the C region contours if the level set propagation in the C region stops prematurely due to substantial non-uniformity of the contrast. EDWP with regularized energies further propagates the conjoint contours to the correct bladder boundary. EDWP uses changes in energies, smoothness criteria of the contour, and previous slice contour to determine when to stop the propagation, following decision rules derived from training. A data set of 173 cases was collected for this study: 81 cases in the training set (42 lesions, 21 wall thickenings, 18 normal bladders) and 92 cases in the test set (43 lesions, 36 wall thickenings, 13 normal bladders). For all cases, 3D hand segmented contours were obtained as reference standard and used for the evaluation of the computerized segmentation accuracy. Results For CLASS with LCR, the average volume intersection ratio, average volume error, absolute average volume error, average minimum distance and Jaccard index were 84.2±11.4%, 8.2±17.4%, 13.0±14.1%, 3.5±1.9 mm, 78.8±11.6%, respectively, for the training set and 78.0±14.7%, 16.4±16.9%, 18.2±15.0%, 3.8±2.3 mm, 73.8±13.4% respectively, for the test set. With CLASS only, the corresponding values were 75.1±13.2%, 18.7±19.5%, 22.5±14.9%, 4.3±2.2 mm, 71.0±12.6%, respectively, for the training set and 67.3±14.3%, 29.3±15.9%, 29.4±15.6%, 4.9±2.6 mm, 65.0±13.3%, respectively, for the test set. The differences between the two methods for all five measures were statistically significant (p<0.001) for both the training and test sets. Conclusions The results demonstrate the potential of CLASS with LCR for segmentation of the bladder. PMID:24801066

Effects of a test flood on fishes of the Colorado River in Grand Canyon, Arizona

USGS Publications Warehouse

Valdez, R.A.; Hoffnagle, T.L.; McIvor, C.C.; McKinney, T.; Leibfried, W.C.

2001-01-01

A beach/habitat-building flow (i.e., test flood) of 1274 m3/s, released from Glen Canyon Dam down the Colorado River through Grand Canyon, had little effect on distribution, abundance, or movement of native fishes, and only short-term effects on densities of some nonnative species Shoreline and backwater catch rates of native fishes, including juvenile humpback chub (Gila cypha), flannelmouth suckers (Catostomus latipinnis), and bluehead suckers (C. discobolus), and all ages of speckled dace (Rhinichthys osculus), were not significantly different before and after the flood. Annual spring spawning migrations of flannelmouth suckers into the Paria River and endangered humpback chub into the Little Colorado River (LCR) took place during and after the flood, indicating no impediment to fish migrations. Pre-spawning adults staged in large slack water pools formed at the mouths of these tributaries during the flood. Net movement and habitat used by nine radio-tagged adult humpback chub during the flood were not significantly different from prior observations. Diet composition of adult humpback chub varied, but total biomass did not differ significantly before, during, and after the flood, indicating opportunistic feeding for a larger array of available food items displaced by the flood. Numbers of nonnative rainbow trout (Oncorhynchus mykiss) <152 mm total length decreased by ???8% in electrofishing samples from the dam tailwaters (0-25 km downstream of the dam) during the flood. Increased catch rates in the vicinity of the LCR (125 km downstream of the dam) and Hell's Hollow (314 km downstream of the dam) suggest that these young trout were displaced downstream by the flood, although displacement distance was unknown since some fish could have originated from local populations associated with intervening tributaries. Abundance, catch rate, body condition, and diet of adult rainbow trout in the dam tailwaters were not significantly affected by the flood, and the flood did not detrimentally affect spawning success; catch of young-of-year increased by 20% in summer following the flood. Post-flood catch rates of nonnative fathead minnows (Pimephales promelas) in shorelines and backwaters, and plains killifish (Fundulus zebrinus) in backwaters decreased in the vicinity of the LCR, and fathead minnows increased near Hell's Hollow, suggesting that the flood displaced this nonnative species. Densities of rainbow trout and fathead minnows recovered to pre-flood levels eight months after the flood by reinvasion from tributaries and reproduction in backwaters. We concluded that the flood was of insufficient magnitude to substantially reduce populations of nonnative fishes, but that similar managed floods can disadvantage alien predators and competitors and enhance survival of native fishes.

Re-Introduction of Lower Columbia River Chum Salmon into Duncan Creek, 2002-2003 Annual Report.

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

Hillson, Todd D.

2003-10-15

The National Marine Fisheries Service (NMFS) listed Lower Columbia River chum as threatened under the auspices of the Endangered Species Act (ESA) in March of 1999 (64 FR 14508, March 25, 1999). The listing was in response to reduction in abundance from historical levels of more than half a million returning adults to fewer than 10,000 present day spawners. Harvest, loss of habitat, changes in flow regimes, riverbed movement and heavy siltation have been largely responsible for the decline of Columbia River chum salmon. The timing of seasonal changes in river flow and water temperatures is perhaps the most criticalmore » factor in structuring the freshwater life history of this species. This is especially true of the population located directly below Bonneville Dam where hydropower operations can block access to spawning sites, dewater redds, strand fry, cause scour or fill of redds and increase sedimentation of spawning gravels. Currently, only two main populations are recognized as genetically distinct in the Columbia River, although spawning has been documented in most lower Columbia River tributaries. The first is located in the Grays River (RKm 34) (Grays population), a tributary of the Columbia, and the second is a group of spawners that utilize the Columbia River just below Bonneville Dam (RKm 235) adjacent to Ives Island and in Hardy and Hamilton creeks (Lower Gorge population). A possible third population of mainstem spawners, found in the fall of 1999, were located spawning above the I-205 bridge (approximately RKm 182), this aggregation is referred to as the Woods Landing/Rivershore population or the I-205 group. The recovery strategy for Lower Columbia River (LCR) chum as outlined in Hatchery Genetic Management Plans (HGMP) has three main tasks. First, determine if remnant populations of LCR chum salmon exist in LCR tributaries. Second, if such populations exist, develop stock-specific recovery plans involving habitat restoration including the creation of spawning refugias, supplementation if necessary and a habitat and fish monitoring and evaluation plan. If chum have been extirpated from previously utilized streams, develop re-introduction plans that utilize appropriate genetic donor stock(s) of LCR chum salmon and integrate habitat improvement and fry-to-adult survival evaluations. Third, reduce extinction risks to the Grays River chum salmon population by randomly capturing adults in the basin for use in a supplementation program and reintroduction into the Chinook River basin. The Duncan Creek project was developed using the same recovery strategy implemented for LCR chum. Biologists with the WDFW and Pacific States Marine Fisheries Commission (PSMFC) identified Duncan Creek as an ideal upriver location below Bonneville Dam for chum re-introduction. It has several attributes that make it a viable location for a re-introduction project: historically chum salmon were present, the creek is low gradient, has numerous springs/seeps, has a low potential for future development and is located close to a donor population of Lower Gorge chum. The Duncan Creek project has two goals: (1) re-introduction of chum into Duncan Creek by providing off channel high-quality spawning and incubation areas, and (2) to simultaneously evaluate natural recolonization and a supplementation strategy where adults are collected and spawned artificially at a hatchery. For supplementation, eggs are incubated and the fry reared at the Washougal Hatchery to be released back into Duncan Creek. The tasks associated with re-establishing a naturally self-sustaining population include: (1) removing mud, sand and organics present in four of the creek branches and replace with gravels expected to provide maximum egg-to-fry survival rates to a depth of at least two feet; (2) armoring the sides of these channels to reduce importation of sediment by fish spawning on the margins; (3) planting native vegetation adjacent to the channels to stabilize the banks, trap silt and provide shade; (4) annual sampling of gravel in the spawning channels to detect changes in gravel composition and sedimentation levels. Tasks associated with the second goal of the recovery strategy for Lower Columbia River chum are detailed in The Monitoring and Evaluation Plan for the Duncan Creek Chum Salmon Reintroduction Program. Four main questions are used to evaluate the success of this program: (1) what egg-to-fry survival rates are being achieved in the renovated channels, (2) what is the survival of the eggs and fry used in the artificial rearing program in Duncan Creek, (3) what is the survival and spawning ground distribution of adult chum salmon produced from the spawning channels and the artificial rearing program, and (4) what is the straying rate of non-program chum salmon into Duncan Creek. The monitoring portion of the Duncan M&E includes documenting and monitoring the physical attributes of the channels.« less

Contribution of regional brain melanocortin receptor subtypes to elevated activity energy expenditure in lean, active rats

PubMed Central

Shukla, Charu; Koch, Lauren G.; Britton, Steven L.; Cai, Minying; Hruby, Victor J.; Bednarek, Maria; Novak, Colleen M.

2015-01-01

Physical activity and non-exercise activity thermogenesis (NEAT) are crucial factors accounting for individual differences in body weight, interacting with genetic predisposition. In the brain, a number of neuroendocrine intermediates regulate food intake and energy expenditure (EE); this includes the brain melanocortin (MC) system, consisting of melanocortin peptides as well as their receptors (MCR). MC3R and MC4R have emerged as critical modulators of EE and food intake. To determine how variance in MC signaling may underlie individual differences in physical activity levels, we examined behavioral response to MC receptor agonists and antagonists in rats that show high and low levels of physical activity and NEAT, that is, high- and low-capacity runners (HCR, LCR), developed by artificial selection for differential intrinsic aerobic running capacity. Focusing on the hypothalamus, we identified brain region-specific elevations in expression of MCR 3, 4, and also MC5R, in the highly active, lean HCR relative to the less active and obesity-prone LCR. Further, the differences in activity and associated EE as a result of MCR activation or suppression using specific agonists and antagonists were similarly region-specific and directly corresponded to the differential MCR expression patterns. The agonists and antagonists investigated here did not significantly impact food intake at the doses used, suggesting that the differential pattern of receptor expression may by more meaningful to physical activity than to other aspects of energy balance regulation. Thus, MCR-mediated physical activity may be a key neural mechanism in distinguishing the lean phenotype and a target for enhancing physical activity and NEAT. PMID:26404873

The golden proportion in facial soft-tissues of Vietnamese females.

PubMed

Nguyen, Minh Son; Saag, Mare; Le, Van Nho; Nguyen, Thuy Trang; Nguyen, Bui Bao Tien; Jagomägi, Triin

2016-01-01

Objective of the current study was to evaluate proportions of frontal facial soft-tissues of Vietnamese females correspond to the golden proportion (GP). Sixty frontal facial photographs of Vietnamese female students aged 19 years were selected. The selected participants had symmetrical face, a Class I relationship occlusion, complete lip closure, and no history of trauma and orthodontic treatment. The photographic record was set-up with a white backdrop, fill light, a reflector, a camera Canon 650D and subjects were asked to sit in a standard position. Trichion (TR), Temporal soft tissue (TS), Lateral canthus (LC), Lateral nasal (LN), Chilion (CH), and Menton (ME) point were used for photometric measurements on CorelDRAW Graphic X3 software. The vertical facial proportions (mean, percentage compared with GP) were significantly higher than the GP including: LC-CH:CH-ME (1.661, 102%), LN-ME:LC-LN (1.729, 106%), TR-ME:LC-ME (1.739, 107%), TR-LN:LN-ME (1.759, 108%). Whereas TR-ME:LN-ME; CH-ME:LN-CH; LC-LN:LN-CH had ratios of 84% - 92% and were significantly smaller in comparison with GP. The horizontal facial proportions of CHR-L:LNR-L, LCR-L:CHR-L and TSR-L:LCR-L deviated from the GP with mean values of 1.221, 1.922 and 1.229, respectively. The new mean values of TR-ME:LC-ME (1.733), TR-LN:LN-ME (1.732), LC-ME:TR-LC (1.374), CH-ME:LN-CH (1.524), which were predictable changes in proportions, can be significantly converged to the GP if LC-CH:CH-ME is equal to the GP. Soft-tissue facial proportions of Vietnamese females did not correspond to the GP. Changing the lower third face may create harmony vertical facial proportions.

Comparison of the Effectiveness of Radiofrequency Ablation With Stereotactic Body Radiation Therapy in Inoperable Stage I Non-Small Cell Lung Cancer: A Systemic Review and Pooled Analysis.

PubMed

Bi, Nan; Shedden, Kerby; Zheng, Xiangpeng; Kong, Feng-Ming Spring

2016-08-01

To performed a systematic review and pooled analysis to compare clinical outcomes of stereotactic body radiation therapy (SBRT) and radiofrequency ablation (RFA) for the treatment of medically inoperable stage I non-small cell lung cancer. A comprehensive literature search for published trials from 2001 to 2012 was undertaken. Pooled analyses were performed to obtain overall survival (OS) and local tumor control rates (LCRs) and adverse events. Regression analysis was conducted considering each study's proportions of stage IA and age. Thirty-one studies on SBRT (2767 patients) and 13 studies on RFA (328 patients) were eligible. The LCR (95% confidence interval) at 1, 2, 3, and 5 years for RFA was 77% (70%-85%), 48% (37%-58%), 55% (47%-62%), and 42% (30%-54%) respectively, which was significantly lower than that for SBRT: 97% (96%-98%), 92% (91%-94%), 88% (86%-90%), and 86% (85%-88%) (P<.001). These differences remained significant after correcting for stage IA and age (P<.001 at 1 year, 2 years, and 3 years; P=.04 at 5 years). The effect of RFA was not different from that of SBRT on OS (P>.05). The most frequent complication of RFA was pneumothorax, occurring in 31% of patients, whereas that for SBRT (grade ≥3) was radiation pneumonitis, occurring in 2% of patients. Compared with RFA, SBRT seems to have a higher LCR but similar OS. More studies with larger sample sizes are warranted to validate such findings. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

NASBA: A detection and amplification system uniquely suited for RNA

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

Sooknanan, R.; Malek, L.T.

1995-06-01

The invention of PCR (polymerase chain reaction) has revolutionized our ability to amplify and manipulate a nucleic acid sequence in vitro. The commercial rewards of this revolution have driven the development of other nuclei acid amplification and detection methodologies. This has created an alphabet soup of technologies that use different amplification methods, including NASBA (nucleic acid sequence-based amplification), LCR (ligase chain reaction), SDA (strand displacement amplification), QBR (Q-beta replicase), CPR (cycling probe reaction), and bDNA (branched DNA). Despite the differences in their processes, these amplification systems can be separated into two broad categories based on how they achieve their goal:more » sequence-based amplification systems, such as PCR, NASBA, and SDA, amplify a target nucleic acid sequence. Signal-based amplification systems, such as LCR, QBR, CPR and bDNA, amplify or alter a signal from a detection reaction that is target-dependent. While the various methods have relative strengths and weaknesses, only NASBA offers the unique ability to homogeneously amplify an RNA analyte in the presence of homologous genomic DNA under isothermal conditions. Since the detection of RNA sequences almost invariably measures biological activity, it is an excellent prognostic indicator of activities as diverse as virus production, gene expression, and cell viability. The isothermal nature of the reaction makes NASBA especially suitable for large-scale manual screening. These features extend NASBA`s application range from research to commercial diagnostic applications. Field test kits are presently under development for human diagnostics as well as the burgeoning fields of food and environmental diagnostic testing. These developments suggest future integration of NASBA into robotic workstations for high-throughput screening as well. 17 refs., 1 tab.« less

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

Dobrakowski, Michał, E-mail: michal.dobrakowski@po

The aim of the study was to investigate the influence of a short-term exposure to lead on the blood morphology and the levels of selected cytokines related to hematopoiesis in occupationally exposed workers. The study population included 37 males occupationally exposed to lead for 36 to 44 days. Their blood lead level raised from 10.7 ± 7.67 μg/dl at baseline to the level of 49.1 ± 14.1 μg/dl at the end of the study. The level of hemoglobin and values of MCH and MCHC were decreased due to a short-term exposure to lead by 2%, 2%, and 1%, respectively. Themore » counts of WBC, LYM, and MXD increased significantly by 5%, 7%, and 35%. Similarly, the count of PLT increased by 7%, while PDW, MPV, and P-LCR decreased by 6%, 3%, and 9%, respectively. The levels of IL-7, G-CSF, HGF, PDGF AB/BB, SCF, and PECAM-1, decreased significantly by 30%, 33%, 8%, 30%, 25%, and 20%, respectively. A short-term occupational exposure to lead results in a decreased hemoglobin level and increased counts of WBC and PLT. Changes in counts and proportions of different types of leukocytes and decreased values of PLT indices, such as PDW, MPV, and P-LCR, due to the subacute lead-exposure may be associated with lead-induced decreased levels of cytokines related to hematopoiesis, including SCF, G-CSF, IL-7, and PDGF. - Highlights: • Subacute exposure to lead results in a decreased hemoglobin level. • Subacute exposure to lead results in increased counts of WBC and PLT. • Subacute exposure to lead decreases the levels of SCF, G-CSF, IL-7, and PDGF.« less

T cells play an essential role in anti-F1 mediated rapid protection against bubonic plague.

