Urgent operation improves weakness in cauda equina syndrome due to lumbar disc herniation.
This study aims to examine the effect of surgical timing on the sphincter function and improvement of motor function in patients with cauda equine syndrome (CES) due to lumbar disc herniation (LDH). Between January 2005 and December 2013, a total of 33 patients (18 males, 15 females; mean age 48.6±2.2 years, range, 24 to 73 years) who underwent lumbar spinal surgery and were diagnosed with CES due to LDH were retrospectively analyzed. Data including demographics, muscle weakness, sensory deficit, sphincter control, LDH level, time from the initiation of symptoms to admission, and time to surgery were documented. The latest muscle weakness, sensory deficits, and sphincter control were also recorded. The patients were divided into two groups according to the rate of muscle strength improvement and data including age, sex, and operation time were compared. The weakness remained unchanged in 11 (33.3%), improved in 13 (39.4%), and returned to normal in nine (27.3%) patients. Sphincter control resolved in five patients. Sensory loss resolved in two patients. While admission duration was shorter in the group with improved muscle strength (p=0.02), there was no significant difference in the time to surgery (p=0.63). Logistic regression analyses revealed that only the admission within 0 to 24 hours was significant for the muscle strength improvement (regression coeeficient [B]=2.83, standard error [SE]=0.86, p=0.006). A significant improvement in the motor strength can be achieved in patients with CES who are operated within 24 hours. On the other hand, patients with CES should be received surgery immediately when first seen, regardless of the time, since the muscle strength is slightly improved.


Contralateral projections of barosensitive neurons of the nucleus tractus solitarii.
Axonal projections of single barosensitive neurons of the nucleus tractus solitarii (NTS) which fired in synchrony with heartbeat, were studied in Nembutal-anesthetized, paralyzed, and artificially ventilated cats. The majority of them were orthodromically activated by electrical stimulation of the ipsilateral cervical vagal nerve. Antidromic mapping by electrical stimulation of the medulla could identify the axonal projections in 14 of the 25 barosensitive NTS neurons examined. Their stem axons crossed the midline to the contralateral side and ascended rostrally. The contralateral axons of some neurons issued collaterals in the rostral ventrolateral medulla (RVLM) medial to and ventromedial to the facial nucleus. The contralateral axons of the other neurons ascended toward the pons without any sign of issuing collaterals within the medulla.


Using technology to promote self-efficacy for healthy eating in adolescents.
To test the effects of a classroom and World Wide Web (WWW) educational intervention on self-efficacy (SE) for healthy eating (HE) and to examine the relationship of the theoretical concepts in a hypothesized model of eating behavior in adolescents. A quasiexperimental pretest, posttest design was used in a random sample of students in two junior high schools. The intervention consisted of a combination of 5 hours of Web-based instruction and 10 hours of classroom curriculum, compared to nutrition education embedded in the standard school curriculum during a 1-month period. Participants completed six questionnaires. Data were analyzed using descriptive statistics, t tests, and Pearson's r correlation coefficient. The intervention group had significantly higher scores for SE for fruits and vegetables, SE for lower fat, usual food choices, and dietary knowledge of fat compared to the control group. No difference was found between groups in food consumption. Self-efficacy was significantly associated with dietary knowledge of lower fat, usual food choices, and was inversely associated with lower-fat consumption in the hypothesized model of eating behavior. The intervention was tailored to the social and developmental preferences of adolescents and effectively increased SE for HE. The mediating role of SE in the hypothesized model of adolescent eating behavior warrants further investigation.


Distinguishing deliberate from systematic thinking.
Deliberate thinking and systematic thinking are often conflated when contrasted with intuitive thinking. We argue that, in fact, nonintuitive thinking is multidimensional, and that deliberate and systematic thinking are distinct nonintuitive processes. We establish their distinct meanings in 6 studies using 3 research paradigms. Our first paradigm (Studies 1 and 2) takes an individual differences approach. Adopting a meta-analytic design with the addition of new data, we find that deliberate thinking and systematic thinking are differentially associated with personality traits (openness to experience with deliberate thinking; conscientiousness with systematic thinking) and with personal values (self-direction vs. conformity with deliberate thinking; security vs. stimulation with systematic thinking). Our second paradigm (Studies 3 and 4) employs a decision-making task (choosing between different problem types and levels of difficulty) to test for deliberate and systematic thinking in isolation from each other. We show that systematic thinking (in oneself and others) predicts a selection of rule-based over context-based problems, while deliberate thinking predicts a selection of difficult over simple problems. Our third paradigm (Studies 5 and 6) takes a cultural perspective. We show that although deliberate thinking is universally perceived as signifying competence, the contribution of systematic thinking to perceptions of competence is culturally dependent, differing for participants under a collectivistic versus individualistic mindset. Together our findings highlight the need to distinguish between deliberate and systematic thinking and underscore the need for studies of intuitive versus nonintuitive thinking to take a multidimensional perspective. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Genetic Association of a Gain-of-Function IFNGR1 Polymorphism and the Intergenic Region LNCAROD/DKK1 With Behçet's Disease.
Behçet's disease is a complex systemic inflammatory vasculitis of incompletely understood etiology. This study was undertaken to investigate genetic associations with Behçet's disease in a diverse multiethnic population. A total of 9,444 patients and controls from 7 different populations were included in this study. Genotyping was performed using an Infinium ImmunoArray-24 v.1.0 or v.2.0 BeadChip. Analysis of expression data from stimulated monocytes, and epigenetic and chromatin interaction analyses were performed. We identified 2 novel genetic susceptibility loci for Behçet's disease, including a risk locus in IFNGR1 (rs4896243) (odds ratio [OR] 1.25; P = 2.42 × 10-9 ) and within the intergenic region LNCAROD/DKK1 (rs1660760) (OR 0.78; P = 2.75 × 10-8 ). The risk variants in IFNGR1 significantly increased IFNGR1 messenger RNA expression in lipopolysaccharide-stimulated monocytes. In addition, our results replicated the association (P < 5 × 10-8 ) of 6 previously identified susceptibility loci in Behçet's disease: IL10, IL23R, IL12A-AS1, CCR3, ADO, and LACC1, reinforcing the notion that these loci are strong genetic factors in Behçet's disease shared across ancestries. We also identified >30 genetic susceptibility loci with a suggestive level of association (P < 5 × 10-5 ), which will require replication. Finally, functional annotation of genetic susceptibility loci in Behçet's disease revealed their possible regulatory roles and suggested potential causal genes and molecular mechanisms that could be further investigated. We performed the largest genetic association study in Behçet's disease to date. Our findings reveal novel putative functional variants associated with the disease and replicate and extend the genetic associations in other loci across multiple ancestries.