PubMed

Levy, Yinon; Flashner, Yehuda; Tidhar, Avital; Zauberman, Ayelet; Aftalion, Moshe; Lazar, Shirley; Gur, David; Shafferman, Avigdor; Mamroud, Emanuelle

2011-09-16

Plague, which is initiated by Yersinia pestis infection, is a fatal disease that progresses rapidly and leads to high mortality rates if not treated. Antibiotics are an effective plague therapy, but antibiotic-resistant Y. pestis strains have been reported and therefore alternative countermeasures are needed. In the present study, we assessed the potential of an F1 plus LcrV-based vaccine to provide protection shortly pre- or post-exposure to a lethal Y. pestis infection. Mice vaccinated up to one day before or even several hours after subcutaneous challenge were effectively protected. Mice immunized one or three days pre-challenge were protected even though their anti-F1 and anti-LcrV titers were below detection levels at the day of challenge. Moreover, using B-cell deficient μMT mice, we found that rapidly induced protective immunity requires the integrity of the humoral immune system. Analysis of the individual contributions of vaccine components to protection revealed that rF1 is responsible for the observed rapid antibody-mediated immunity. Applying anti-F1 passive therapy in the mouse model of bubonic plague demonstrated that anti-F1 F(ab')(2) can delay mortality, but it cannot provide long-lasting protection, as do intact anti-F1 molecules. Fc-dependent immune components, such as the complement system and (to a lesser extent) neutrophils, were found to contribute to mouse survival. Interestingly, T cells but not B cells were found to be essential for the recovery of infected animals following passive anti-F1 mediated therapy. These data extend our understanding of the immune mechanisms required for the development of a rapid and effective post-exposure therapy against plague. Copyright © 2011 Elsevier Ltd. All rights reserved.

On-line preconcentration and speciation of arsenic by flow injection hydride generation atomic absorption spectrophotometry.

PubMed

Narcise, Cristine Ingrid S; Coo, Lilibeth Dlc; Del Mundo, Florian R

2005-12-15

A flow injection-column preconcentration-hydride generation atomic absorption spectrophotometric (FI-column-HGAAS) method was developed for determining mug/l levels of As(III) and As(V) in water samples, with simultaneous preconcentration and speciation. The speciation scheme involved determining As(V) at neutral pH and As(III+V) at pH 12, with As(III) obtained by difference. The enrichment factor (EF) increased with increase in sample loading volume from 2.5 to 10ml, and for preconcentration using the chloride-form anion exchange column, EFs ranged from 5 to 48 for As(V) and 4 to 24 for As(III+V), with corresponding detection limits of 0.03-0.3 and 0.07-0.3mug/l. Linear concentration range (LCR) also varied with sample loading volume, and for a 5-ml sample was 0.3-5 and 0.2-8mug/l for As(V) and As(III+V), respectively. Sample throughput, which decreased with increase in sample volume, was 8-17 samples/h. For the hydroxide-form column, the EFS for 2.5-10ml samples were 3-23 for As(V) and 2-15 for As(III+V), with corresponding detection limits of 0.07-0.4 and 0.1-0.5mug/l. The LCR for a 5-ml sample was 0.3-10mug/l for As(V) and 0.2-20mug/l for As(III+V). Sample throughput was 10-20 samples/h. The developed method has been effectively applied to tap water and mineral water samples, with recoveries ranging from 90 to 102% for 5-ml samples passed through the two columns.

Comparison of the Effectiveness of Radiofrequency Ablation With Stereotactic Body Radiation Therapy in Inoperable Stage I Non-Small Cell Lung Cancer: A Systemic Review and Pooled Analysis

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

Bi, Nan; Department of Radiation Oncology, Cancer Hospital and Institute, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing; Shedden, Kerby

Purpose: To performed a systematic review and pooled analysis to compare clinical outcomes of stereotactic body radiation therapy (SBRT) and radiofrequency ablation (RFA) for the treatment of medically inoperable stage I non-small cell lung cancer. Methods and Materials: A comprehensive literature search for published trials from 2001 to 2012 was undertaken. Pooled analyses were performed to obtain overall survival (OS) and local tumor control rates (LCRs) and adverse events. Regression analysis was conducted considering each study's proportions of stage IA and age. Results: Thirty-one studies on SBRT (2767 patients) and 13 studies on RFA (328 patients) were eligible. The LCR (95%more » confidence interval) at 1, 2, 3, and 5 years for RFA was 77% (70%-85%), 48% (37%-58%), 55% (47%-62%), and 42% (30%-54%) respectively, which was significantly lower than that for SBRT: 97% (96%-98%), 92% (91%-94%), 88% (86%-90%), and 86% (85%-88%) (P<.001). These differences remained significant after correcting for stage IA and age (P<.001 at 1 year, 2 years, and 3 years; P=.04 at 5 years). The effect of RFA was not different from that of SBRT on OS (P>.05). The most frequent complication of RFA was pneumothorax, occurring in 31% of patients, whereas that for SBRT (grade ≥3) was radiation pneumonitis, occurring in 2% of patients. Conclusions: Compared with RFA, SBRT seems to have a higher LCR but similar OS. More studies with larger sample sizes are warranted to validate such findings.« less

Occurrence of commonly used pesticides in personal air samples and their associated health risk among paddy farmers.

PubMed

Hamsan, Hazwanee; Ho, Yu Bin; Zaidon, Siti Zulfa; Hashim, Zailina; Saari, Nazamid; Karami, Ali

2017-12-15

Tanjung Karang, Selangor, is widely known for its paddy cultivation activity and hosts the third largest paddy field in Malaysia. Pesticides contamination in agriculture fields has become an unavoidable problem, as pesticides are used to increase paddy productivity and reduce plant disease. Human exposure to agrichemicals is common and could results in both acute and chronic health effects, such as acute and chronic neurotoxicity. This study aims to determine the concentrations of commonly used pesticides (azoxystrobin, buprofezin, chlorantraniliprole, difenoconazole, fipronil, imidacloprid, isoprothiolane, pretilachlor, propiconazole, pymetrozine, tebuconazole, tricyclazole, and trifloxystrobin) in personal air samples and their associated health risks among paddy farmers. Eighty-three farmers from Tangjung Karang, Selangor were involved in this study. A solid sorbent tube was attached to the farmer's breathing zone with a clip, and an air pump was fastened to the belt to collect personal air samples. Pesticides collected in the XAD-2 resin were extracted with acetone, centrifuged, concentrated via nitrogen blowdown and reconstituted with 1mL of 3:1 ultrapure water/HPLC-grade methanol solution. The extract was analyzed using ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). The target compounds were detected with a maximum concentration reaching up to 462.5ngm -3 (fipronil). The hazard quotient (HQ) was less than 1 and the hazard index (HI) value was 3.86×10 -3 , indicating that the risk of pesticides related diseases was not significant. The lifetime cancer risk (LCR) for pymetrozine was at an acceptable level (LCR<10 -6 ) with 4.10×10 -8 . The results reported in this study can be beneficial in terms of risk management within the agricultural community. Copyright © 2017 Elsevier B.V. All rights reserved.

Investigation of a new in-line leukocyte reduction filter for packed red blood cells.

PubMed

Mönninghoff, J; Moog, R

2012-06-01

Occasionally there are adverse transfusion reactions in the therapeutic use of packed red blood cells. Some of those reactions are caused by the presence of white blood cells (WBCs). Both immunogenic and infectious transfusion reactions are significantly influenced by the level of white blood cell contamination. The flexible in-line red cell filtration system Leucoflex LCR Diamond (Macopharma) was investigated. According to manufacturer information the system has a smaller filter surface (46 cm(2)) than the previous filter LCR-5 (53 cm(2)). Main difference with the previous model is the rhomboid design. The filter tube connections are not at the level of the centre edge, but at two opposite corners. Eighteen red cell concentrates were produced under Good Manufacturing Practice conditions in routine operation. To ensure the quality of the filter system every 7 days metabolic parametres such as WBC count, haemoglobin content, haemolysis rate, potassium load, pH and ATP content were analysed over a storage period of 49 days. The mean product volume was 260.7 mL after filtration. Average haemoglobin content was 51.8 g per unit and WBC contamination was 0.02 × 10(6)per unit. Haemolysis rate was 0.05% directly after filtration and 0.20% at the end of storage. Immediately after filtration the potassium concentration was 1.3 mmol/L and the pH was 7.37. During whole storage time the ATP level was maintained above 2.0 μmol per g haemoglobin. The tested filtration system is suitable for quality-assured production of red blood cell concentrates meeting national and international guidelines. Copyright © 2012 Elsevier Ltd. All rights reserved.

Epidemiologic analysis of a sarcoid outbreak involving 12 of 111 donkeys in Northern Italy.

PubMed

Abel-Reichwald, Hans; Hainisch, Edmund K; Zahalka, Sophie; Corteggio, Annunziata; Borzacchiello, Giuseppe; Massa, Barbara; Merlone, Luca; Nasir, Lubna; Burden, Faith; Brandt, Sabine

2016-11-30

Equine sarcoids develop upon bovine papillomavirus type 1 or 2 (BPV1, BPV2) infection in conjunction with trauma and represent the most common tumour disease in horses and other equids, including donkeys. In face of a sarcoid outbreak involving 12 of 111 donkeys and mules at the 'Rifugio degli Asinelli', a subsidiary charity organization of The Donkey Sanctuary, non-invasively collected sample material including crusts, dandruff, swabs and hair roots was collected from sarcoid-affected and 26 healthy donkeys, as well as dandruff from a grooming kit and tabanids caught from or in the vicinity of sarcoid patients. In addition five previously collected sarcoids stored in formalin were provided. DNA isolated from collected material was tested for the presence of the BPV1/2 E5 oncogene using PCR. Positive samples were further analysed by E2/E4 and LCR PCR and amplicon sequencing to determine a possible common source of infection via comparative alignment of intralesional BPV1/2 gene variants. IC/PCR was used to assess sample aliquots for the presence of BPV1/2 virions, and IHC to analyse five tumours for BPV1 E5 and L1 protein expression. All sarcoid-affected donkeys, two of 55 tabanids and dandruff from a curry comb tested positive for BPV1/2 E5, yet negative by IC/PCR. Healthy animals were BPV1/2-free. IHC revealed different levels of intralesional E5 and L1 expression. A series of BPV1 E5, E2, and LCR variants and BPV2 E5 were detected from donkeys, indicating that they had accidently developed sarcoids at about the same time rather than having acquired disease from each other. Copyright © 2016 Elsevier B.V. All rights reserved.

Characterization and health risk assessment of VOCs in occupational environments in Buenos Aires, Argentina

NASA Astrophysics Data System (ADS)

Colman Lerner, J. E.; Sanchez, E. Y.; Sambeth, J. E.; Porta, A. A.

2012-08-01

To detect volatile organic compounds (VOCs) in indoor air in small enterprises in La Plata city and surrounding areas, sampling was conducted using passive diffusion monitors (3M-3500) and analysis of the samples were performed byCG-FID. Analytic methodology was optimized for 23 VOCs (n-alkanes, cycloalkanes, aromatic and chlorinated compounds, ketones and terpenes compounds) by determining the recovery factor and detection limit for each analyte. Different recovery values were obtained by desorbing with a mixture of dichloromethane: methanol (50:50), with a standard deviation lower than 5%. Enterprise analyzed included chemical analysis laboratories, sewing workrooms, electromechanical repair and car painting centers, take away food shops, and a photocopy center. The highest levels of VOCs were found to be in electromechanical repair and car painting centers (hexane, BTEX, CHCl3, CCl4) followed by chemical analysis laboratories and sewing workrooms. Cancer and noncancer risks were assessed using conventional approaches (HQ and LCR, US EPA) using the benzene, trichloroethylene, chloroform for cancer risk, and toluene, xylene and n-hexane, for noncancer risks as markers. The results showed different LCR for benzene and trichloroethylene between the different indoor environments analyzed (electromechanical repair and car painting center ≫ others) and chloroform (laboratory > others), but comparing with the results obtained by other research, are in similar order of magnitude for equivalents activities. Similar finding were founded for HQ. Comparing these results with the worker protection legislation the electromechanical repair and car painting center and chemical analysis laboratories are close to the limits advised by OSHA and ACGIH. These facts show the importance of the use of abatement technologies for the complete reduction of VOCs levels, to mitigate their impact in the worker's health and their venting to the atmosphere.

Selection-, age-, and exercise-dependence of skeletal muscle gene expression patterns in a rat model of metabolic fitness.

PubMed

Ren, Yu-Yu; Koch, Lauren G; Britton, Steven L; Qi, Nathan R; Treutelaar, Mary K; Burant, Charles F; Li, Jun Z

2016-11-01

Intrinsic aerobic exercise capacity can influence many complex traits including obesity and aging. To study this connection we established two rat lines by divergent selection of untrained aerobic capacity. After 32 generations the high capacity runners (HCR) and low capacity runners (LCR) differed in endurance running distance and body fat, blood glucose, other health indicators, and natural life span. To understand the interplay among genetic differences, chronological age, and acute exercise we performed microarray-based gene expression analyses in skeletal muscle with a 2×2×2 design to simultaneously compare HCR and LCR, old and young animals, and rest and exhaustion. Transcripts for mitochondrial function are expressed higher in HCRs than LCRs at both rest and exhaustion and for both age groups. Expression of cell adhesion and extracellular matrix genes tend to decrease with age. This and other age effects are more prominent in LCRs than HCRs, suggesting that HCRs have a slower aging process and this may be partly due to their better metabolic health. Strenuous exercise mainly affects transcription regulation and cellular response. The effects of any one factor often depend on the other two. For example, there are ∼140 and ∼110 line-exercise "interacting" genes for old and young animals, respectively. Many genes highlighted in our study are consistent with prior reports, but many others are novel. The gene- and pathway-level statistics for the main effects, either overall or stratified, and for all possible interactions, represent a rich reference dataset for understanding the interdependence among lines, aging, and exercise. Copyright © 2016 the American Physiological Society.

Regulation of the Yersinia type III secretion system: traffic control

PubMed Central

Dewoody, Rebecca S.; Merritt, Peter M.; Marketon, Melanie M.

2013-01-01

Yersinia species, as well as many other Gram-negative pathogens, use a type III secretion system (T3SS) to translocate effector proteins from the bacterial cytoplasm to the host cytosol. This T3SS resembles a molecular syringe, with a needle-like shaft connected to a basal body structure, which spans the inner and outer bacterial membranes. The basal body of the injectisome shares a high degree of homology with the bacterial flagellum. Extending from the T3SS basal body is the needle, which is a polymer of a single protein, YscF. The distal end of the needle serves as a platform for the assembly of a tip complex composed of LcrV. Though never directly observed, prevailing models assume that LcrV assists in the insertion of the pore-forming proteins YopB and YopD into the host cell membrane. This completes a bridge between the bacterium and host cell to provide a continuous channel through which effectors are delivered. Significant effort has gone into understanding how the T3SS is assembled, how its substrates are recognized and how substrate delivery is controlled. Arguably the latter topic is the least understood; however, recent advances have provided new insight, and therefore, this review will focus primarily on summarizing the current state of knowledge regarding the control of substrate delivery by the T3SS. Specifically, we will discuss the roles of YopK, as well as YopN and YopE, which have long been linked to regulation of translocation. We also propose models whereby the YopK regulator communicates with the basal body of the T3SS to control translocation. PMID:23390616

The evolution of analgesia in an 'accelerated' recovery programme for resectional laparoscopic colorectal surgery with anastomosis.

PubMed

Zafar, N; Davies, R; Greenslade, G L; Dixon, A R

2010-02-01

The study set out to analyse the outcomes of an evolving accelerated recovery programme after laparoscopic colorectal resection (LCR). The results of a prospective electronic database (March 2000 - April 2008) were analysed. There were 353 consecutive patients undergoing 'three port' high anterior resection (AR) (237 without covering stoma) and 166 a right hemicolectomy (RHC). One hundred thirty-eight had postoperative analgesia using paracetamol IV and oral analgesia (IVP); 27 (16.3%) received additional parenteral morphine and were excluded. Patient controlled morphine analgesia (PCA) was used in 138. Transversus abdominis plane (TAP) blocks, supplemented by IV paracetamol and oral analgesia were used in the last 50 patients. The time to the resumption of diet was significantly reduced with TAP analgesia (median 12 h) and IVP (median 12 h) compared with PCA median (36 h) (chi(2) = 143; 4df: P < 0.001). The postoperative hospital stay was significantly reduced with TAP analgesia (median 2 days) and IVP (median 3 days) compared with PCA (median 5 days); chi(2) = 73; 2df: P < 0.001. Seventeen (34%) TAP and nine (6.5%) IVP patients were discharged within 24 h of surgery compared with no patient in the PCA group. Ninety-three per cent of PCA, 35% IVP and 10% TAP patients were discharged in more than 3 days. The movement towards 'accelerated recovery' was not associated with any increased risk of urinary retention, return to theatre, readmission and/or 30 day mortality. Laparoscopic surgery utilizing IV paracetamol and TAP blocks for postoperative analgesia aids safe effective 'accelerated recovery' in an unselected patient population undergoing right hemicolectomy and high anterior resection. Routine epidural anaesthesia is unnecessary for LCR. Morphine PCA is associated with delayed recovery.

Synchronization of a Josephson junction array in terms of global variables

NASA Astrophysics Data System (ADS)

Vlasov, Vladimir; Pikovsky, Arkady

2013-08-01

We consider an array of Josephson junctions with a common LCR load. Application of the Watanabe-Strogatz approach [Physica DPDNPDT0167-278910.1016/0167-2789(94)90196-1 74, 197 (1994)] allows us to formulate the dynamics of the array via the global variables only. For identical junctions this is a finite set of equations, analysis of which reveals the regions of bistability of the synchronous and asynchronous states. For disordered arrays with distributed parameters of the junctions, the problem is formulated as an integro-differential equation for the global variables; here stability of the asynchronous states and the properties of the transition synchrony-asynchrony are established numerically.

Effect of aluminium substitution on the electrical properties of Ni-Zn nanoferrites

NASA Astrophysics Data System (ADS)

Paramesh, D.; Vijaya Kumar, K.; Venkat Reddy, P.

2017-12-01

Nanoferrites of general formula Ni0.5 Zn0.5 Alx Fe2-x O4 (x = 0.0, 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, 1.4, 1.6, 1.8 and 2.0) synthesized by sol-gel auto combustion technique can be characterized by dielectric behaviour and AC conductivity studies with the help of LCR impedance meter. This paper gives an insight on variations in dielectric constant, dielectric loss with reference to frequency, temperature and Al3+ ion substitution and also the determination of DC resistivity, activation energy and Curie temperature by two probe experimental set-up.

Frequency dispersion of capacitance-voltage characteristics in wide bandgap semiconductor-electrolyte junctions

NASA Astrophysics Data System (ADS)

Frolov, D. S.; Zubkov, V. I.

2016-12-01

The frequency dispersion of capacitance-voltage characteristics and derived charge carrier concentration with application to the junction between an electrolyte and wide band-gap semiconductors are investigated. To expand the measurement frequency range, the precision LCR-meter Agilent E4980A was connected to the electrochemical cell ECVPro Nanometrics via a specially designed switch unit. The influence of series resistance and degree of dopant ionization on the frequency dispersion of CV-measured characteristics are discussed. It was shown that in wide band-gap semiconductors one can get both total and ionized dopant concentration, depending on the test frequency choice for capacitance measurements.

Linearity and reproducibility response of Fricke dosimetry for low energy X-Ray beam

NASA Astrophysics Data System (ADS)

Mantuano, A.; de Amorim, G. J.; David, M. G.; Rosado, P. H. G.; Salata, C.; Magalhães, L. A. G.; deAlmeida, C. E.

2018-03-01

The Fricke dosimeter is the most used, liquid chemical dosimeter. It has been shown to be a feasible option for the absorbed dose standard. The present work aims to determinate a dose-response curve of Fricke solution using different doses and reproducibility test comparing the calculated dose to Fricke solution and Ionizing Chamber. Tests were performed using an X-ray irradiator for biological research at Radiological Science Laboratory (LCR/UERJ). The results showed a linear response to different doses of type A uncertainties from 0.08 to 1.2%. Reproducibility test showed type A uncertainties of 0.16% to the dosimeter.

Characterization of chitosan-starch blend based biopolymer electrolyte doped with ammonium nitrate

NASA Astrophysics Data System (ADS)

Shaffie, Ahmad Hakimi; Khiar, Azwani Sofia Ahmad

2018-06-01

Polymer electrolyte is an ionic conductor formed by dissolving salt in polymer host. In this work, starch/chitosan blend based polymer electrolyte was prepared with different weight percentage of Ammonium Nitrate (NH4NO3) via solution casting technique. The film was characterized by impedance spectroscopy HIOKI 3531- 01 LCR Hi-Tester to measure its ionic conductivity over a wide range of frequency between 50Hz-5MHz and at ambient temperature. Sample with 35 wt% of NH4NO3 shows the highest conductivity of (6.34 ± 1.52) = 10-7 Scm-1. X-ray diffraction (XRD) and Fourier transform infrared (FTIR) spectroscopy were used to correlate the ionic conductivity results.

Development of effective power supply using electric double layer capacitor for static magnetic field coils in fusion plasma experiments.

PubMed

Inomoto, M; Abe, K; Yamada, T; Kuwahata, A; Kamio, S; Cao, Q H; Sakumura, M; Suzuki, N; Watanabe, T; Ono, Y

2011-02-01

A cost-effective power supply for static magnetic field coils used in fusion plasma experiments has been developed by application of an electric double layer capacitor (EDLC). A prototype EDLC power supply system was constructed in the form of a series LCR circuit. Coil current of 100 A with flat-top longer than 1 s was successfully supplied to an equilibrium field coil of a fusion plasma experimental apparatus by a single EDLC module with capacitance of 30 F. The present EDLC power supply has revealed sufficient performance for plasma confinement experiments whose discharge duration times are an order of several seconds.

Dielectric and optical study of poly (methyl methacrylate) (PMMA) / Fe2O3 films

NASA Astrophysics Data System (ADS)

Anita, Chimankar, O. P.; Bansod, A. R.; Sannakki, Basavaraja

2013-06-01

Organic/inorganic polymer composite films containing poly (methyl-methacrylate) (PMMA)/ ferric oxide Fe2O3 were prepared following solution casting technique. Dielectric Properties of films has been studied using LCR meter at room temperature 26°C. Also optical properties have been studied using digital abbey refractometer. The dielectric behavior of films have been studied as a function of concentration, and at lower frequencies over the range 100 Hz-25 KHz, The results elucidate that 70:30 and 50:50 wt% of PMMA/Fe2O3 composite films posses optimal conducting properties due to observed electronic polarisability dip at 40Wt% of Fe2O3.

Inverse Trilateration - A Positional Goal Seeking Algorithm for a Swarm of Autonomous Drones

DTIC Science & Technology

2014-06-12

uny ouc or the n clrmus, \\Vt: ca11 •lcvc lop a system of equations for that ptnticular drmu .. ’- thc onn which will IJc att.<~mpting t.o fi11d u...a111l thc at’Tial vchich:. One sc:euerio for f’lctcnuiuing clistaocc to an nbject of unkuown location is to co1is11lcr a trauSJnith’r of lowvm powcr1...U1ing On6owU V"ual ond foertiol SvuinJ. In rroc. or the \\\\Orbhop Oh Visual Control of Mobilr Robols (VlC4’>.MoR) at thc - JEEE/RJS lntrmaiion"I Confl

Capacitance-digital and impedance converter as electrical tomography measurement system for biological tissue

NASA Astrophysics Data System (ADS)

Ikhsanti, Mila Izzatul; Bouzida, Rana; Wijaya, Sastra Kusuma; Rohmadi, Muttakin, Imamul; Taruno, Warsito P.

2017-02-01

This research aims to explore the feasibility of capacitance-digital converter and impedance converter for measurement module in electrical capacitance tomography (ECT) system. ECT sensor used was a cylindrical sensor having 8 electrodes. Absolute capacitance measurement system based on Sigma Delta Capacitance-to-Digital-Converter AD7746 has been shown to produce measurement with high resolution. Whereas, capacitance measurement with wide range of frequency is possible using Impedance Converter AD5933. Comparison of measurement accuracy by both AD7746 and AD5933 with reference of LCR meter was evaluated. Biological matters represented in water and oil were treated as object reconstructed into image using linear back projection (LBP) algorithm.

Dielectric spectroscopy of SiO2, ZnO - nanoparticle loaded epoxy resin in the frequency range of 20 Hz to 2 MHz

NASA Astrophysics Data System (ADS)

Thakor, Sanketsinh; Rana, V. A.; Vankar, H. P.

2017-05-01

In present work, Bisphenol A-(epichlorhydrin); epoxy resin with hardener N(3-dimethylaminopropyl)-1,3-propylenediamine were used to determine the dielectric properties. Sample of the neat epoxy resin and nanoparticle loaded epoxy resin in the form of disc were prepared of different weight fraction. SiO2 and ZnO nanoparticles were taken as filler in the epoxy resin. Complex permittivity of the prepared samples was measured in the frequency range of 20 Hz to 2 MHz using precision LCR meter at room temperature. The charismatic change in dielectric behavior based on type and concentration of nanoparticle are discussed in detail.

Synthesis of Bi0.9Nd0.1FeO3 multiferroic nanoceramic by sol-gel method and its structural, surface morphology and dielectric analysis

NASA Astrophysics Data System (ADS)

Das, S.; Bera, K. P.; Nath, T. K.

2017-05-01

Synthesis of Nd-doped BFO multiferroic nanoceramic by the conventional sol-gel method has been carried out. HRXRD and FESEM have been used for the structural analysis to confirm the triclinic structure and to obtain the surface morphology showing agglomeration and to find out the size of the nanoparticles. A high precision LCR meter has been used to record the dielectric constants at various temperatures. Several anomalies are observed whose physical explanations have been given. Space charge polarization effect and magnetoelectric coupling have been confirmed in the synthesized Bi0.9Nd0.1FeO3 nanoceramic sample.

Dielectric spectroscopy of solutions of amino silicone emulsion in distilled water

NASA Astrophysics Data System (ADS)

Shah, K. N.; Rana, V. A.; Trivedi, C. M.; Vankar, H. P.

2016-05-01

Complex permittivity spectra ɛ*(ω) = ɛ' - jɛ″ of solutions of amino silicone emulsion in distilled water in the frequency range 100 Hz to 2 MHz were obtained using precision LCR meter. Complex permittivity data is used to find out complex impedance z*(ω) and complex electric conductivity σ*(ω). All these spectra are used to gain information about various polarization processes taking place in the solutions of amino silicone emulsion in distilled water under the effect of ac electric field. The frequency and concentration dependent behavior of the solutions of amino silicone emulsion in distilled waterhave beenalso investigated. Density and refractive index of the samples are also measured and are reported.

Diagnostic accuracy of nucleic acid amplification tests (NAATs) in urine for genitourinary tuberculosis: a systematic review and meta-analysis.

PubMed

Altez-Fernandez, Carlos; Ortiz, Victor; Mirzazadeh, Majid; Zegarra, Luis; Seas, Carlos; Ugarte-Gil, Cesar

2017-06-05

Genitourinary tuberculosis is the third most common form of extrapulmonary tuberculosis. Diagnosis is difficult because of unspecific clinical manifestations and low accuracy of conventional tests. Unfortunately, the delayed diagnosis impacts the urinary tract severely. Nucleic acid amplification tests yield fast results, and among these, new technologies can also detect drug resistance. There is lack of consensus regarding the use of these tests in genitourinary tuberculosis; we therefore aimed to assess the accuracy of nucleic acid amplification tests in the diagnosis of genitourinary tuberculosis and to evaluate the heterogeneity between studies. We did a systematic review and meta-analysis of research articles comparing the accuracy of a reference standard and a nucleic acid amplification test for diagnosis of urinary tract tuberculosis. We searched Medline, EMBASE, Web of Science, LILACS, Cochrane Library, and Scopus for articles published between Jan 1, 1990, and Apr 14, 2016. Two investigators identified eligible articles and extracted data for individual study sites. We analyzed data in groups with the same index test. Then, we generated pooled summary estimates (95% CIs) for sensitivity and specificity by use of random-effects meta-analysis when studies were not heterogeneous. We identified eleven relevant studies from ten articles, giving information on PCR, LCR and Xpert MTB/RIF tests. All PCR studies were "in-house" tests, with different gene targets and had several quality concerns therefore we did not proceed with a pooled analysis. Only one study used LCR. Xpert studies were of good quality and not heterogeneous, pooled sensitivity was 0·87 (0·66-0·96) and specificity was 0·91 (0·84-0·95). PCR studies were highly heterogeneous. Among Xpert MTB/RIF studies, specificity was favorable with an acceptable confidence interval, however new studies can update meta-analysis and get more precise estimates. Further high-quality studies are urgently needed to improve diagnosis of genitourinary tuberculosis. PROSPERO CRD42016039020.

Recurrent reciprocal deletions and duplications of 16p13.11: the deletion is a risk factor for MR/MCA while the duplication may be a rare benign variant

PubMed Central

Hannes, F D; Sharp, A J; Mefford, H C; de Ravel, T; Ruivenkamp, C A; Breuning, M H; Fryns, J-P; Devriendt, K; Van Buggenhout, G; Vogels, A; Stewart, H; Hennekam, R C; Cooper, G M; Regan, R; Knight, S J L; Eichler, E E; Vermeesch, J R

2009-01-01

Background: Genomic disorders are often caused by non-allelic homologous recombination between segmental duplications. Chromosome 16 is especially rich in a chromosome-specific low copy repeat, termed LCR16. Methods and Results: A bacterial artificial chromosome (BAC) array comparative genome hybridisation (CGH) screen of 1027 patients with mental retardation and/or multiple congenital anomalies (MR/MCA) was performed. The BAC array CGH screen identified five patients with deletions and five with apparently reciprocal duplications of 16p13 covering 1.65 Mb, including 15 RefSeq genes. In addition, three atypical rearrangements overlapping or flanking this region were found. Fine mapping by high-resolution oligonucleotide arrays suggests that these deletions and duplications result from non-allelic homologous recombination (NAHR) between distinct LCR16 subunits with >99% sequence identity. Deletions and duplications were either de novo or inherited from unaffected parents. To determine whether these imbalances are associated with the MR/MCA phenotype or whether they might be benign variants, a population of 2014 normal controls was screened. The absence of deletions in the control population showed that 16p13.11 deletions are significantly associated with MR/MCA (p = 0.0048). Despite phenotypic variability, common features were identified: three patients with deletions presented with MR, microcephaly and epilepsy (two of these had also short stature), and two other deletion carriers ascertained prenatally presented with cleft lip and midline defects. In contrast to its previous association with autism, the duplication seems to be a common variant in the population (5/1682, 0.29%). Conclusion: These findings indicate that deletions inherited from clinically normal parents are likely to be causal for the patients’ phenotype whereas the role of duplications (de novo or inherited) in the phenotype remains uncertain. This difference in knowledge regarding the clinical relevance of the deletion and the duplication causes a paradigm shift in (cyto)genetic counselling. PMID:18550696

Potential ecological and human health risks of heavy metals in surface soils associated with iron ore mining in Pahang, Malaysia.