Adherence to process of care quality indicators after percutaneous coronary intervention in Ontario, Canada: a retrospective observational cohort study.
Public reporting of percutaneous coronary intervention (PCI) outcomes has been established in many jurisdictions to ensure optimal delivery of care. The majority of PCI report cards examine in-hospital mortality, but relatively little is known regarding the adherence to processes of care. A modified Delphi panel comprising cardiovascular experts was assembled to develop a set of PCI quality indicators. Indicators such as prescription of aspirin, dual antiplatelet therapy, statins and smoking cessation counselling were identified to represent high-quality PCI care. Chart abstraction was performed at 13 PCI hospitals in Ontario, Canada from 2009 to 2010 with at least 200 PCI patients randomly selected from each hospital. Our study sample included 3041 patients, of whom 18% had stable coronary artery disease (CAD) and 82% had an acute coronary syndrome (ACS). Their mean age was 63±12.4 years and 29% of patients were female. Prior to PCI, 89% were prescribed aspirin, and after PCI 98.7% were prescribed aspirin, 95.1% were prescribed dual antiplatelet therapy for 12 months after drug-eluting stents, and 94.9% were prescribed statins. The lowest performing quality indicator was smoking cessation counselling, observed in only 42% of current and past smokers (18% in patients with stable CAD and 47% in ACS). Our study demonstrates high levels of adherence to most quality indicators for patients undergoing PCI procedures in Ontario. In conclusion, smoking cessation counselling was not consistently performed across hospitals and represents an opportunity for future quality improvement efforts.


The MLL3/MLL4 branches of the COMPASS family function as major histone H3K4 monomethylases at enhancers.
Histone H3 lysine 4 (H3K4) can be mono-, di-, and trimethylated by members of the COMPASS (complex of proteins associated with Set1) family from Saccharomyces cerevisiae to humans, and these modifications can be found at distinct regions of the genome. Monomethylation of histone H3K4 (H3K4me1) is relatively more enriched at metazoan enhancer regions compared to trimethylated histone H3K4 (H3K4me3), which is enriched at transcription start sites in all eukaryotes. Our recent studies of Drosophila melanogaster demonstrated that the Trithorax-related (Trr) branch of the COMPASS family regulates enhancer activity and is responsible for the implementation of H3K4me1 at these regions. There are six COMPASS family members in mammals, two of which, MLL3 (GeneID 58508) and MLL4 (GeneID 8085), are most closely related to Drosophila Trr. Here, we use chromatin immunoprecipitation-sequencing (ChIP-seq) of this class of COMPASS family members in both human HCT116 cells and mouse embryonic stem cells and find that MLL4 is preferentially found at enhancer regions. MLL3 and MLL4 are frequently mutated in cancer, and indeed, the widely used HCT116 cancer cell line contains inactivating mutations in the MLL3 gene. Using HCT116 cells in which MLL4 has also been knocked out, we demonstrate that MLL3 and MLL4 are major regulators of H3K4me1 in these cells, with the greatest loss of monomethylation at enhancer regions. Moreover, we find a redundant role between Mll3 (GeneID 231051) and Mll4 (GeneID 381022) in enhancer H3K4 monomethylation in mouse embryonic fibroblast (MEF) cells. These findings suggest that mammalian MLL3 and MLL4 function in the regulation of enhancer activity and that mutations of MLL3 and MLL4 that are found in cancers could exert their properties through malfunction of these Trr/MLL3/MLL4-specific (Trrific) enhancers.


Hydrogen sulfide increases nitric oxide production from endothelial cells by an akt-dependent mechanism.
Hydrogen sulfide (H(2)S) and nitric oxide (NO) are both gasotransmitters that can elicit synergistic vasodilatory responses in the in the cardiovascular system, but the mechanisms behind this synergy are unclear. In the current study we investigated the molecular mechanisms through which H(2)S regulates endothelial NO production. Initial studies were performed to establish the temporal and dose-dependent effects of H(2)S on NO generation using EPR spin trapping techniques. H(2)S stimulated a twofold increase in NO production from endothelial nitric oxide synthase (eNOS), which was maximal 30 min after exposure to 25-150 μM H(2)S. Following 30 min H(2)S exposure, eNOS phosphorylation at Ser 1177 was significantly increased compared to control, consistent with eNOS activation. Pharmacological inhibition of Akt, the kinase responsible for Ser 1177 phosphorylation, attenuated the stimulatory effect of H(2)S on NO production. Taken together, these data demonstrate that H(2)S up-regulates NO production from eNOS through an Akt-dependent mechanism. These results implicate H(2)S in the regulation of NO production in endothelial cells, and suggest that deficiencies in H(2)S signaling can directly impact processes regulated by NO.


What is the difference between the inferior labial artery and the horizontal labiomental artery?
The inferior labial artery (ILA) and horizontal labiomental artery (HLA) can be regarded as the main arteries used in the lower lip pedicle for the perioral reconstruction. However, the courses of the ILA and HLA are described in diverse ways, and there is no obvious standard for distinguishing between them. The aim of this study was to elucidate the distribution patterns of the ILA and HLA, and the significance of the HLA in the vascularization of the lower lip. Sixty-three hemifaces from 18 Korean and 19 Thai cadavers were used in this study. The distribution patterns of the arteries of the lower lip area were classified based on the assumption that the HLA travels in the middle of the lower lip area, while the ILA runs along the lower lip border. The arterial distribution pattern of the HLA was classified into three types (I-III): type I, where the HLA ran horizontally in the lower lip area; type II, where the HLA curved upward to the vermilion border of the lower lip; and type III, where the HLA bifurcated into two branches. The ILA was also classified into three types (A-C): type A, where the ILA was not observed; type B, ILA was ramified from facial artery at the level of mouth corner; and type C, ILA arose from the superior labial artery (SLA) and supplied the lower lip vermillion border. Types I, II, and III were observed in 52.4 % (33/63), 39.7 % (25/63), and 7.9 % (5/63) of cases, respectively; and types A, B, and C were observed in 52.4 % (33/63), 36.5 % (23/63), and 11.1 % (7/63) of cases. Consideration of the two artery classifications together revealed seven types: type IA (14.3 %, 9/63), type IB (28.6 %, 18/63), type IC (9.5 %, 6/63), type IIA (30.2 %, 19/63), type IIB (7.9 %, 5/63), type IIC (1.6 %, 1/63), and type III (7.9 %, 5/63). The distance between the HLA and the midpoint between the mouth corner and the inferior mandibular margin was 0.4 ± 3.1 mm (mean ± SD) inferior to this point. The ILA and HLA should be differentiated according to the location of their origin rather than their terminating areas. The HLA could be considered as the main artery supplying the lower lip mucosa.