PubMed

Diami, Siti Merryan; Kusin, Faradiella Mohd; Madzin, Zafira

2016-10-01

The composition of heavy metals (and metalloid) in surface soils of iron ore mine-impacted areas has been evaluated of their potential ecological and human health risks. The mining areas included seven selected locations in the vicinity of active and abandoned iron ore-mining sites in Pahang, Malaysia. Heavy metals such as Fe, Mn, Cu, Zn, Co, Pb, Cr, Ni, and Cd and metalloid As were present in the mining soils of the studied area, while Cu was found exceeding the soil guideline value at all sampling locations. However, the assessment of the potential ecological risk index (RI) indicated low ecological risk (RI between 44 and 128) with respect to Cd, Pb, Cu, As, Zn, Co, and Ni in the surface soils. Contributions of potential ecological risk [Formula: see text]by metal elements to the total potential ecological RI were evident for Cd, As, Pb, and Cu. Contribution of Cu appears to be consistently greater in the abandoned mining area compared to active iron ore-mining site. For non-carcinogenic risk, no significant potential health risk was found to both children and adults as the hazard indices (HIs) were all below than 1. The lifetime cancer risk (LCR) indicated that As has greater potential carcinogenic risk compared to other metals that may induce carcinogenic effects such as Pb, Cr, and Cd, while the LCR of As for children fell within tolerable range for regulatory purposes. Irrespective of carcinogenic or non-carcinogenic risk, greater potential health risk was found among children (by an order of magnitude higher for most metals) compared to adults. The hazard quotient (HQ) and cancer risk indicated that the pathways for the risk to occur were found to be in the order of ingestion > dermal > inhalation. Overall, findings showed that some metals and metalloid were still present at comparable concentrations even long after cessation of the iron ore-mining activities.

Genome-Wide Identification of Regulatory Elements and Reconstruction of Gene Regulatory Networks of the Green Alga Chlamydomonas reinhardtii under Carbon Deprivation

PubMed Central

Vischi Winck, Flavia; Arvidsson, Samuel; Riaño-Pachón, Diego Mauricio; Hempel, Sabrina; Koseska, Aneta; Nikoloski, Zoran; Urbina Gomez, David Alejandro; Rupprecht, Jens; Mueller-Roeber, Bernd

2013-01-01

The unicellular green alga Chlamydomonas reinhardtii is a long-established model organism for studies on photosynthesis and carbon metabolism-related physiology. Under conditions of air-level carbon dioxide concentration [CO2], a carbon concentrating mechanism (CCM) is induced to facilitate cellular carbon uptake. CCM increases the availability of carbon dioxide at the site of cellular carbon fixation. To improve our understanding of the transcriptional control of the CCM, we employed FAIRE-seq (formaldehyde-assisted Isolation of Regulatory Elements, followed by deep sequencing) to determine nucleosome-depleted chromatin regions of algal cells subjected to carbon deprivation. Our FAIRE data recapitulated the positions of known regulatory elements in the promoter of the periplasmic carbonic anhydrase (Cah1) gene, which is upregulated during CCM induction, and revealed new candidate regulatory elements at a genome-wide scale. In addition, time series expression patterns of 130 transcription factor (TF) and transcription regulator (TR) genes were obtained for cells cultured under photoautotrophic condition and subjected to a shift from high to low [CO2]. Groups of co-expressed genes were identified and a putative directed gene-regulatory network underlying the CCM was reconstructed from the gene expression data using the recently developed IOTA (inner composition alignment) method. Among the candidate regulatory genes, two members of the MYB-related TF family, Lcr1 (Low-CO 2 response regulator 1) and Lcr2 (Low-CO 2 response regulator 2), may play an important role in down-regulating the expression of a particular set of TF and TR genes in response to low [CO2]. The results obtained provide new insights into the transcriptional control of the CCM and revealed more than 60 new candidate regulatory genes. Deep sequencing of nucleosome-depleted genomic regions indicated the presence of new, previously unknown regulatory elements in the C. reinhardtii genome. Our work can serve as a basis for future functional studies of transcriptional regulator genes and genomic regulatory elements in Chlamydomonas. PMID:24224019

Assessing reproductive and endocrine parameters in male largescale suckers (Catostomus macrocheilus) along a contaminant gradient in the lower Columbia River, USA

USGS Publications Warehouse

Jenkins, Jill A.; Olivier, H.M.; Draugelis-Dale, R. O.; Eilts, B.E.; Torres, L.; Patiño, R.; Nilsen, Elena B.; Goodbred, Steven L.

2014-01-01

Persistent organochlorine pollutants such as polychlorinated biphenyls (PCBs), dichlorodiphenyldichloroethylene (p,p′-DDE), and polybrominated diphenyl ethers (PBDEs) are stable, bioaccumulative, and widely found in the environment, wildlife, and the human population. To explore the hypothesis that reproduction in male fish is associated with environmental exposures in the lower Columbia River (LCR), reproductive and endocrine parameters were studied in male resident, non-anadromous largescale sucker (Catostomus macrocheilus) (LSS) in the same habitats as anadromous salmonids having conservation status. Testes, thyroid tissue and plasma collected in 2010 from Longview (LV), Columbia City (CC), and Skamania (SK; reference) were studied. Sperm morphologies and thyrocyte heights were measured by light microscopy, sperm motilities by computer-assisted sperm motion analysis, sperm adenosine triphosphate (ATP) with luciferase, and plasma vitellogenin (VTG), thyroxine (T4), and triiodothyronine (T3) by immunoassay. Sperm apoptosis, viability, mitochondrial membrane potential, nuclear DNA fragmentation, and reproductive stage were measured by flow cytometry. Sperm quality parameters (except counts) and VTG were significantly different among sites, with correlations between VTG and 7 sperm parameters. Thyrocyte heights, T4, T3, gonadosomatic index and Fulton's condition factor differed among sites, but not significantly. Sperm quality was significantly lower and VTG higher where liver contaminants and water estrogen equivalents were highest (LV site). Total PCBs (specifically PCB-138, -146, -151, -170, -174, -177, -180, -183, -187, -194, and -206) and total PBDEs (specifically BDE-47, -100, -153, and -154) were negatively correlated with sperm motility. PCB-206 and BDE-154 were positively correlated with DNA fragmentation, and pentachloroanisole and VTG were positively correlated with sperm apoptosis and negatively correlated with ATP. BDE-99 was positively correlated with sperm counts and motility; T4 was negatively correlated with counts and positively correlated with motility, thus indicating possible androgenic mechanisms and thyroid endocrine disruption. Male LSS proved to be an informative model for studying reproductive and endocrine biomarkers in the LCR.

Melon chlorotic leaf curl virus: characterization and differential reassortment with closest relatives reveal adaptive virulence in the squash leaf curl virus clade and host shifting by the host-restricted bean calico mosaic virus.

PubMed

Idris, A M; Mills-Lujan, K; Martin, K; Brown, J K

2008-02-01

The genome components of the Melon chlorotic leaf curl virus (MCLCuV) were cloned from symptomatic cantaloupe leaves collected in Guatemala during 2002. The MCLCuV DNA-A and DNA-B components shared their closest nucleotide identities among begomoviruses, at approximately 90 and 81%, respectively, with a papaya isolate of MCLCuV from Costa Rica. The closest relatives at the species level were other members of the Squash leaf curl virus (SLCV) clade, which is endemic in the southwestern United States and Mexico. Biolistic inoculation of cantaloupe seedlings with the MCLCuV DNA-A and -B components resulted in the development of characteristic disease symptoms, providing definitive evidence of causality. MCLCuV experimentally infected species within the Cucurbitaceae, Fabaceae, and Solanaceae. The potential for interspecific reassortment was examined for MCLCuV and its closest relatives, including the bean-restricted Bean calico mosaic virus (BCaMV), and three other cucurbit-infecting species, Cucurbit leaf crumple virus (CuLCrV), SLCV, and SMLCV. The cucurbit viruses have distinct but overlapping host ranges. All possible reassortants were established using heterologous combinations of the DNA-A or DNA-B components. Surprisingly, only certain reassortants arising from MCLCuV and BCaMV, or MCLCuV and CuLCrV, were viable in bean, even though it is a host of all of the "wild-type" (parent) viruses. The bean-restricted BCaMV was differentially assisted in systemically infecting the cucurbit test species by the components of the four cucurbit-adapted begomoviruses. In certain heterologous combinations, the BCaMV DNA-A or -B component was able to infect one or more cucurbit species. Generally, the reassortants were less virulent in the test hosts than the respective wild-type (parent) viruses, strongly implicating adaptive modulation of virulence. This is the first illustration of reassortment resulting in the host range expansion of a host-restricted begomovirus.

Antigenic profiling of Yersinia pestis infection in the Wyoming coyote (Canis latrans)

USGS Publications Warehouse

Vernati, G.; Edwards, W.H.; Rocke, T.E.; Little, S.F.; Andrews, G.P.

2011-01-01

Although Yersinia pestis is classified as a "high-virulence" pathogen, some host species are variably susceptible to disease. Coyotes (Canis latrans) exhibit mild, if any, symptoms during infection, but antibody production occurs postinfection. This immune response has been reported to be against the F1 capsule, although little subsequent characterization has been conducted. To further define the nature of coyote humoral immunity to plague, qualitative serology was conducted to assess the antiplague antibody repertoire. Humoral responses to six plasmid-encoded Y. pestis virulence factors were first examined. Of 20 individual immune coyotes, 90% were reactive to at least one other antigen in the panel other than F1. The frequency of reactivity to low calcium response plasmid (pLcr)-encoded Yersinia protein kinase A (YpkA) and Yersinia outer protein D (YopD) was significantly greater than that previously observed in a murine model for plague. Additionally, both V antigen and plasminogen activator were reactive with over half of the serum samples tested. Reactivity to F1 was markedly less frequent in coyotes (35%). Twenty previously tested antibody-negative samples were also examined. While the majority were negative across the panel, 15% were positive for 1-3 non-F1 antigens. In vivo-induced antigen technology employed to identify novel chromosomal genes of Y. pestis that are up-regulated during infection resulted in the identification of five proteins, including a flagellar component (FliP) that was uniquely reactive with the coyote serum compared with immune serum from two other host species. Collectively, these data suggest that humoral immunity to pLcr-encoded antigens and the pesticin plasmid (pPst)-encoded Pla antigen may be relevant to plague resistance in coyotes. The serologic profile of Y. pestis chromosomal antigens up-regulated in vivo specific to C. latrans may provide insight into the differences in the pathogen-host responses during Y. pestis infection.

Meeting the Deadline: Why, When and How

NASA Technical Reports Server (NTRS)

Dignum, Frank; Broersen, Jan; Dignum, Virginia; Meyer, John-Jules

2004-01-01

A normative system is defined as any set of interacting agents whose behavior can usefully be regarded as norm-directed. Most organizations, and more specifically institutions, fall under this definition. Interactions in these normative systems are regulated by normative templates that describe desired behavior in terms of deontic concepts (obligations, prohibitions and permissions), deadlines, violations and sanctions. Agreements between agents, and between an agent and the society, can then be specified by means of contracts. Contracts provide flexible but verifiable means to integrate society requirements and agent autonomy. and are an adequate means for the explicit specification of interactions. From the society perspective, it is important that these contracts adhere to the specifications described in the model of the organization. If we want to automate such verifications, we have to formalize the languages used for contracts and for the specification of organizations. The logic LCR is based on deontic temporal logic. LCR is an expressive language for describing interaction in multi-agent systems, including obligations with deadlines. Deadlines are important norms in most interactions between agents. Intuitively, a deadline states that an agent should perform an action before a certain point in time. The obligation to perform the action starts at the moment the deadline becomes active. E.g. when a contract is signed or approved. If the action is not performed in time a violation of the deadline occurs. It can be specified independently what measure has to be taken in this case. In this paper we investigate the deadline concept in more detail. The paper is organized as follows. Section 2 defines the variant of CTL we use. In section 3, we discuss the basic intuitions of deadlines. Section 4 presents a first intuitive formalization for deadlines. In section 5, we look at a more complex model for deadlines trying to catch some more practical aspects. Finally, in section 6 we present issues for future work and our conciusions.

Ultrasonographic localization of the thyroid gland for its optimal shielding prior to lateral cephalometric radiography: a pilot study

PubMed Central

Hurmerinta, Kirsti; Rice, David; Suomalainen, Anni

2016-01-01

Objectives: Lateral cephalometric radiography is a common radiographic examination technique in children. The exclusion of the thyroid gland from the primary X-ray beam is important especially with children. However, patient treatment might require displaying the four most cranial cervical vertebrae (C1–C4) for the assessment of cervical vertebral maturation. Our aim was to present a safe way to display C1–C4 and exclude the thyroid gland from the X-ray beam during lateral cephalometric radiography. Methods: The thyroid glands of 25, 7- to 12-year-old patients were localized by ultrasound examination and shielded prior to lateral cephalometric radiography. A roentgen-positive mark was taped on the patient's skin at the level of most cranial level of the thyroid gland in the midsagittal plane. After exposure, each lateral cephalometric radiograph (LCR) was analyzed for the visibility of the cervical vertebrae. The distance between the ear post and the highest edge of the thyroid shield (TS) at the lateral part of the neck was measured and compared with the distance between the centre of the radiological external auditory meatus, and a roentgen-positive mark was made on the LCR. Results: 68% of the LCRs displayed C1–C4, and the rest of them displayed C1–C3. In all of the patients, the highest edge of the TS in the lateral parts of the neck was located in a higher position than the actual most cranial level of the thyroid gland. Conclusions: Despite localizing the thyroid gland prior to lateral cephalometric radiography, simultaneous visualization of C1–C4 and exclusion of the thyroid gland from the primary X-ray beam during lateral cephalometric radiography might not be completely possible in children because of the design and poor fitness of the TS. PMID:26764584

Dielectric relaxation spectroscopy of aqueous solutions of diclofenac potassium over the frequency range of 20 Hz to 2 MHz at 303.15 K temperature

NASA Astrophysics Data System (ADS)

Karakthala, J. B.; Vankar, H. P.; Rana, V. A.

2018-05-01

The complex relative dielectric function ɛ*(ω) = ɛ' - jɛ″ of aqueous solutions of diclofenac potassium (DK) in the frequency range 20 Hz to 2 MHz at 303.15 K was measured using a precision LCR meter. The electrical/dielectric properties of the solutions samples were represented in terms of complex relative dielectric function ɛ*(ω) real part σ'(ω) of complex ac conductivity and dc conductivity. These types of studies can be used to explore various mechanism contributed in the absorption, transportation of drug through tissues and membranes of body as well as interactions of drug with body fluid and blood plasma.

Conductivity, dielectric and modulus study of chitosan-methyl cellulose - BMIMTFSI polymer electrolyte doped with cellulose nano crystal

NASA Astrophysics Data System (ADS)

Misenan, Muhammad Syukri Mohamad; Ali, Ernie Suzana; Khiar, Azwani Sofia Ahmad

2018-06-01

In this study, the effect of adding cellulose nanocrystal (CNC) on the conductivity of biopolymer electrolyte (BPE) based on chitosan-methylcellulose-BMIMTFSI has been studied. The samples were prepared via solution casting technique. The film was characterized by impedance spectroscopy HIOKI 3531- 01 LCR Hi-Tester to measure its ionic conductivity at room temperatures over a wide range of frequency between 50Hz-5MHz. Sample with 15 wt% of CNC shows the highest conductivity of 4.82 x 10-6 Scm-1 at room temperature. Dielectric and modulus studies were carried out to further understands the conductivity behavior of the samples. The increase in conductivity is mainly due to the increase in number of charge carriers.

Microstructural, optical and dielectric properties of La{sub 0.8}Ba{sub 0.2}FeO{sub 3} nanostructures synthesized by sol-gel combustion method

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

Ali, S. Asad, E-mail: asadsyyed@gmail.com; Naseem, Swaleha; Khan, Wasi

2015-06-24

Barium doped lanthanum ferrite (LaFeO{sub 3}) nanoparticles (NPs) were prepared by gel combustion method and calcinated at 700°C. Microstructural studies were carried by XRD and SEM techniques. The results of structural characterization show the formation of all samples in single phase without any impurity. Optical properties were studied by UV- visible technique. The energy band gap was calculated and obtained 3.01 eV. Dielectric properties characterized by LCR meter and have been observed appreciable changes. The observed behavior of the dielectric properties can be attributed on the basis of Koop’s theory based on Maxwell-Wagner two layer models in studied nanoparticles.

Genotoxicity assessment of multispecies probiotics using reverse mutation, mammalian chromosomal aberration, and rodent micronucleus tests.

PubMed

Chiu, Yi-Jen; Nam, Mun-Kit; Tsai, Yueh-Ting; Huang, Chun-Chi; Tsai, Cheng-Chih

2013-01-01

Genotoxicity assessment is carried out on freeze dried powder of cultured probiotics containing Lactobacillus rhamnosus LCR177, Bifidobacterium adolescentis BA286, and Pediococcus acidilactici PA318. Ames tests, in vitro mammalian chromosome aberration assay, and micronucleus tests in mouse peripheral blood are performed. For 5 strains of Salmonella Typhimurium, the Ames tests show no increased reverse mutation upon exposure to the test substance. In CHO cells, the frequency of chromosome aberration does not increase in responding to the treatment of probiotics. Likewise, the frequency of micronucleated reticulocytes in probiotics-fed mice is indistinguishable from that in the negative control group. Taken together, the toxicity assessment studies suggest that the multispecies probiotic mixture does not have mutagenic effects on various organisms.

Voltage Stress on Y Capacitors from Indirect Lightning Pulses According to ED-14/DO-160

NASA Astrophysics Data System (ADS)

Meier, F.

2012-05-01

Transients due to lightning strikes on an aircraft's fuselage impose stress on the input filters of elec- tronic equipment. Permanent damage can occur when exceeding the voltage handling capacity of filter components causing a short circuit to ground. In ED-14/DO-160, section 22, a number of waveforms and levels are defined which are used to check the airworthiness of avionics equipment. Depending on pro- cedure and level, Y-capacitors are stressed by transient voltages which exceed their dielectric strength. The design engineer's task is a properly select the type and voltage rating of capacitors. With moderate simplifications, a LCR-series network is justified to calculate the peak voltage dependent on the capacitance.

Effect of Y2O3 on polyindole for high frequency capacitor application

NASA Astrophysics Data System (ADS)

Maji, P.; Choudhary, R. B.; Majhi, M.

2017-05-01

Polyindole-Yittrium Oxide (PIn-Y2O3) composite was synthesized in the laboratory through chemical polymerization process. The structural and morphological studies of PIn-Y2O3 composite were investigated using X-ray diffraction (XRD) and scanning electron microscopic (SEM) techniques. These studies showed that PIn-Y2O3 composite was amorphous in nature and formed with spherical granule shape. The dielectric response was measured through LCR meter in the frequency range from 100 Hz to 1 MHz. The dielectric studies revealed that incorporation of Y2O3 into polymeric matrix improved the dielectric behavior of PIn polymer and markedly suitable for its application in high frequency capacitor and many other electronic devices.

Evaluation of adhesive-free crossed-electrode poly(vinylidene fluoride) copolymer array transducers for high frequency imaging

NASA Astrophysics Data System (ADS)

Wagle, Sanat; Decharat, Adit; Habib, Anowarul; Ahluwalia, Balpreet S.; Melandsø, Frank

2016-07-01

High frequency crossed-electrode transducers have been investigated, both as single and dual layer transducers. Prototypes of these transducers were developed for 4 crossed lines (yielding 16 square elements) on a polymer substrate, using a layer-by-layer deposition method for poly(vinylidene fluoride-trifluoroethylene) [P(VDF-TrFE)] with intermediate sputtered electrodes. The transducer was characterized using various methods [LCR analyzer, a pulse-echo experimental setup, and a numerical Finite element method (FEM) model] and evaluated in terms of uniformity of bandwidth and acoustical energy output. All 16 transducer elements produced broad-banded ultrasonic spectra with small variation in central frequency and -6 dB bandwidth. The frequency responses obtained experimentally were verified using a numerical model.

Dielectric properties of binary mixtures of methyl iso butyl ketone and amino silicone oil

NASA Astrophysics Data System (ADS)

Shah, K. N.; Rana, V. A.; Trivedi, C. M.; Vankar, H. P.

2017-05-01

Dielectric permittivity ɛ*(ω) = ɛ' - jɛ″ of the binary mixtures of the methyl iso butyl ketone and amino silicone oil in the frequency range 100 Hz to 2 MHz were measured using precision LCR meter at 305.15 K. Relative complex permittivity spectra in the frequency range 100 Hz to 2 MHz, of the mixture solutions of varying concentrations is reported. Determined values of the permittivity at optical frequency of all the samples are also reported. The dielectric parameters are used to gain information about the effect of concentration variation of components of the mixtures on the dielectric properties. It also provides the information about electrode polarization phenomena taking place under the low frequency A.C. electric field.

Frequency dependent dielectric properties of Sr doped NiO nanostructures

NASA Astrophysics Data System (ADS)

Siddique, M. Naseem; Ahmed, Ateeq; Ali, T.; Tripathi, P.

2018-05-01

Ni1-xSrxO (x=0.0, 0.02) nanoparticles have been synthesized using sol-gel method calcined at temperature 600 °C. The XRD analysis result revealed that the calcined sample has a cubic structure with single phase structure. We have calculated crystallite size of samples using both Debye-Sherrer and William Hall (W-H) method which are found to be 19.69 nm, 22.39 nm and 28.50 nm, 33.27 nm, respectively. TEM image reveals the formation of spherical shaped particles. In addition, dielectric properties have been studied using LCR measurement and found that ɛ', ɛ″ and tan δ are decreases with increase in frequency whereas ac conductivity increases with increase in frequency. This behavior may be explained using Maxwell-Wagner and Koop theory.

Dielectric characterization of TiO2, Al2O3 - Nanoparticle loaded epoxy resin

NASA Astrophysics Data System (ADS)

Thakor, S. G.; Rana, V. A.; Vankar, H. P.

2018-05-01

In present work, the dielectric properties of two different nanoparticle loaded Bisphenol A-epoxy resin were carried out at room temperature. Sample of the neat epoxy resin and nanoparticle loaded epoxy resin in the form of disc were prepared of different weight fraction (i.e 0.5 wt%,0.7 wt%,1 wt%,1.5 wt%,1.7 wt%,2 wt%). TiO2 and Al2O3 nanoparticles were taken as filler in the epoxy resin. Complex permittivity of the prepared samples was measured using Agilent E4980A precision LCR meter in frequency range of 103 Hz to 106 Hz. The dependency of dielectric behavior on type and concentration of nanoparticle in considered frequency range are discussed in detail.

Quantitative evaluation method for nonlinear characteristics of piezoelectric transducers under high stress with complex nonlinear elastic constant

NASA Astrophysics Data System (ADS)

Miyake, Susumu; Kasashima, Takashi; Yamazaki, Masato; Okimura, Yasuyuki; Nagata, Hajime; Hosaka, Hiroshi; Morita, Takeshi

2018-07-01

The high power properties of piezoelectric transducers were evaluated considering a complex nonlinear elastic constant. The piezoelectric LCR equivalent circuit with nonlinear circuit parameters was utilized to measure them. The deformed admittance curve of piezoelectric transducers was measured under a high stress and the complex nonlinear elastic constant was calculated by curve fitting. Transducers with various piezoelectric materials, Pb(Zr,Ti)O3, (K,Na)NbO3, and Ba(Zr,Ti)O3–(Ba,Ca)TiO3, were investigated by the proposed method. The measured complex nonlinear elastic constant strongly depends on the linear elastic and piezoelectric constants. This relationship indicates that piezoelectric high power properties can be controlled by modifying the linear elastic and piezoelectric constants.

Selective sensing of vapors of similar dielectric constants using peptide-capped gold nanoparticles on individual multivariable transducers.

PubMed

Nagraj, Nandini; Slocik, Joseph M; Phillips, David M; Kelley-Loughnane, Nancy; Naik, Rajesh R; Potyrailo, Radislav A

2013-08-07

Peptide-capped AYSSGAPPMPPF gold nanoparticles were demonstrated for highly selective chemical vapor sensing using individual multivariable inductor-capacitor-resistor (LCR) resonators. Their multivariable response was achieved by measuring their resonance impedance spectra followed by multivariate spectral analysis. Detection of model toxic vapors and chemical agent simulants, such as acetonitrile, dichloromethane and methyl salicylate, was performed. Dichloromethane (dielectric constant εr = 9.1) and methyl salicylate (εr = 9.0) were discriminated using a single sensor. These sensing materials coupled to multivariable transducers can provide numerous opportunities for tailoring the vapor response selectivity based on the diversity of the amino acid composition of the peptides, and by the modulation of the nature of peptide-nanoparticle interactions through designed combinations of hydrophobic and hydrophilic amino acids.

Dielectric spectroscopy in aqueous solutions of paracetamol over the frequency range of 20 Hz to 2 MHz at 293.15 K temperature

NASA Astrophysics Data System (ADS)

Pandit, T. R.; Rana, V. A.

2018-05-01

Frequency domain dielectric relaxation spectroscopy plays an important role in the study of pharmaceutical drug molecules. The complex relative dielectric permittivity ɛ*(ω) = ɛ' - j ɛ" of aqueous solutions of paracetamol in the frequency range of 20 Hz to 2 MHz at a temperature range of 293.15 K are measured with the help of Agilent precision LCR meter E4980A along with four terminal liquid test fixture Agilent 16452A. Data of complex relative permittivity are used to calculate loss tangent for all concentrations of paracetamol in distilled water. Electrode polarization relaxation time has been calculated for all solutions. Effect of variation of concentrations of paracetamol in distilled water on these dielectric parameters is discussed.

Electrical properties of binary mixtures of amino silicone oil and methyl iso butyl ketone

NASA Astrophysics Data System (ADS)

Shah, K. N.; Rana, V. A.; Vankar, H. P.

2018-05-01

The real and imaginary parts of the dielectric function of the binary mixtures of the methyl iso butyl ketone and amino silicone oil in the frequency range 100 Hz to 2 MHz were measured using precision LCR meter at 305.15 K. The electrical properties such as electrical modulus M*(ω), electrical conductivity σ*(ω) and complex impedance Z*(ω) were calculated using the dielectric function ɛ*(ω). The ionic polarization relaxation time (Τσ) and D.C. conductivity (σdc) were also calculated using electrical properties. The ionic behavior of methyl iso butyl ketone and non-ionic behavior of amino silicone oil are also explained. The electrical parameters are used to gain information about the effect of concentration variation of components of the mixtures on the electrical properties.

Dielectric study of aqueous solutions of sodium dodecyl sulfate in the frequency span 20 Hz to 2 MHz

NASA Astrophysics Data System (ADS)

Kadve, A. M.; Vankar, H. P.; Rana, V. A.

2017-05-01

Dielectric measurements were carried out for aqueous solutions of Sodium Dodecyl Sulfate (SDS) in the frequency span of 20 Hz to 2 MHz at 300.15 K temperature using precision LCR meter. Also the refractive indices were measured for the solutions at 300.15 K temperature using Abbe's refractometer. The measurements were done for ten different concentrations of SDS in distilled water. Determined values of complex permittivity as a function of frequency were used to evaluate other parameters like loss tangent and electric modulus for the liquid samples. The permittivity at optical frequency were also calculated from the measured refractive indices for the aqueous solutions. The effect of concentration variation of SDS in the aqueous solutions on the determined parameters is discussed.

Studies on the growth aspects, structural, thermal, dielectric and third order nonlinear optical properties of solution grown 4-methylpyridinium p-nitrophenolate single crystal

NASA Astrophysics Data System (ADS)

Devi, S. Reena; Kalaiyarasi, S.; Zahid, I. MD.; Kumar, R. Mohan

2016-11-01

An ionic organic optical crystal of 4-methylpyridinium p-nitrophenolate was grown from methanol by slow evaporation method at ambient temperature. Powder and single crystal X-ray diffraction studies revealed the crystal system and its crystalline perfection. The rocking curve recorded from HRXRD study confirmed the crystal quality. FTIR spectral analysis confirmed the functional groups present in the title compound. UV-visible spectral study revealed the optical window and band gap of grown crystal. The thermal, electrical and surface laser damage threshold properties of harvested crystal were examined by using TGA/DTA, LCR/Impedance Analyzer and Nd:YAG laser system respectively. The third order nonlinear optical property of grown crystal was elucidated by Z-scan technique.

A Non-Intrusive Pressure Sensor by Detecting Multiple Longitudinal Waves

PubMed Central

Zhou, Hongliang; Lin, Weibin; Ge, Xiaocheng; Zhou, Jian

2016-01-01

Pressure vessels are widely used in industrial fields, and some of them are safety-critical components in the system—for example, those which contain flammable or explosive material. Therefore, the pressure of these vessels becomes one of the critical measurements for operational management. In the paper, we introduce a new approach to the design of non-intrusive pressure sensors, based on ultrasonic waves. The model of this sensor is built based upon the travel-time change of the critically refracted longitudinal wave (LCR wave) and the reflected longitudinal waves with the pressure. To evaluate the model, experiments are carried out to compare the proposed model with other existing models. The results show that the proposed model can improve the accuracy compared to models based on a single wave. PMID:27527183

Identification of a 3.0-kb Major Recombination Hotspot in Patients with Sotos Syndrome Who Carry a Common 1.9-Mb Microdeletion

PubMed Central

Visser, Remco; Shimokawa, Osamu; Harada, Naoki; Kinoshita, Akira; Ohta, Tohru; Niikawa, Norio; Matsumoto, Naomichi

2005-01-01

Sotos syndrome (SoS) is a congenital dysmorphic disorder characterized by overgrowth in childhood, distinctive craniofacial features, and mental retardation. Haploinsufficiency of the NSD1 gene owing to either intragenic mutations or microdeletions is known to be the major cause of SoS. The common ∼2.2-Mb microdeletion encompasses the whole NSD1 gene and neighboring genes and is flanked by low-copy repeats (LCRs). Here, we report the identification of a 3.0-kb major recombination hotspot within these LCRs, in which we mapped deletion breakpoints in 78.7% (37/47) of patients with SoS who carry the common microdeletion. The deletion size was subsequently refined to 1.9 Mb. Sequencing of breakpoint fragments from all 37 patients revealed junctions between a segment of the proximal LCR (PLCR-B) and the corresponding region of the distal LCR (DLCR-2B). PLCR-B and DLCR-2B are the only directly oriented regions, whereas the remaining regions of the PLCR and DLCR are in inverted orientation. The PLCR, with a size of 394.0 kb, and the DLCR, with a size of of 429.8 kb, showed high overall homology (∼98.5%), with an increased sequence similarity (∼99.4%) within the 3.0-kb breakpoint cluster. Several recombination-associated motifs were identified in the hotspot and/or its vicinity. Interestingly, a 10-fold average increase of a translin motif, as compared with the normal distribution within the LCRs, was recognized. Furthermore, a heterozygous inversion of the interval between the LCRs was detected in all fathers of the children carrying a deletion in the paternally derived chromosome. The functional significance of these findings remains to be elucidated. Segmental duplications of the primate genome play a major role in chromosomal evolution. Evolutionary study showed that the duplication of the SoS LCRs occurred 23.3–47.6 million years ago, before the divergence of Old World monkeys. PMID:15580547

Human Papillomavirus Type 6 and 11 Genetic Variants Found in 71 Oral and Anogenital Epithelial Samples from Australia

PubMed Central

Danielewski, Jennifer A.; Garland, Suzanne M.; McCloskey, Jenny; Hillman, Richard J.; Tabrizi, Sepehr N.