Mechanism of Water Content on the Electrochemical Surface Area of the Catalyst Layer in the Proton Exchange Membrane Fuel Cell.
Combined molecular dynamics (MD) simulation and experiment are adopted to gain the mechanism of water content on the electrochemical surface area (ECSA) of the catalyst layer in a proton exchange membrane fuel cell. The morphology of water domains in the catalyst layer has a strong impact on the ECSA via MD simulation. The morphology of the water domains is isolated water clusters at low water content, resulting in the poor ECSA due to the lack of proton transport paths. The transport paths of protons tend to be quickly established with increasing water content during the transition process of the morphology of water domains from isolated water clusters to the water channel network, thereby leading to the rapid increase of the ECSA. However, the slight increase of the ECSA at high water content mainly results from the improved contact area between water domains and Pt particle instead of the formation of new transport paths. In addition, the stronger binding of water molecules and the Pt particle at low temperature results in a higher ECSA.


Sildenafil for male erectile dysfunction: a systematic review and meta-analysis.
To determine the efficacy and safety of sildenafil citrate in the treatment of male erectile dysfunction. The MEDLINE, HealthSTAR, Current Contents, and Cochrane Library databases (January 1, 1995, through December 31, 2000); bibliographies of retrieved articles and review articles; conference proceedings abstracts; the Food and Drug Administration Web site; and the manufacturer. Trials were eligible if they included men with erectile dysfunction, compared sildenafil with control, were randomized, were of at least 7 days' duration, and assessed clinically relevant outcomes. Two reviewers independently evaluated study quality and extracted data in a standardized fashion. Twenty-seven trials (6659 men) met the inclusion criteria. In results pooled from 14 parallel-group, flexible as-needed dosing trials, sildenafil was more likely than placebo to lead to successful sexual intercourse, with a higher percentage of successful intercourse attempts (57% vs 21%; weighted mean difference, 33.7; 95% confidence interval [CI], 29.2-38.2; 2283 men) and a greater percentage of men experiencing at least 1 intercourse success during treatment (83% vs 45%; relative benefit increase, 1.8; 95% CI, 1.7-1.9; 2205 men). In data pooled from 6 parallel-group, fixed-dose trials, efficacy appeared slightly greater at higher doses. Treatment response appeared to vary between patient subgroups, although relative to placebo, sildenafil significantly improved erectile function in all evaluated subgroups. In trials with parallel-group design and flexible dosing, men randomized to receive sildenafil were less likely than those receiving placebo to drop out for any reason and no more likely to drop out due to an adverse event or laboratory abnormality. Specific adverse events with sildenafil included flushing (12%), headache (11%), dyspepsia (5%), and visual disturbances (3%); all adverse events were significantly less likely to occur with placebo. Sildenafil was not significantly associated with serious cardiovascular events or death. Sildenafil improves erectile function and is generally well tolerated. Treatment response seems to vary between patient subgroups, although sildenafil has greater efficacy than placebo in all evaluated subgroups.


Porcn-dependent Wnt signaling is not required prior to mouse gastrulation.
In mice and humans the X-chromosomal porcupine homolog (Porcn) gene is required for the acylation and secretion of all 19 Wnt ligands and thus represents a bottleneck for all Wnt signaling. We have generated a mouse line carrying a floxed allele for Porcn and used zygotic, oocyte-specific and visceral endoderm-specific deletions to investigate embryonic and extra-embryonic requirements for Wnt ligand secretion. We show that there is no requirement for Porcn-dependent secretion of Wnt ligands during preimplantation development of the mouse embryo. Porcn-dependent Wnts are first required for the initiation of gastrulation, where Porcn function is required in the epiblast but not the visceral endoderm. Heterozygous female embryos, which are mutant in both trophoblast and visceral endoderm due to imprinted X chromosome inactivation, complete gastrulation but display chorio-allantoic fusion defects similar to Wnt7b mutants. Our studies highlight the importance of Wnt3 and Wnt7b for embryonic and placental development but suggest that endogenous Porcn-dependent Wnt secretion does not play an essential role in either implantation or blastocyst lineage specification.


Resistance exercise and strong healthy children: safe when done right!
Weight lifting in children and youth is often viewed as a potentially harmful activity that could result in damage to bones, connective tissue, and muscle. Reviews, such as the one appearing in this issue of the journal by Behm et al. (Appl. Physiol. Metab. Nutr. 33: this issue), show that the balance of evidence indicates that weight lifting in pediatric populations is safe. Importantly, weight lifting, when performed in a safe and age-appropriate manner, is very beneficial on a number of health fronts, including strength and balance, self-esteem, and reducing cardiovascular risks. This is an understudied area that is still lacking in key areas of research to establish efficacy, dose-response relationships, and other health benefits.


Conformable Hybrid Systems for Implantable Bioelectronic Interfaces.
Conformable bioelectronic systems are promising tools that may aid the understanding of diseases, alleviate pathological symptoms such as chronic pain, heart arrhythmia, and dysfunctions, and assist in reversing conditions such as deafness, blindness, and paralysis. Combining reduced invasiveness with advanced electronic functions, hybrid bioelectronic systems have evolved tremendously in the last decade, pushed by progress in materials science, micro- and nanofabrication, system assembly and packaging, and biomedical engineering. Hybrid integration refers here to a technological approach to embed within mechanically compliant carrier substrates electronic components and circuits prepared with traditional electronic materials. This combination leverages mechanical and electronic performance of polymer substrates and device materials, respectively, and offers many opportunities for man-made systems to communicate with the body with unmet precision. However, trade-offs between materials selection, manufacturing processes, resolution, electrical function, mechanical integrity, biointegration, and reliability should be considered. Herein, prominent trends in manufacturing conformable hybrid systems are analyzed and key design, function, and validation principles are outlined together with the remaining challenges to produce reliable conformable, hybrid bioelectronic systems.