2013-01-01

Genetic variation of 49 human papillomavirus (HPV) 6 and 22 HPV11 isolates from recurrent respiratory papillomatosis (RRP) (n = 17), genital warts (n = 43), anal cancer (n = 6) and cervical neoplasia cells (n = 5), was determined by sequencing the long control region (LCR) and the E6 and E7 genes. Comparative analysis of genetic variability was examined to determine whether different disease states resulting from HPV6 or HPV11 infection cluster into distinct variant groups. Sequence variation analysis of HPV6 revealed that isolates cluster into variants within previously described HPV6 lineages, with the majority (65%) clustering to HPV6 sublineage B1 across the three genomic regions examined. Overall 72 HPV6 and 25 HPV11 single nucleotide variations, insertions and deletions were observed within samples examined. In addition, missense alterations were observed in the E6/E7 genes for 6 HPV6 and 5 HPV11 variants. No nucleotide variations were identified in any isolates at the four E2 binding sites for HPV6 or HPV11, nor were any isolates found to be identical to the HPV6 lineage A or HPV11 sublineage A1 reference genomes. Overall, a high degree of sequence conservation was observed between isolates across each of the regions investigated for both HPV6 and HPV11. Genetic variants identified a slight association with HPV6 and anogenital lesions (p = 0.04). This study provides important information on the genetic diversity of circulating HPV 6 and HPV11 variants within the Australian population and supports the observation that the majority of HPV6 isolates cluster to the HPV6 sublineage B1 with anogenital lesions demonstrating an association with this sublineage (p = 0.02). Comparative analysis of Australian isolates for both HPV6 and HPV11 to those from other geographical regions based on the LCR revealed a high degree of sequence similarity throughout the world, confirming previous observations that there are no geographically specific variants for these HPV types. PMID:23691108

Molecular characterization, tissue tropism, and genetic variability of the novel Mupapillomavirus type HPV204 and phylogenetically related types HPV1 and HPV63

PubMed Central

Šterbenc, Anja; Hošnjak, Lea; Chouhy, Diego; Bolatti, Elisa M.; Oštrbenk, Anja; Seme, Katja; Kocjan, Boštjan J.; Luzar, Boštjan; Giri, Adriana A.; Poljak, Mario

2017-01-01

HPV204 is the only newly identified Mupapillomavirus (Mu-PV) type in more than a decade. To comprehensively characterize HPV204, we performed a detailed molecular analysis of the viral genome and evaluated its clinical relevance in comparison to the other Mu-PVs, HPV1 and HPV63. The 7,227-bp long genome of HPV204 exhibits typical genomic organization of Mu-PVs with eight open reading frames (ORFs) (E6, E7, E1, E2, E8, E4, L2, and L1). We developed three type-specific quantitative real-time PCRs and used them to test a representative collection (n = 1,006) of various HPV-associated benign and malignant neoplasms, as well as samples of clinically normal cutaneous, mucosal, and mucocutaneous origins. HPV204, HPV1, and HPV63 were detected in 1.1%, 2.7%, and 1.9% of samples tested, respectively, and were present in skin and mucosa, suggesting dual tissue tropism of all Mu-PVs. To evaluate the etiological role of Mu-PVs in the development of HPV-associated neoplasms, Mu-PV viral loads per single cell were estimated. HPV1 and HPV63 were present in high viral copy numbers in 3/43 and 1/43 cutaneous warts, respectively, and were identified as the most likely causative agents of these warts. HPV204 viral load was extremely low in a single HPV204-positive cutaneous wart (7.4 × 10−7 viral copies/cell). Hence, etiological association between HPV204 and the development of cutaneous warts could not be established. To the best of our knowledge, this is the first study to evaluate the genetic variability of Mu-PVs by sequencing complete LCR genomic regions of HPV204, HPV1, and HPV63. We detected several nucleotide substitutions and deletions within the LCR genomic regions of Mu-PVs and identified two genetic variants of HPV204 and HPV63 and five genetic variants of HPV1. PMID:28426749

Enhanced dielectric properties of Fe-substituted TiO2 nanoparticles

NASA Astrophysics Data System (ADS)

Ali, T.; Ahmed, Ateeq; Naseem siddique, M.; Tripathi, P.

2018-04-01

We report the structural and dielectric properties Ti1-xFexO2 (0.00 < x < 0.10) nanoparticles (NPs) synthesized by sol-gel method. The synthesized material has been characterized by soft X-ray absorption spectroscopy (SXAS) in order to investigate the fine structure and electronic valence state. SXAS analysis reveals that Fe-ions exist only in 3+ valance state in all the samples. The dielectric properties were studied by the use of LCR impedance spectroscopy. The dielectric constants, dielectric loss and A.C. conductivity have been determined as a function of frequency and composition of iron. At higher frequencies, the materials exhibited high AC Conductivity and low dielectric constant. The above theory could be explained by 'Maxwell Wagner Model' and may provide a new insight to fabricate nanomaterials having possible electrical application.

A MEMS-based Air Flow Sensor with a Free-standing Micro-cantilever Structure.

PubMed

Wang, Yu-Hsiang; Lee, Chia-Yen; Chiang, Che-Ming

2007-10-17

This paper presents a micro-scale air flow sensor based on a free-standingcantilever structure. In the fabrication process, MEMS techniques are used to deposit asilicon nitride layer on a silicon wafer. A platinum layer is deposited on the silicon nitridelayer to form a piezoresistor, and the resulting structure is then etched to create afreestanding micro-cantilever. When an air flow passes over the surface of the cantileverbeam, the beam deflects in the downward direction, resulting in a small variation in theresistance of the piezoelectric layer. The air flow velocity is determined by measuring thechange in resistance using an external LCR meter. The experimental results indicate that theflow sensor has a high sensitivity (0.0284 ω/ms -1 ), a high velocity measurement limit (45ms -1 ) and a rapid response time (0.53 s).

Further development of raccoon poxvirus-vectored vaccines against plague (Yersinia pestis)

USGS Publications Warehouse

Rocke, Tonie E.; Iams, Keith P.; Dawe, S.; Smith, Susan; Williamson, Judy L.; Heisey, Dennis M.; Osorio, Jorge E.

2009-01-01

In previous studies, we demonstrated protection against plague in mice and prairie dogs using a raccoon pox (RCN) virus-vectored vaccine that expressed the F1 capsular antigen of Yersinia pestis. In order to improve vaccine efficacy, we have now constructed additional RCN-plague vaccines containing two different forms of the lcrV (V) gene, including full-length (Vfull) and a truncated form (V307). Mouse challenge studies with Y. pestis strain CO92 showed that vaccination with a combination of RCN-F1 and the truncated V construct (RCN-V307) provided the greatest improvement (P = 0.01) in protection against plague over vaccination with RCN-F1 alone. This effect was mediated primarily by anti-F1 and anti-V antibodies and both contributed independently to increased survival of vaccinated mice.

A flexible dual mode tactile and proximity sensor using carbon microcoils

NASA Astrophysics Data System (ADS)

Han, Hyo Seung; Park, Junwoo; Nguyen, Tien Dat; Kim, Uikyum; Jeong, Soon Cheol; Kang, Doo In; Choi, Hyouk Ryeol

2016-04-01

This paper proposes a flexible dual mode tactile and proximity sensor using Carbon Microcoils (CMCs). The sensor consists of a Flexible Printed Circuit Board (FPCB) electrode layer and a dielectric layer of CMCs composite. In order to avoid damage from frequent contacts, the sensor has all electrodes on the same plane and a polymer covering is placed on the top of the sensor. CMCs can be modeled as complex LCR circuit and the sensitivity of the sensor highly depends on the CMC content. Proper CMC content is experimentally investigated and applied to make the CMCs composite for the dielectric layer. The CMC sensor measures the capacitance for tactile stimulus and inductance for proximity stimulus. A prototype with a size of 30 × 30 × 0.6 𝑚𝑚3, is manufactured and its feasibility is experimentally validated.

Microstructural, optical and electrical properties of LaFe0.5Cr0.5O3 perovskite nanostructures

NASA Astrophysics Data System (ADS)

Ali, S. Asad; Naseem, Swaleha; Khan, Wasi; Sharma, A.; Naqvi, A. H.

2016-05-01

Perovskite nanocrystalline powder of LaFe0.5Cr0.5O3 was synthesized by sol-gel combustion route and characterized by x-ray diffractometer (XRD), scanning electron microscopy (SEM) equipped with EDS, UV-visible and LCR meter at room temperature Rietveld refinement of the XRD data confirms that the sample is in single phase-rhombohedral structure with space group R-3C. SEM micrograph shows clear nanostructure of the sample and EDS ensures the presence of all elements in good stoichiometric. The optical absorption indicates the maximum absorption at 315 nm and optical band gap of 2.94 eV was estimated using Tauc's relation. Dielectric constant (ɛ') and loss were found to decrease with increase in frequencies. The dielectric behavior was explained on the basis of Maxwell-Wagner's two layer model.

Further development of raccoon poxvirus-vectored vaccines against plague (Yersinia pestis).

PubMed

Rocke, Tonie E; Iams, Keith P; Dawe, Sandra; Smith, Susan R; Williamson, Judy L; Heisey, Dennis M; Osorio, Jorge E

2009-12-11

In previous studies, we demonstrated protection against plague in mice and prairie dogs using a raccoon pox (RCN) virus-vectored vaccine that expressed the F1 capsular antigen of Yersinia pestis. In order to improve vaccine efficacy, we have now constructed additional RCN-plague vaccines containing two different forms of the lcrV (V) gene, including full-length (Vfull) and a truncated form (V307). Mouse challenge studies with Y. pestis strain CO92 showed that vaccination with a combination of RCN-F1 and the truncated V construct (RCN-V307) provided the greatest improvement (P=0.01) in protection against plague over vaccination with RCN-F1 alone. This effect was mediated primarily by anti-F1 and anti-V antibodies and both contributed independently to increased survival of vaccinated mice.

Characterization of Residual Medium Peptides from Yersinia pestis Cultures

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

Clowers, Brian H.; Wunschel, David S.; Kreuzer, Helen W.

2013-04-03

Using a range of common microbial medium formulations (TSB, BHI, LB, and G-media), two attenuated strains of Y. pestis (KIM D27 (pgm-) and KIMD1 lcr-) were cultivated in triplicate. These cellular suspensions were used to develop a method of extracting residual medium peptides from the final microbial preparation to assess their relative abundance and identity. Across the conditions examined, which included additional cellular washing and different forms of microbial inactivation, residual medium peptides were detected. Despite the range of growth medium sources used and the associated manufacturing processes used in their production, a high degree of peptide similarity was observedmore » for a given medium recipe. These results demonstrate that residual medium peptides are retained using traditional microbial cultivation techniques and may be used to inform forensic investigations with respect to production deduction.« less

Microstructural, optical and electrical transport properties of Cd-doped SnO2 nanoparticles

NASA Astrophysics Data System (ADS)

Ahmad, Naseem; Khan, Shakeel; Mohsin Nizam Ansari, Mohd

2018-03-01

We have successfully investigated the structural, optical and dielectric properties of Cd assimilated SnO2 nanoparticles synthesized via very convenient precipitation route. The structural properties were studied by x-ray diffraction method (XRD) and Fourier Transform Infrared (FTIR) Spectroscopy. As-synthesized samples in the form of powder were examined for its morphology and average particle size by Transmission electron microscopy (TEM). The optical properties were studied by diffuse reflectance spectroscopy. Dielectric properties such that complex dielectric constant and ac conductivity were investigated by LCR meter. Average crystallite size calculated by XRD and average particle size obtained from TEM were found to be consistent and below 50 nm for all samples. The optical band gap of as-synthesized powder samples from absorption study was found in the range of 3.76 to 3.97 eV. The grain boundary parameters such that Rgb, Cgb and τ were evaluated using impedance spectroscopy.

Structural and transport properties of NdCrO{sub 3} nanoceramics

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

Saha, Sujoy; Sakhya, Anup Pradhan; Sinha, T. P.

2013-02-05

Reitveld refinement of the room temperature powder X-ray diffraction profile of NdCrO{sub 3} (NCO) nanoceramics synthesized by sol-gel processing shows orthorhombic Pnma (D{sub 2h}{sup 16}) space group symmetry. The refined lattice parameters are a = 5.482(3) A, b = 7.689(4) A and c = 5.416(3) A. Transmission electron microscopy (TEM) of NCO shows that the average particle size is around 70 nm. The electrical transport property of NCO is investigated by both conductivity and electric modulus formalism. The electrical data is taken by a LCR meter in a temperature range from 303 K to 573 K and in a frequencymore » range from 42 Hz to 1.1 MHz. The ac conductivity follows a power law. The Cole-Cole plot of impedance at 303 K shows grain effect.« less

A Passive Radio-Frequency Identification (RFID) Gas Sensor With Self-Correction Against Fluctuations of Ambient Temperature

PubMed Central

Potyrailo, Radislav A.; Surman, Cheryl

2013-01-01

Uncontrolled fluctuations of ambient temperature in the field typically greatly reduce accuracy of gas sensors. In this study, we developed an approach for the self-correction against fluctuations of ambient temperature of individual gas and vapor sensors. The main innovation of our work is in the temperature correction which is accomplished without the need for a separate uncoated reference sensor or a separate temperature sensor. Our sensors are resonant inductor-capacitor-resistor (LCR) transducers coated with sensing materials and operated as multivariable passive (battery-free) radio-frequency identification (RFID) sensors. Using our developed approach, we performed quantitation of an exemplary vapor over the temperature range from 25 to 40 °C. This technical solution will be attractive in numerous applications where temperature stabilization of a gas sensor or addition of auxiliary temperature or uncoated reference sensors is prohibitive. PMID:23956496

Structural, optical and dielectric properties of Sn0.97Ce0.03O2 nanostructures

NASA Astrophysics Data System (ADS)

Ahmed, Ateeq; Siddique, M. Naseem; Ali, Tinku; Tripathi, P.

2018-05-01

In present work, 3% cerium doped SnO2 (Sn0.97Ce0.03O2) nanoparticles (NPs) have been synthesized by sol-gel method. The prepared sample has been characterized by using various techniques such as XRD, UV-visible absorption spectroscopy and LCR meter measurements. Structural Rietveld refinement of XRD data reveals that (Sn0.97Ce0.03O2) sample has a pure single phase tetragonal structure with space group (P42/mnm) without creating any impurity phase such as cerium oxide. UV-visible spectroscopy determines band gap value 3.47 eV for (Sn0.97Ce0.03O2) NPs using Tauc's relation. Dielectric constant and loss decreased with increase in frequency while ac conductivity was found to increase with increase in frequency. The observed dielectric results has been explained in the light of Maxwell-Wagner model.