The role of theophyllines in a preventive approach for subjects with both mild and severe asthma.
Theophyllines continue to have a definite place in asthma therapy. With careful attention to therapeutic serum theophylline concentrations they are effective to improve spirometry and prevent symptoms. Theophyllines provide an additional benefit when prescribed with beta 2 agonists, and oral or inhaled steroids. Evening dosing with the once-daily, 24 hour formulation of theophylline 400 mg (Uniphyl) improves early morning function and prevents all or most of the early morning symptom exacerbation. Whereas there has been considerable focus on the potential side effects and theophylline toxicity, if subjects are introduced slowly to theophyllines, particularly in naive patients, and with careful attention to patient tolerance and serum theophylline concentrations, a majority of patients can be satisfactorily introduced to the products. With clinical experience, and when introduced with care, most patients will receive significant therapeutic benefit from theophyllines, with minimal or no side effects. Most patients readily accept the products, particularly the once daily 24 hour formulation.


A phylogenomic framework for assessing the global emergence and evolution of clonal complex 398 methicillin-resistant Staphylococcus aureus.
Distinct clones of methicillin-resistant Staphylococcus aureus (MRSA) have emerged as important causes of infection in individuals who have exposure to livestock (livestock-associated MRSA; LA-MRSA). Clonal complex 398 (CC398) is the most prevalent LA-MRSA clone, and has been reported from several geographical settings, including Europe, the Americas and Asia. To understand the factors contributing to the global dissemination of this clone, we analysed CC398 MRSA isolates from New Zealand (NZ), a geographically isolated country with an economy strongly dependent on livestock farming. We supplemented the NZ CC398 MRSA collection with global datasets of CC398 MRSA and CC398 methicillin-susceptible S. aureus. Here, we demonstrate multiple sporadic incursions of CC398 MRSA into NZ, as well as recent importation and spread of a swine-associated clade related to the European LA-MRSA lineage. Within a larger global phylogenomic framework, Bayesian modelling suggested that this NZ clade emerged in the late 2000s, with a probable origin in swine from Western Europe. Elucidating the factors responsible for the incursion and spread of LA-MRSA in geographically distant regions, such as NZ, provides important insights into global pathways of S. aureus transmission, and will inform strategies to control importation and spread.


Obesity-Related Hormones in Low-Income Preschool-Age Children: Implications for School Readiness.
Mechanisms underlying socioeconomic disparities in school readiness and health outcomes, particularly obesity, among preschool-aged children are complex and poorly understood. Obesity can induce changes in proteins in the circulation that contribute to the negative impact of obesity on health; such changes may relate to cognitive and emotion regulation skills important for school readiness. We investigated obesity-related hormones, body mass index (BMI), and school readiness in a pilot study of low-income preschoolers attending Head Start (participating in a larger parent study). We found that the adipokine leptin was related to preschoolers' BMI z-score, the appetite-regulating hormones ghrelin and glucagon-like peptide 1 (GLP-1), and pro-inflammatory cytokines typically associated with early life stress; and that some of these obesity-related biomarkers were in turn related to emotion regulation. Future work should evaluate how obesity may affect multiple domains of development, and consider modeling common physiological pathways related to stress, health, and school readiness.


[Effect of nonmutagenic doses of 1,4-bis-diazoacetylbutane on Streptomyces griseus].
Streptomyces griseus 15 was subjected to the action of 1,4-bis-diazoacetyl butane (DAB) taken at concentrations of 10 to 50,000 micrograms/ml. Small doses (10-100 micrograms/ml) of DAB had no mutagenic action and activated the cultural growth (the viability and the survival rate of spores increased on solid media, while the biomass yield rised in liquid media). Experiments were conducted using the method of orthogonal planning of a bifactorial experiment, and the role of the exposure time rised with a decrease in the mutagen concentration.


Physiological and pathological regulation of ACE2, the SARS-CoV-2 receptor.
The renin-angiotensin system (RAS) is crucial for the physiology and pathology of all the organs. Angiotensin-converting enzyme 2 (ACE2) maintains the homeostasis of RAS as a negative regulator. Recently, ACE2 was identified as the receptor of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the coronavirus that is causing the pandemic of Coronavirus disease 2019 (COVID-19). Since SARS-CoV-2 must bind with ACE2 before entering the host cells in humans, the distribution and expression of ACE2 may be critical for the target organ of the SARS-CoV-2 infection. Moreover, accumulating evidence has demonstrated the implication of ACE2 in the pathological progression in tissue injury and several chronic diseases, ACE2 may also be essential in the progression and clinical outcomes of COVID-19. Therefore, we summarized the expression and activity of ACE2 in various physiological and pathological conditions, and discussed its potential implication in the susceptibility of SARS-CoV-2 infection and the progression and prognosis of COVID-19 patients in the current review.


Protein splicing of the Saccharomyces cerevisiae VMA intein without the endonuclease motifs.
The protein splicing element (intein) of the vacuolar ATPase subunit (VMA) of Saccharomyces cerevisiae catalyzes both protein splicing and site-specific DNA cleavage. It has been demonstrated that the conserved splice junction residues are directly involved in protein splicing and the central dodecapeptide motifs are required for DNA cleavage. To examine whether the splicing activity of the intein can be structurally separated from the endonuclease motifs, we made large in-frame deletions at the central region of the intein. We demonstrate for the first time that protein splicing can proceed efficiently after the removal of the central region of the intein including the endonuclease motifs. Our results suggest that the N- and C-terminal regions of the Sce VMA intein may form a separate domain that is not only catalytically sufficient for protein splicing but also structurally independent from the endonuclease domain.


Acalculous and clostridial cholecystitis in a pig.
A 21-month-old domestic Hanford pig (Sus scrofa domestica) in a 1-year study for experimental myocardial infarction was euthanized at the end of the study. One week earlier, the animal had symptoms and elevated clinical chemistry results suggestive of hepatobiliary disease, which resolved after medical therapy. At necropsy, the gallbladder was markedly enlarged, discolored, and had a thickened wall. Within the gallbladder, there was abundant friable green-brown material. A culture of the gallbladder luminal material yielded Clostridium perfringens type A. Histopathology of the gallbladder demonstrated multifocal areas of necrosis of varying depths, admixed with an inflammatory infiltrate that was also observed on the serosa and within the associated adipose tissue. Luminal material was composed of cellular debris and bile sludge admixed with numerous bacterial rods. Smooth-muscle hypertrophy of numerous small arterioles with narrowed lumina was observed in the gallbladder. A diagnosis of acalculous cholecystitis presumably because of ischemia of the gallbladder with secondary clostridial infection was made. To the authors' knowledge, this is the first reported case of acalculous cholecystitis with evidence of vascular compromise in a pig, as well as cholecystitis secondarily attributed to Clostridium perfringens type A.


A statistical method for identifying differential gene-gene co-expression patterns.
To understand cancer etiology, it is important to explore molecular changes in cellular processes from normal state to cancerous state. Because genes interact with each other during cellular processes, carcinogenesis related genes may form differential co-expression patterns with other genes in different cell states. In this study, we develop a statistical method for identifying differential gene-gene co-expression patterns in different cell states. For efficient pattern recognition, we extend the traditional F-statistic and obtain an Expected Conditional F-statistic (ECF-statistic), which incorporates statistical information of location and correlation. We also propose a statistical method for data transformation. Our approach is applied to a microarray gene expression dataset for prostate cancer study. For a gene of interest, our method can select other genes that have differential gene-gene co-expression patterns with this gene in different cell states. The 10 most frequently selected genes, include hepsin, GSTP1 and AMACR, which have recently been proposed to be associated with prostate carcinogenesis. However, genes GSTP1 and AMACR cannot be identified by studying differential gene expression alone. By using tumor suppressor genes TP53, PTEN and RB1, we identify seven genes that also include hepsin, GSTP1 and AMACR. We show that genes associated with cancer may have differential gene-gene expression patterns with many other genes in different cell states. By discovering such patterns, we may be able to identify carcinogenesis related genes.


Phosphorylation potential and adenosine release during norepinephrine infusion in guinea pig heart.
This study tested the hypothesis that adenosine released from isolated guinea pig hearts (n = 5) in response to norepinephrine is related to the cellular phosphorylation potential (PP; [ATP]/[ADP][Pi]), where Pi is inorganic phosphate. 31P-nuclear magnetic resonance (NMR) was used to measure the relative concentrations of Pi, phosphocreatine (PCr), and ATP. Hearts were Langendorff perfused with a physiological salt solution containing 0.1 mM Pi. The venous effluent was collected for measurement of adenosine and partial pressure of oxygen (PO2). After a control period, norepinephrine (6 X 10(-8) M) was infused for 20 min during which 31P-NMR spectra and samples of venous effluent were collected every minute. With norepinephrine infusion, PCr decreased rapidly to 72% of control (P less than 0.05) by 8 min and then recovered to 80% of control for the remaining 12 min. ATP fell slowly to 70% of control (P less than 0.01) over 20 min. Pi increased to a peak at 2 min (P less than 0.01), then declined slowly to a steady state (60% of the peak and 3.5 X control) from 8 to 20 min. Adenosine release increased from 11 +/- 6 to a peak of 250 +/- 68 pmol.min-1.g-1 (P less than 0.01) at 7 min and then slowly fell (P less than 0.05) to a steady state of approximately 110 pmol.min-1.g-1 (P less than 0.01 vs. control) from 10 to 20 min.(ABSTRACT TRUNCATED AT 250 WORDS)


Immune regulation by CTLA-4--relevance to autoimmune diabetes in a transgenic mouse model.
The importance of cytotoxic T lymphocyte antigen-4 (CTLA-4) in immune regulation is unquestioned, yet a precise understanding of which cells express it, and how it mediates immune inhibitory function, is lacking. Regulatory T cells are known to constitutively express CTLA-4 intracellularly, whereas conventional T cells require activation to trigger CTLA-4 expression. However comparative analysis of CTLA-4 trafficking in regulatory and conventional subsets has not been performed. Here we assess CTLA-4 expression in antigen-specific conventional and regulatory cells responding to immunizing antigen in vivo and analyse the membrane trafficking of CTLA-4 using an in vitro recycling assay. We assess the expression of CTLA-4 on Treg infiltrating the pancreas in the DO11×RIP-mOVA diabetes model and the role of CTLA-4 in Treg function. Regulatory T cells show an enhanced capacity to traffic CTLA-4 following stimulation compared with conventional T cells. Treg infiltrating the pancreas in DO11×RIP-mOVA mice show high expression of CTLA-4. Furthermore CTLA-4-deficient Treg fail to control diabetes in an adoptive transfer model of diabetes, even in situations where they outnumber the disease-inducing conventional T cells. These data show that not only do regulatory T cells express higher levels of intracellular CTLA-4 than conventional T cells, but they also show an increased capacity to traffic CTLA-4 to the cell surface following stimulation. CTLA-4 is strongly upregulated in regulatory T cells infiltrating the target tissue in a mouse model of type 1 diabetes and expression of this protein is critical for effective regulation.


Cellular La protein shields nonsegmented negative-strand RNA viral leader RNA from RIG-I and enhances virus growth by diverse mechanisms.
The La antigen (SS-B) associates with a wide variety of cellular and viral RNAs to affect gene expression in multiple systems. We show that La is the major cellular protein found to be associated with the abundant 44-nucleotide viral leader RNA (leRNA) early after infection with respiratory syncytial virus (RSV), a nonsegmented negative-strand RNA virus. Consistent with this, La redistributes from the nucleus to the cytoplasm in RSV-infected cells. Upon RNA interference knockdown of La, leRNA is redirected to associate with the RNA-binding protein RIG-I, a known activator of interferon (IFN) gene expression, and this is accompanied by the early induction of IFN mRNA. These results suggest that La shields leRNA from RIG-I, abrogating the early viral activation of type I IFN. We mapped the leRNA binding function to RNA recognition motif 1 of La and showed that while wild-type La greatly enhanced RSV growth, a La mutant defective in RSV leRNA binding also did not support RSV growth. Comparative studies of RSV and Sendai virus and the use of IFN-negative Vero cells indicated that La supports the growth of nonsegmented negative-strand RNA viruses by both IFN suppression and a potentially novel IFN-independent mechanism.