Temperature and induced electric field dependence on the phase transition of 9/70/30, 9/65/35 and 9/60/40 PLZT ceramics

NASA Astrophysics Data System (ADS)

Somwan, Siripong; Funsueb, Narit; Limpichaipanit, Apichart; Ngamjarurojana, Athipong

2018-05-01

In this work, Pb0.91La0.09(Zr1-xTix)0.9775O3 ceramics where x = 0.3, 0.35 and 0.4 (the composition near MPB) were prepared by solid solution method. After fabrication process, electrical property was measured by LCR meter. Polarization and induced strain behavior of the samples were investigated by using interferometry technique modified with Sawyer-Tower circuit at various temperatures. The results of dielectric, polarization and induced strain properties were due to the Zr/Ti ratios, which changed their behavior when temperature was varied (30-70 °C). The normal to macro-micro domains to relaxor and paraelectric phase transition was demonstrated which is related to linear or nonlinear increase of polarization and induced strain as a function of applied subswitching electric field.

A CMOS Self-Contained Quadrature Signal Generator for SoC Impedance Spectroscopy.

PubMed

Márquez, Alejandro; Pérez-Bailón, Jorge; Calvo, Belén; Medrano, Nicolás; Martínez, Pedro A

2018-04-30

This paper presents a low-power fully integrated quadrature signal generator for system-on-chip (SoC) impedance spectroscopy applications. It has been designed in a 0.18 μm-1.8 V CMOS technology as a self-contained oscillator, without the need for an external reference clock. The frequency can be digitally tuned from 10 to 345 kHz with 12-bit accuracy and a relative mean error below 1.7%, thus supporting a wide range of impedance sensing applications. The proposal is experimentally validated in two impedance spectrometry examples, achieving good magnitude and phase recovery results compared to the results obtained using a commercial LCR-meter. Besides the wide frequency tuning range, the proposed programmable oscillator features a total power consumption lower than 0.77 mW and an active area of 0.129 mm², thus constituting a highly suitable choice as stimulation module for instrument-on-a-chip devices.

A MEMS-based Air Flow Sensor with a Free-standing Micro-cantilever Structure

PubMed Central

Wang, Yu-Hsiang; Lee, Chia-Yen; Chiang, Che-Ming

2007-01-01

This paper presents a micro-scale air flow sensor based on a free-standing cantilever structure. In the fabrication process, MEMS techniques are used to deposit a silicon nitride layer on a silicon wafer. A platinum layer is deposited on the silicon nitride layer to form a piezoresistor, and the resulting structure is then etched to create a freestanding micro-cantilever. When an air flow passes over the surface of the cantilever beam, the beam deflects in the downward direction, resulting in a small variation in the resistance of the piezoelectric layer. The air flow velocity is determined by measuring the change in resistance using an external LCR meter. The experimental results indicate that the flow sensor has a high sensitivity (0.0284 Ω/ms-1), a high velocity measurement limit (45 ms-1) and a rapid response time (0.53 s). PMID:28903233

Morphology, Structural and Dielectric Properties of Vacuum Evaporated V2O5 Thin Films

NASA Astrophysics Data System (ADS)

Sengodan, R.; Shekar, B. Chandar; Sathish, S.

Vanadium pentoxide (V2O5) thin films were deposited on well cleaned glass substrate using evaporation technique under the pressure of 10-5 Torr. The thickness of the films was measured by the multiple beam interferometry technique and cross checked by using capacitance method. Metal-Insulator-Metal (MIM) structure was fabricated by using suitable masks to study dielectric properties. The dielectric properties were studied by employing LCR meter in the frequency range 12 Hz to 100 kHz for various temperatures. The temperature co- efficient of permittivity (TCP), temperature co-efficient of capacitance (TCC) and dielectric constant (ɛ) were calculated. The activation energy was calculated and found to be very low. The activation energy was found to be increasing with increase in frequency. The obtained low value of activation energy suggested that the hopping conduction may be due to electrons rather than ions.

Dielectric and electrical properties of binary mixtures of 1-butyl-3-methylimadazolium and water in the frequency range 20 Hz to 2 MHz

NASA Astrophysics Data System (ADS)

Barot, D. K.; Chaube, H. A.; Rana, V. A.

2017-05-01

The complex relative dielectric function ɛ*(ω) = ɛ'-jɛ″ of binary mixture of 1-Butyl-3-methylimadazolium (BMiCl) with water of varying concentration have been measured using Precision LCR meter in the frequency range 20 Hz to 2 MHz at 293.15 K. The dielectric and electrical properties of BMiCl and water are represented in terms of electrical conductivity σ*(ω) and complex relative dielectric function ɛ*(ω). To describe the relationship of the electrical conductivity with concentration, the empirical Casteel-Amis (C-A) equation was used. The influence of concentration variation of BMiCl in water to the various electrical parameters was discussed. All of these presentations are used to explore various processes contributed in the electrical/dielectric properties of the mixtures of BMiCl and water.

Study of complex permittivity spectra of binary mixtures of 2-chloroaniline and methanol in frequency range 10 KHz to 2 MHz at different temperatures

NASA Astrophysics Data System (ADS)

Shah, N. S.; Vankar, H. P.; Rana, V. A.

2017-05-01

The complex relative dielectric function ɛ*(ω)=ɛ'-jɛ″ of the binary mixture of 2-chloroaniline(2-CA) and methanol (MeOH) were measured using precision LCR meter in the frequency range of 10 KHz to 2 MHz The measurements were carried out at eight different temperatures and five different concentrations of 2-CA and MeOH. The loss tangent peaks were observed in the studied frequency range for all the binary mixtures. From the loss tangent peaks electrode polarization relaxation time were evaluated. In the plot of real part of complex permittivity against frequency, at different temperatures for 2-CA (54.54%) + MeOH (45.45%) and 2-CA (27.27%) + MeOH (72.72%)and 100% MeOH systems permittivity inversion effect was observed.

Studies on structural and electrical properties of nanostructured RMnO3 (R = Gd & Ho)

NASA Astrophysics Data System (ADS)

Sapana, Solanki; Dhruv, Davit; Joshi, Zalak; Gadani, Keval; Rathod, K. N.; Boricha, Hetal; Shrimali, V. G.; Trivedi, R. K.; Joshi, A. D.; Pandya, D. D.; Solanki, P. S.; Shah, N. A.

2017-05-01

We report the results of the studies on the structural and electrical properties of multiferroic GdMnO3 and HoMnO3 materials synthesized by sol-gel route. Structural analysis of the results of X-ray diffraction (XRD) measurement shows that materials are found to be crystallized in orthorhombic and hexagonal symmetry, respectively for GdMnO3 and HoMnO3. Frequency dependent dielectric properties of nanostructured GdMnO3 and HoMnO3 were carried out using LCR meter in the frequency range of 100Hz to 2MHz at room temperature. Dielectric constant decreases with increasing frequency for both the nanostructured multiferroics which can be attributed to the dipole relaxation process. AC conductivity (σAC) has been measured for both the samples and fitted theoretically by using power law equation.

Sequence Complexity of Amyloidogenic Regions in Intrinsically Disordered Human Proteins

PubMed Central

Das, Swagata; Pal, Uttam; Das, Supriya; Bagga, Khyati; Roy, Anupam; Mrigwani, Arpita; Maiti, Nakul C.

2014-01-01

An amyloidogenic region (AR) in a protein sequence plays a significant role in protein aggregation and amyloid formation. We have investigated the sequence complexity of AR that is present in intrinsically disordered human proteins. More than 80% human proteins in the disordered protein databases (DisProt+IDEAL) contained one or more ARs. With decrease of protein disorder, AR content in the protein sequence was decreased. A probability density distribution analysis and discrete analysis of AR sequences showed that ∼8% residue in a protein sequence was in AR and the region was in average 8 residues long. The residues in the AR were high in sequence complexity and it seldom overlapped with low complexity regions (LCR), which was largely abundant in disorder proteins. The sequences in the AR showed mixed conformational adaptability towards α-helix, β-sheet/strand and coil conformations. PMID:24594841

Microstructural, Optical and Dielectric Properties of Al-Incorporated SnO2 Nanoparticles

NASA Astrophysics Data System (ADS)

Ahmed, Ateeq; Tripathi, P.; Naseem Siddique, M.; Ali, Tinku

2017-08-01

In this work, Pure SnO2 and Al doped SnO2 nanoparticles with the composition Sn1-xAlxO2 (x = 0, and 0.05) have been successfully prepared using sol-gel technique. The effect of Al dopant on microstructural, optical and dielectric properties has been investigated by X-ray diffraction (XRD), Scanning electron microscopy (SEM), Ultraviolet (UV-Visible) absorption spectroscopy andImpedance spectroscopy (LCR meter)respectively. The XRD patterns indicated tetragonal rutile structure with single phase without any detectable impurity for all samples and incorporation of Al ions into the SnO2 lattice. Crystalline size decreased with aluminum content. The results of SEM confirm nanoparticles size decreases with Al dopant. UV-Visible results showed that optical band also decreases when Al is doped into pure SnO2 lattice. Frequency dependent dielectric properties of pure and doped SnO2 nanoparticles have been also studied.

A facile growth mechanism, structural, optical, dielectric and electrical properties of ZnSe nanosphere via hydrothermal process

NASA Astrophysics Data System (ADS)

Javed, Qurat-Ul-Ain; Baqi, Sabah; Abbas, Hussain; Bibi, Maryam

2017-02-01

Hydrothermal method was chosen as a convenient method to fabricate zinc selenide (ZnSe) nanoparticle materials. The prepared nanospheres were characterized using X-ray diffraction (XRD) and scanning electron microscopy (SEM), where its different properties were observed using UV-visible spectroscopy and LCR meter. It was found that the pure ZnSe nanoparticles have a Zinc blende structure with crystallite size 10.91 nm and in a spherical form with average diameter of 35 nm (before sonication) and 18 nm (after sonication) with wide band gap of 4.28 eV. It was observed that there is inverse relation of frequency with dielectric constant and dielectric loss while AC conductivity grows up by increasing frequency. Such nanostructures were determined to be effectively used in optoelectronic devices as UV detector and in those devices where high-dielectric constant materials are required.

Application of Terahertz Field Enhancement Effect in Metal Microstructures

NASA Astrophysics Data System (ADS)

Nakajima, M.; Kurihara, T.; Tadokoro, Y.; Kang, B.; Takano, K.; Yamaguchi, K.; Watanabe, H.; Oto, K.; Suemoto, T.; Hangyo, M.

2016-12-01

Applications of high-field terahertz pulses are attractive in physics and terahertz technology. In this study, two applications related to high-intensity terahertz pulses are demonstrated. The field enhancement effect by subwavelength metallic microstructures is utilized for terahertz excitation measurement. The spin precession dynamics in magnetic materials was induced by a terahertz magnetic field. Spin precession was amplified by one order of magnitude in amplitude by the enhanced magnetic terahertz field in orthoferrite ErFeO3 with metal microstructures. The induced spin dynamics was analyzed and explained by LLG-LCR model. Moreover, a detection method for terahertz pulses was developed using a cholesteric liquid crystal at room temperature without any electronic devices. The beam profile of terahertz pulses was visualized and compared to other methods such as the knife edge method using pyroelectric detector and micro-bolometer array. The liquid crystal terahertz imager is very simple and has good applicability as a portable terahertz-sensing card.

Electrical description of N2 capacitively coupled plasmas with the global model

NASA Astrophysics Data System (ADS)

Cao, Ming-Lu; Lu, Yi-Jia; Cheng, Jia; Ji, Lin-Hong; Engineering Design Team

2016-10-01

N2 discharges in a commercial capacitively coupled plasma reactor are modelled by a combination of an equivalent circuit and the global model, for a range of gas pressure at 1 4 Torr. The ohmic and inductive plasma bulk and the capacitive sheath are represented as LCR elements, with electrical characteristics determined by plasma parameters. The electron density and electron temperature are obtained from the global model in which a Maxwellian electron distribution is assumed. Voltages and currents are recorded by a VI probe installed after the match network. Using the measured voltage as an input, the current flowing through the discharge volume is calculated from the electrical model and shows excellent agreement with the measurements. The experimentally verified electrical model provides a simple and accurate description for the relationship between the external electrical parameters and the plasma properties, which can serve as a guideline for process window planning in industrial applications.

Deletion of the X-linked opsin gene array locus control region (LCR) results in disruption of the cone mosaic.

PubMed

Carroll, Joseph; Rossi, Ethan A; Porter, Jason; Neitz, Jay; Roorda, Austin; Williams, David R; Neitz, Maureen

2010-09-15

Blue cone monochromacy (BCM) is an X-linked condition in which long- (L) and middle- (M) wavelength-sensitive cone function is absent. Due to the X-linked nature of the condition, female carriers are spared from a full manifestation of the associated defects but can show visual symptoms, including abnormal cone electroretinograms. Here we imaged the cone mosaic in four females carrying an L/M array with deletion of the locus control region, resulting in an absence of L/M opsin gene expression (effectively acting as a cone opsin knockout). On average, they had cone mosaics with reduced density and disrupted organization compared to normal trichromats. This suggests that the absence of opsin in a subset of cones results in their early degeneration, with X-inactivation the likely mechanism underlying phenotypic variability in BCM carriers. Copyright 2010 Elsevier Ltd. All rights reserved.

Synthesis, characterization and AC conductivity studies of silver doped conducting polyaniline/graphene/SrTiO3 composites

NASA Astrophysics Data System (ADS)

Vinay, K.; Shivakumar, K.; Ravikiran, Y. T.; Revanasiddappa, M.

2018-05-01

The present work is an investigation of ac conduction behaviour and dielectric response of Polyaniline/Ag/Graphene/SrTiO3 (PAGS) composite prepared by in-situ chemical oxidative interfacial polymerization using (NH4)2S2O8 as an oxidising agent at 0-5°C. The structural characterization of the samples was examined using FT-IR and XRD techniques. The ac conductivity and dielectric response of synthesized polymer composites were investigated at room temperature in the frequency range varying from 5 × 101 - 5 × 106 Hz using HIOKI make 3532-50 LCR Hi-tester. The ac conductivity increases with increase in frequency and follows the regular trend, the real dielectric constant (ɛ') and imaginary dielectric constant (ɛ'') decreases with increase in frequency and exhibits almost zero dielectric loss at higher frequencies, which suggests that the composite is a lossless material at frequencies beyond 3Hz.

Characterization of Portuguese Centenarian Eating Habits, Nutritional Biomarkers, and Cardiovascular Risk: A Case Control Study

PubMed Central

Valente, A.; Chaves, C.; Gil, A.; Santos, A. C.; Gorjão-Clara, J. P.; Bicho, M.