Randomized comparative study between buccal mucosal and acellular bladder matrix grafts in complex anterior urethral strictures.
Urethral strictures have been a reconstructive dilemma for many years due to the limited availability of tissue substitutes and incidence of recurrence. Buccal mucosal grafts have been a favored material in instances where penile skin is unavailable due to its durability and excellent graft survival. Recently collagen based matrices derived from the bladder have been used successfully in patients with stricture disease and hypospadias. We performed a randomized comparative study to assess the outcome of the acellular bladder matrix compared to buccal mucosa in patients with complex urethral strictures. Human demineralized bone matrix, obtained from cadaveric donors, was processed and prepared for use as an off-the-shelf material. Thirty patients with stricture 21 to 59 years old (mean 36.2) were enrolled and assessed using a standard protocol. The stricture length ranged from 2 to 18 cm (mean 6.9), of which 11 patients had bulbar, 7 had pendulous and 12 had combined bulbopendulous strictures. Of the 30 patients 7 had received no previous intervention while the remaining 23 had undergone 1 to 7 procedures (mean 1.9). All patients were randomized and alternatively assigned to receive either buccal mucosa or decellularized bladder [corrected] matrix and underwent an onlay procedure. All patients except 2 who were lost during followup were followed for 18 to 36 months (mean 25). In patients with a healthy urethral bed (less than 2 prior operations) the success rate of buccal mucosa grafts (10 of 10) was similar to the bladder matrix grafts (8 of 9) in terms of patency. In patients with an unhealthy urethral bed (more than 2 prior operations) only 2 of 6 patients with a bladder matrix graft were successful, whereas all 5 patients with a buccal mucosa graft had a patent urethra. Postoperative uroflowmetry showed significant voiding improvement in both groups. Histology of the graft biopsies showed normal urethral tissue characteristics. This study demonstrates that the use of acellular bladder matrix is a viable option for urethral repair. Demineralized bone matrix as an off-the-shelf biomaterial achieves the best results in patients with a healthy urethral bed, no spongiofibrosis and good urethral mucosa.


Adenosine amplifies follicle-stimulating hormone action in granulosa cells and luteinizing hormone action in luteal cells of rat and human ovaries.
It is known that adenosine amplifies LH-stimulated cAMP accumulation and progesterone production in rat luteal cells. The aim of this study was to investigate the effect of adenosine on cAMP and steroid production in short term cultures of rat and human luteal cells in the presence of LH and in rat and human granulosa cells in the presence of FSH. In rat luteal cells, the maximal cAMP response to LH and adenosine amplification of this response occurred in the midluteal phase and was decreased in both early and late luteal cells. In humans, adenosine (50 microM) increased cAMP and and progesterone accumulation by 100% and 40%, respectively, in periovulatory granulosa cells. Adenosine also amplified cAMP accumulation in response to increasing hCG concentrations by 2- to 3-fold in human luteal cells. The ability of the human luteal cell to respond to hCG with cAMP accumulation and the ability of adenosine to amplify this cAMP response appeared to be inversely related to human luteal cell age. In isolated preparations of rat granulosa cells, adenosine amplified cAMP accumulation in response to FSH, and cAMP accumulation was inversely proportional to the duration of follicular development. In human periovulatory granulosa cells, adenosine (100 microM0 increased cAMP accumulation by 2-fold and amplified FSH-stimulated cAMP accumulation by 25%. These studies suggest that in both the rat and human, adenosine may physiologically affect gonadotropin function in both follicular and luteal ovarian tissue.


Salinization of the soil solution decreases the further accumulation of salt in the root zone of the halophyte Atriplex nummularia Lindl. growing above shallow saline groundwater.
Water use by plants in landscapes with shallow saline groundwater may lead to the accumulation of salt in the root zone. We examined the accumulation of Na+ and Cl- around the roots of the halophyte Atriplex nummularia Lindl. and the impacts of this increasing salinity for stomatal conductance, water use and growth. Plants were grown in columns filled with a sand-clay mixture and connected at the bottom to reservoirs containing 20, 200 or 400 mM NaCl. At 21 d, Na+ and Cl- concentrations in the soil solution were affected by the salinity of the groundwater, height above the water table and the root fresh mass density at various soil depths (P < 0.001). However, by day 35, the groundwater salinity and height above the water table remained significant factors, but the root fresh mass density was no longer significant. Regression of data from the 200 and 400 mM NaCl treatments showed that the rate of Na+ accumulation in the soil increased until the Na+ concentration reached ~250 mM within the root zone; subsequent decreases in accumulation were associated with decreases in stomatal conductance. Salinization of the soil solution therefore had a feedback effect on further salinization within the root zone.


Neuroprotective strategies with circulatory arrest in open aortic surgery - A meta-analysis.
Deep hypothermic circulatory arrest (DHCA) in aortic surgery is associated with morbidity and mortality despite evolving strategies. With the advent of antegrade cerebral perfusion (ACP), moderate hypothermic circulatory arrest (MHCA) was reported to have better outcomes than DHCA. There is no standardised guideline or consensus regarding the hypothermic strategies to be employed in open aortic surgery. Meta-analysis was performed comparing DHCA with MHCA + ACP in patients having aortic surgery. A systematic review of the literature was undertaken. Any studies with DHCA versus MHCA + ACP in aortic surgeries were selected according to specific inclusion criteria and analysed to generate summative data. Statistical analysis was performed using STATS Direct. The primary outcomes were hospital mortality and post-operative stroke. Secondary outcomes were cardiopulmonary bypass time (CPB), post-operative blood transfusion, length of ICU stay, respiratory complications, renal failure and length of hospital stay. Subgroup analysis of primary outcomes for Arch surgery alone was also performed. Fifteen studies were included with a total of 5869 patients. There was significantly reduced mortality (Pooled OR = +0.64, 95% CI = +0.49 to +0.83; p = 0.0006) and stroke rate (Pooled OR = +0.62, 95% CI = +0.49 to +0.79; p < 0.001) in the MHCA group. MHCA was associated significantly with shorter CPB times, shorter duration in ICU, less pulmonary complications, and reduced rates of sepsis. There was no statistical difference between the two groups in terms of circulatory arrest times, X-Clamp times, total operation duration, transfusion requirements, renal failure and post-op hospital stay. MHCA + ACP are associated with significantly better post-operative outcomes compared with DHCA for both mortality and stroke and majority of the secondary outcomes.


[Development of mono- and associative cultures of sulphate-reducing bacteria and formation of exopolymeric complex].
Differences were found in development of the monoculture of sulphate-reducing bacteria Desulfovibrio sp. Kyiv-10 and artificial associations of cultures of sulphate reducing bacteria with heterotrophic satellites under development in a form of biofilm or plankton. Correlation between exopolymers synthesis in the biofilm and steel corrosion velocity has been found. Glucose, galactose, mannose, xylose, rhibose and nonidentified carbohydrates were found in the composition of exopolymers synthesized by monoculture Desulfovibrio sp. Kyiv-10 under its development in a form of biofilm. The biofilm and plankton cells of associative cultures synthesized exopolysaccharide which included rhamnose, arabinose and fucose. Glucose content in biofilm EPM was considerably higher (41.0055.9%) than in EPM of plankton cells (24.7-32.8).


3D volumetric muscle modeling for real-time deformation analysis with FEM.
Computer simulation is promising numerical tool to study muscle volumetric deformations. However, most models are facing very long computation time and thus are based on simplified wire Hill muscle model. The purpose of this study is to develop a real-time three-dimensional biomechanical model of volumetric muscle based on modified Hill model for the active stress which is controlled from EMG recordings. Finite element model is used to estimate the passive behavior of the muscle and tendons during contraction. We demonstrate that this 3D model implementation is very cost effective with respect to the computation time and the simulation gives good results compared to real measured data. Thus, this effective implementation will allow implementing much more complex and realistic models considering the muscle as volumetric continuum, with moderate computation time.


Cargo-shell and cargo-cargo couplings govern the mechanics of artificially loaded virus-derived cages.
Nucleic acids are the natural cargo of viruses and key determinants that affect viral shell stability. In some cases the genome structurally reinforces the shell, whereas in others genome packaging causes internal pressure that can induce destabilization. Although it is possible to pack heterologous cargoes inside virus-derived shells, little is known about the physical determinants of these artificial nanocontainers' stability. Atomic force and three-dimensional cryo-electron microscopy provided mechanical and structural information about the physical mechanisms of viral cage stabilization beyond the mere presence/absence of cargos. We analyzed the effects of cargo-shell and cargo-cargo interactions on shell stability after encapsulating two types of proteinaceous payloads. While bound cargo to the inner capsid surface mechanically reinforced the capsid in a structural manner, unbound cargo diffusing freely within the shell cavity pressurized the cages up to ∼30 atm due to steric effects. Strong cargo-cargo coupling reduces the resilience of these nanocompartments in ∼20% when bound to the shell. Understanding the stability of artificially loaded nanocages will help to design more robust and durable molecular nanocontainers.


Is There a Role for Surgery in Patients with Neuroendocrine Tumors of the Esophagus? A Contemporary View from the NCDB.
Esophageal neuroendocrine tumors (eNETs) are exceedingly rare, aggressive and have a poor prognosis. Treatment guidelines are ill-defined and mainly based on evidence from case reports and analogous experiences drawn from similar disease sites. The NCDB was reviewed for histologically confirmed stage I-III, primary eNETs from 2006 to 2014. Patients were grouped into whether or not they underwent primary tumor resection. Univariate, multivariable, and full bipartite propensity score (PS) adjusted Cox regression analyses were used to assess overall and relative survival differences. A total of 250 patients were identified. Mean age was 65.0 (standard deviation [SD] 11.9) years, and 174 (69.6%) patients were male. Most patients had stage III disease (n = 136, 54.4%), and the most common type of NET was small cell eNET (n = 111, 44.4%). Chemotherapy was used in 186 (74.4%), radiation therapy in 178 (71.2%), and oncological resection was performed in 69 (27.6%) patients. Crude 2-year survival rates were higher in the operated (57.3%) compared with the nonoperated group (35.2%; p < 0.001). The survival benefit held true after multivariable adjustment (hazard ratio [HR] 0.47, 95% confidence interval [CI] 0.32-0.69, p < 0.001). After full bipartite PS adjustment analysis, survival was longer for patients who received a surgical resection compared with those who did not (HR 0.48, 95% CI 0.31-0.75, p = 0.003) with a corresponding 2-year overall survival rate of 63.3% (95% CI 52.0-77.2) versus 38.8% (95% CI 30.9-48.8), respectively. Multimodal treatment that includes surgery is associated with better overall survival for eNETs. Additional research is needed to more definitively identify patients who benefit from esophagectomy and to establish an appropriate treatment algorithm.


HIV-induced T-cell syncytia release a two component T-helper cell chemoattractant composed of Nef and Tat.
Using a newly developed gradient chamber to provide independent measurements of chemokinesis (stimulated motility) and chemotaxis (stimulated motility up a concentration gradient) of individual T-helper cells, it was recently demonstrated that HIV-induced T-cell syncytia release two distinct chemotactic activities that are separable by their rates of diffusion. The molecular masses of the two chemoattractant activities were estimated to be 30 and 120 kDa. The higher molecular mass activity was demonstrated to be the viral glycoprotein gp120. In an attempt to identify the lower molecular mass activity, chemotaxis and chemokinesis of T-helper cells were analyzed in individual concentration gradients of the virally encoded proteins Rev, p24, Tat and Nef. None functioned alone as a chemoattractant, but both Tat and Nef alone functioned as chemokinetic stimulants. When Tat and Nef were used together to generate parallel gradients, they stimulated chemotaxis. Antibody to either Tat or Nef neutralized the lower molecular mass chemotactic activity released by syncytia. The addition of antibody to the CD4 receptor or the addition of soluble CD4 inhibited high molecular mass chemotactic activity but not the low molecular mass chemotactic activity in HIV-induced syncytium-conditioned medium, demonstrating that the former but not the latter activity is mediated through the CD4 receptor. These results identify the combination of Nef and Tat as the lower molecular mass T cell chemoattractant released by HIV-induced syncytia, and provide the first evidence suggesting that parallel concentration gradients of two proteins are necessary for chemotaxis.


Factor Structure and Psychometric Properties of Emotional Eater Questionnaire (EEQ) in Spanish Colleges.
Emotional eating (EE) patterns have been shown to play a relevant role in the development of overweight problems. However, there is a gap in research aimed at validating questionnaires to assess EE in specific populations. The aim of the study was to analyze factor structure and psychometric properties of Emotional Eater Questionnaire (EEQ) in Spanish universities. EEQ, state-anxiety subscale of STAI and a questionnaire about health habits were filled out by 295 students. Exploratory Factor Analysis (EFA) by using Unweight Least Squares (ULS) method was carried out. To determine factor numbers we used eigenvalues, parallel analysis, and goodness of fit statistics. Cronbach's alpha and Spearman correlations were used to analyze reliability, convergent, and concurrent validity. The parallel analysis and goodness of fit statistics showed that unifactorial structure of seven items was the most appropriate what accounted for 57% of the variance. Internal consistency was good (α = 0.753), as well as convergent validity (r = 0.317; p < 0.001). Concurrent validity was significant for three of the five criteria (r = -0.224; p < 0.001 and r = -0.259; p < 0.001). The results suggest some differences in the structure of the psychometric assessment of EE in sub-clinical population in comparison with previous studies carried on with an overweight population, what could be relevant to obesity prevention.