2018-01-01

Background and Aims Eating habits may contribute to longevity. We characterized the eating habits and cardiovascular risk (CVR) biomarkers in Portuguese centenarians (CENT) compared to controls. Methods and Results Centenarians (n = 253), 100.26 ± 1.98 years, were compared with 268 controls (67.51 ± 3.25), low (LCR) and high (HCR) CVR (QRISK®2-2016). Anthropometric and body composition were evaluated by bioimpedance. Abdominal obesity, BMI, and fat mass (FM) cut-offs were according to the WHO. Sarcopenia was defined by muscle mass index cut-off ≤ 16.7 kg/m2. Daily red meat intake, adjusted for age and gender, was sarcopenia protective (OR = 0.25, 95% CI = 0.096–0.670, P = 0.006); however, it contributes for FM excess (OR = 4.946, 95% CI = 1.471–16.626, P = 0.01), overweight, and obesity (OR = 4.804, 95% CI = 1.666–13.851, P = 0.004). This centenarian eating habit (2%) contrasts to HCR (64.3%). The history of red meat (P < 0.0001) and canned/industrialized food intakes (P < 0.0001) was associated with HCR. Basal metabolism was lower in centenarians versus LCR/HCR (CENT = 1176.78 ± 201.98; LCR = 1356.54 ± 170.65; HCR = 1561.33 ± 267.85; P < 0.0001), BMI (CENT = 21.06 ± 3.68; LCR = 28.49 ± 4.69; HCR = 29.56 ± 5.26; P < 0.0001), waist circumference (CENT = 85.29 ± 10.83; LCR = 96.02 ± 11.71; HCR = 104.50 ± 11.84; P < 0.0001), and waist-hip ratio (CENT = 0.88 ± 0.07; LCR = 0.92 ± 0.08; HCR = 1.01 ± 0.08; P < 0.0001). CENT had lower total cholesterol, LDL cholesterol, non-HDL cholesterol, and cholesterol/HDL ratio than controls. Conclusions Frequent consumption of red meat, cholesterol, and heme iron rich may contribute to obesity and increased CVR. The low frequency of this consumption, observed in centenarians, although associated with sarcopenia, may be one of the keys to longevity. PMID:29725498

Characterization of Portuguese Centenarian Eating Habits, Nutritional Biomarkers, and Cardiovascular Risk: A Case Control Study.

PubMed

da Silva, A Pereira; Valente, A; Chaves, C; Matos, A; Gil, A; Santos, A C; Gorjão-Clara, J P; Bicho, M

2018-01-01

Eating habits may contribute to longevity. We characterized the eating habits and cardiovascular risk (CVR) biomarkers in Portuguese centenarians (CENT) compared to controls. Centenarians ( n = 253), 100.26 ± 1.98 years, were compared with 268 controls (67.51 ± 3.25), low (LCR) and high (HCR) CVR (QRISK®2-2016). Anthropometric and body composition were evaluated by bioimpedance. Abdominal obesity, BMI, and fat mass (FM) cut-offs were according to the WHO. Sarcopenia was defined by muscle mass index cut-off ≤ 16.7 kg/m 2 . Daily red meat intake, adjusted for age and gender, was sarcopenia protective (OR = 0.25, 95% CI = 0.096-0.670, P = 0.006); however, it contributes for FM excess (OR = 4.946, 95% CI = 1.471-16.626, P = 0.01), overweight, and obesity (OR = 4.804, 95% CI = 1.666-13.851, P = 0.004). This centenarian eating habit (2%) contrasts to HCR (64.3%). The history of red meat ( P < 0.0001) and canned/industrialized food intakes ( P < 0.0001) was associated with HCR. Basal metabolism was lower in centenarians versus LCR/HCR (CENT = 1176.78 ± 201.98; LCR = 1356.54 ± 170.65; HCR = 1561.33 ± 267.85; P < 0.0001), BMI (CENT = 21.06 ± 3.68; LCR = 28.49 ± 4.69; HCR = 29.56 ± 5.26; P < 0.0001), waist circumference (CENT = 85.29 ± 10.83; LCR = 96.02 ± 11.71; HCR = 104.50 ± 11.84; P < 0.0001), and waist-hip ratio (CENT = 0.88 ± 0.07; LCR = 0.92 ± 0.08; HCR = 1.01 ± 0.08; P < 0.0001). CENT had lower total cholesterol, LDL cholesterol, non-HDL cholesterol, and cholesterol/HDL ratio than controls. Frequent consumption of red meat, cholesterol, and heme iron rich may contribute to obesity and increased CVR. The low frequency of this consumption, observed in centenarians, although associated with sarcopenia, may be one of the keys to longevity.

Particulate pollutants in the Brazilian city of São Paulo: 1-year investigation for the chemical composition and source apportionment

NASA Astrophysics Data System (ADS)

Martins Pereira, Guilherme; Teinilä, Kimmo; Custódio, Danilo; Gomes Santos, Aldenor; Xian, Huang; Hillamo, Risto; Alves, Célia A.; Bittencourt de Andrade, Jailson; Olímpio da Rocha, Gisele; Kumar, Prashant; Balasubramanian, Rajasekhar; de Fátima Andrade, Maria; de Castro Vasconcellos, Pérola

2017-10-01

São Paulo in Brazil has relatively relaxed regulations for ambient air pollution standards and often experiences high air pollution levels due to emissions of particulate pollutants from local sources and long-range transport of air masses impacted by biomass burning. In order to evaluate the sources of particulate air pollution and related health risks, a year-round sampling was done at the University of São Paulo campus (20 m a.g.l.), a green area near an important expressway. The sampling was performed for PM2. 5 ( ≤ 2. 5 µm) and PM10 ( ≤ 10 µm) in 2014 through intensive (everyday sampling in wintertime) and extensive campaigns (once a week for the whole year) with 24 h of sampling. This year was characterized by having lower average precipitation compared to meteorological data, and high-pollution episodes were observed all year round, with a significant increase in pollution level in the intensive campaign, which was performed during wintertime. Different chemical constituents, such as carbonaceous species, polycyclic aromatic hydrocarbons (PAHs) and derivatives, water-soluble ions, and biomass burning tracers were identified in order to evaluate health risks and to apportion sources. The species such as PAHs, inorganic and organic ions, and monosaccharides were determined using chromatographic techniques and carbonaceous species using thermal-optical analysis. Trace elements were determined using inductively coupled plasma mass spectrometry. The risks associated with particulate matter exposure based on PAH concentrations were also assessed, along with indexes such as the benzo[a]pyrene equivalent (BaPE) and lung cancer risk (LCR). High BaPE and LCR were observed in most of the samples, rising to critical values in the wintertime. Also, biomass burning tracers and PAHs were higher in this season, while secondarily formed ions presented low variation throughout the year. Meanwhile, vehicular tracer species were also higher in the intensive campaign, suggesting the influence of lower dispersion conditions in that period. Source apportionment was performed using positive matrix factorization (PMF), which indicated five different factors: road dust, industrial emissions, vehicular exhaust, biomass burning and secondary processes. The results highlighted the contribution of vehicular emissions and the significant input from biomass combustion in wintertime, suggesting that most of the particulate matter is due to local sources, in addition to the influence of pre-harvest sugarcane burning.

Advanced Materials for Space Applications

NASA Technical Reports Server (NTRS)

Pater, Ruth H.; Curto, Paul A.

2005-01-01

Since NASA was created in 1958, over 6400 patents have been issued to the agency--nearly one in a thousand of all patents ever issued in the United States. A large number of these inventions have focused on new materials that have made space travel and exploration of the moon, Mars, and the outer planets possible. In the last few years, the materials developed by NASA Langley Research Center embody breakthroughs in performance and properties that will enable great achievements in space. The examples discussed below offer significant advantages for use in small satellites, i.e., those with payloads under a metric ton. These include patented products such as LaRC SI, LaRC RP 46, LaRC RP 50, PETI-5, TEEK, PETI-330, LaRC CP, TOR-LM and LaRC LCR (patent pending). These and other new advances in nanotechnology engineering, self-assembling nanostructures and multifunctional aerospace materials are presented and discussed below, and applications with significant technological and commercial advantages are proposed.

Bubbling route to strange nonchaotic attractor in a nonlinear series LCR circuit with a nonsinusoidal force.

PubMed

Senthilkumar, D V; Srinivasan, K; Thamilmaran, K; Lakshmanan, M

2008-12-01

We identify an unconventional route to the creation of a strange nonchaotic attractor (SNA) in a quasiperiodically forced electronic circuit with a nonsinusoidal (square wave) force as one of the quasiperiodic forces through numerical and experimental studies. We find that bubbles appear in the strands of the quasiperiodic attractor due to the instability induced by the additional square-wave-type force. The bubbles then enlarge and get increasingly wrinkled as a function of the control parameter. Finally, the bubbles get extremely wrinkled (while the remaining parts of the strands of the torus remain largely unaffected) resulting in the creation of the SNA; we term this the bubbling route to the SNA. We characterize and confirm this creation from both experimental and numerical data using maximal Lyapunov exponents and their variance, Poincaré maps, Fourier amplitude spectra, and spectral distribution functions. We also strongly confirm the creation of a SNA via the bubbling route by the distribution of the finite-time Lyapunov exponents.

Fabrication of poly(o-anisidine) coated silica core-shell microspheres and their electrorheological response

NASA Astrophysics Data System (ADS)

Lee, Chul Joo; Choi, Hyoung Jin

2017-11-01

In this work, silica/poly(o-anisidine) (POA) core-shell structured microspheres were synthesized by coating the silica core surface with POA with a help of a chemical grafting agent, N-[(3-trimethoxylsilyl)-propyl] aniline. The synthesized silica microspheres were then applied as a polymer/inorganic composite particle-based electrorheological (ER) fluid. The morphology of the silica/POA microspheres was examined by using both transmission electron microscopy and scanning electron microscopy, while their thermal properties and chemical structure were checked by thermogravimetric analysis and Fourier-transform infrared spectroscopy, respectively. The ER properties of the silica/POA particle-based ER fluid were examined by using a Couette-type rotational rheometer equipped with a high-voltage power supplier and analyzed by the Bingham model and modified Mason number. In order to obtain additional information about the electrical polarization properties, the dielectric spectra were measured by an LCR meter and fitted by using the Cole-Cole equation. Furthermore, suspension stability of the ER fluid was tested using Turbiscan.

Preparation of immobilized glucose oxidase wafer enzyme on calcium-bentonite modified by surfactant

NASA Astrophysics Data System (ADS)

Widi, R. K.; Trisulo, D. C.; Budhyantoro, A.; Chrisnasari, R.

2017-07-01

Wafer glucose oxidase (GOx) enzymes was produced by addition of PAH (Poly-Allyamine Hydrochloride) polymer into immobilized GOx enzyme on modified-Tetramethylammonium Hydroxide (TMAH) 5%-calsium-bentonite. The use of surfactant molecul (TMAH) is to modify the surface properties and pore size distribution of the Ca-bentonite. These properties are very important to ensure GOx molecules can be bound on the Ca-bentonit surface to be immobilized. The addition of the polymer (PAH) is expected to lead the substrates to be adsorbed onto the enzyme. In this study, wafer enzymes were made in various concentration ratio (Ca-bentonite : PAH) which are 1:0, 1:1, 1:2 and 1:3. The effect of PAH (Poly-Allyamine Hydrochloride) polymer added with various ratios of concentrations can be shown from the capacitance value on LCR meter and enzyme activity using DNS method. The addition of the polymer (PAH) showed effect on the activity of GOx, it can be shown from the decreasing of capacitance value by increasing of PAH concentration.

Microwave assisted synthesis and characterization of barium titanate nanoparticles for multi layered ceramic capacitor applications.

PubMed

Thirumalai, Sundararajan; Shanmugavel, Balasivanandha Prabu

2011-01-01

Barium titanate is a common ferroelectric electro-ceramic material having high dielectric constant, with photorefractive effect and piezoelectric properties. In this research work, nano-scale barium titanate powders were synthesized by microwave assisted mechano-chemical route. Suitable precursors were ball milled for 20 hours. TGA studies were performed to study the thermal stability of the powders. The powders were characterized by XRD, SEM and EDX Analysis. Microwave and Conventional heating were performed at 1000 degrees C. The overall heating schedule was reduced by 8 hours in microwave heating thereby reducing the energy and time requirement. The nano-scale, impurity-free and defect-free microstructure was clearly evident from the SEM micrograph and EDX patterns. LCR meter was used to measure the dielectric constant and dielectric loss values at various frequencies. Microwave heated powders showed superior dielectric constant value with low dielectric loss which is highly essential for the fabrication of Multi Layered Ceramic Capacitors.

Health service staff's attitudes towards patients with mental illness.

PubMed

Arvaniti, Aikaterini; Samakouri, Maria; Kalamara, Eleni; Bochtsou, Valentini; Bikos, Constantinos; Livaditis, Miltos

2009-08-01

Stereotypes and prejudices against patients with mental illness are widespread in many societies. The aim of the present study is to investigate such attitudes among the staff and medical students, including employees and trainees, in a general university hospital. Six hundred individuals (361 employees, 231 students) completed the following questionnaires: Level of Contact Report (LCR), Authoritarianism Scale (AS), and Opinion about Mental Illness (OMI), a scale yielding five factors (social discrimination, social restriction, social care, social integration, and aetiology). Multivariate linear regression models were applied in order to search for the simultaneous effect of many variables on the scores of OMI factors. An important part of the sample held negative attitudes especially concerning social discrimination and restriction of the patients. Women, older and less educated staff, nursing staff, and people scoring higher on authoritarianism were more prejudiced. Higher scores on familiarity were associated with less negative attitudes. The results indicate the need to develop sensitisation and training programs considering mental health topics among health service employees.

2015 WFNDEC eddy current benchmark modeling of impedance variation in coil due to a crack located at the plate edge

NASA Astrophysics Data System (ADS)

Rocha, João Vicente; Camerini, Cesar; Pereira, Gabriela

2016-02-01

The 2015 World Federation of NDE Centers (WFNDEC) eddy current benchmark problem involves the inspection of two EDM notches placed at the edge of a conducting plate with a pancake coil that runs parallel to the plate's edge line. Experimental data consists of impedance variation measured with a precision LCR bridge as a XY scanner moves the coil. The authors are pleased to present the numerical results obtained with commercial FEM packages (OPERA 3-D). Values of electrical resistance and inductive reactance variation between base material and the region around the notch are plotted as function of the coil displacement over the plate. The calculations were made for frequencies of 1 kHz and 10 kHz and agreement between experimental and numerical results are excellent for all inspection conditions. Explanations are made about how the impedance is calculated as well as pros and cons of the presented methods.

Fabrication of PbFe12O19 nanoparticles and study of their structural, magnetic and dielectric properties

NASA Astrophysics Data System (ADS)

Mousavi Ghahfarokhi, S. E.; Rostami, Z. A.; Kazeminezhad, I.

2016-02-01

In this study, M-type Lead hexaferrite (PbFe12O19) nanoparticles were prepared by a sol-gel method and the prepared powders were annealed at 700-1000 °C for 1, 1.5, 2, 2.5 and 3 h. The Lead hexaferrite powders were characterized using thermogravimetry-differential thermal analysis, X-ray diffraction, scanning electron microscopy, LCR meter, vibrating sample magnetometer, and Fourier transforms infrared spectroscopy. The size of the nanoparticles was increased with the annealing temparature. The results reveal that the best annealing temperature and annealing time for preparing PbFe12O19 nanoparticles at 800 °C and 3 h are obtained. The infrared spectra measured in range of 4000-400 cm-1 exhibit stretching modes of metal ions in tetrahedral site at 580-550 cm-1 and octahedral site at 470-430 cm-1. The variation in ac conductivity (σac) with frequency shows that the electrical conductivity in these ferrites is mainly attributed to the electron hopping mechanism.

Fabrication and analysis of radiofrequency MEMS series capacitive single-pole double-throw switch

NASA Astrophysics Data System (ADS)

Bansal, Deepak; Bajpai, Anuroop; Kumar, Prem; Kaur, Maninder; Rangra, Kamaljit

2016-10-01

A compact radiofrequency (RF) MEMS single-pole double-throw (SPDT) switch based on series capacitive configuration is proposed. The critical process parameters are analyzed to improve the fabrication process. A technique of cold-hot thermal shock for lift-off method is explored. The residual stress in the structure is quantified by lancet test structures that come out to be 51 MPa. Effect of residual stress on actuation voltage is explored, which changes its value from 24 to 22 V. Resonance frequency and switching speed of the switch are 11 kHz and 44 μs, respectively, measured using laser Doppler vibrometer. Measured bandwidth of the SPDT switch is 20 GHz (5 to 25 GHz), which is verified with finite element method simulations in high frequency structure simulator©; and an equivalent LCR circuit in advanced design system©;. Insertion loss of the switch lies in -0.1 to -0.5 dB with isolation better than -20 dB for the above-mentioned bandwidth.