Amyloid fibril formation and seeding by wild-type human lysozyme and its disease-related mutational variants.
Wild-type human lysozyme and its two stable amyloidogenic variants have been found to form partially folded states at low pH. These states are characterized by extensive disruption of tertiary interactions and partial loss of secondary structure. Incubation of the proteins at pH 2.0 and 37 degrees C (Ile56Thr and Asp67His variants) or 57 degrees C (wild-type) results in the formation of large numbers of fibrils over several days of incubation. Smaller numbers of fibrils could be observed under other conditions, including neutral pH. These fibrils were analyzed by electron microscopy, Congo red birefringence, thioflavine-T binding, and X-ray fiber diffraction, which unequivocally show their amyloid character. These data demonstrate that amyloidogenicity is an intrinsic property of human lysozyme and does not require the presence of specific mutations in its primary structure. The amyloid fibril formation is greatly facilitated, however, by the introduction of "seeds" of preformed fibrils to the solutions of the variant proteins, suggesting that seeding effects could be important in the development of systemic amyloidosis. Fibril formation by wild-type human lysozyme is greatly accelerated by fibrils of the variant proteins and vice versa, showing that seeding is not specific to a given protein. The fact that wild-type lysozyme has not been found in ex vivo deposits from patients suffering from this disease is likely to be related to the much lower population of incompletely folded states for the wild-type protein compared to its amyloidogenic variants under physiological conditions. These results support the concept that the ability to form amyloid is a generic property of proteins, but one that is mitigated against in a normally functioning organism.


Stroke Mimics in a Stroke Care Pathway Based on MRI Screening.
Since the use of tissue plasminogen activator for acute ischemic stroke (IS), stroke care pathways have been developed for patients with suspicion of acute stroke. The aim of this prospective observational study was to analyze the stroke mimic (SM) characteristics in patients who were part of our stroke care pathway. All consecutive patients admitted in the code stroke within a 1-year period were prospectively enrolled in this study. Patients with a sudden onset of neurological focal deficit in a time window less than 4H30 as indicated for intravenous thrombolysis, had been accepted in the pathway by a neurologist who was directly contactable by the prehospital emergency medical service 24 h per day. Patients arrived directly on the MRI site without passing by the emergency department. A clinical neurological evaluation and a brain MRI with tri-dimensional time-of-flight magnetic resonance angiography were performed. The FAST score was calculated a posteriori. The final discharge diagnosis was concluded either immediately after both neurological examination and cerebrovascular neuroimaging or after other relevant investigations. We classified the discharge diagnosis into neurovascular diseases (NVDs) and into SM. There were 1,361 consecutive patients admitted for suspicion of acute stroke. Sixty-two percent (n = 840) had an NVD including IS (n = 529), transient ischemic attacks (n = 236), intracranial hemorrhages (n = 68), cerebral venous thrombosis (n = 3) and neurovascular medullar pathologies (n = 4). SM represented 38% of cases (n = 521) and the most frequent discharge diagnosis was defined as headaches (18.6%), psychological disorders (16.7%), peripheral vertigo (11.9%) and epilepsy (10.6%). The comparison between the characteristics of the NVD and those of the SM groups showed some significant differences: in the SM group, women were more represented, patients were younger and the NIHSS was lower than in the NVD group. All cardiovascular risk factors were more represented in the NVD group. Concerning the symptoms, motor deficit, speech disturbances, homonymous lateral hemianopia and head and gaze deviation were more represented in the NVD group, whereas vertigo, non-systematized visual trouble, headache, confusion, weakness, neuropsychological symptoms, seizure and chest pain were significantly more frequent in the SM group. The negative predictive value of the FAST score was 64% and the positive predictive value was 76%. A rate of SM up to 38% of the code stroke system confirms the difficulty to distinguish clinically a stroke from another diagnosis. In this study, using cerebral MRI in first intention was of special interest in patients with acute neurological symptoms to differentiate an NVD from an SM.


Insights into maleimide-thiol conjugation chemistry: Conditions for efficient surface functionalization of nanoparticles for receptor targeting.
Maleimide-thiol chemistry is widely used for the design and preparation of ligand-decorated drug delivery systems such as poly(lactide-co-glycolide) (PLGA) based nanoparticles (NPs). While many publications on nanocarriers functionalized exploiting this strategy are available in the literature, the conditions at which this reaction takes place vary among publications. This paper presents a comprehensive study on the conjugation of the peptide cRGDfK and the nanobody 11A4 (both containing a free thiol group) to maleimide functionalized PLGA NPs by means of the maleimide-thiol click reaction. The influence of different parameters, such as the nanoparticles preparation method and storage conditions as well as the molar ratio of maleimide to ligand used for conjugation, on the reaction efficiency has been evaluated. The NPs were prepared by a single or double emulsion method using different types and concentrations of surfactants and stored at 4 or 20 °C before reaction with the targeting moieties. Several maleimide to ligand molar ratios and different reaction times were studied and the conjugation efficiency was determined by quantification of the not-bound ligand by liquid chromatography. The kind of emulsion used to prepare the NPs as well as the type and concentration of surfactant used had no effect on the conjugation efficiency. Reaction between the maleimide groups present in the NPs and cRGDfK was optimal at a maleimide to thiol molar ratio of 2:1, reaching a conjugation efficiency of 84 ± 4% after 30 min at room temperature in 10 mM HEPES pH 7.0. For 11A4 nanobody the optimal reaction efficiency, 58 ± 12%, was achieved after 2 h of incubation at room temperature in PBS pH 7.4 using a 5:1 maleimide to protein molar ratio. Storage of the NPs at 4 °C for 7 days prior to their exposure to the ligands resulted in approximately 10% decrease in the reactivity of maleimide in contrast to storage at 20 °C which led to almost 40% of the maleimide being unreactive after the same storage time. Our findings demonstrate that optimization of this reaction, particularly in terms of reactant ratios, can represent a significant increase in the conjugation efficiency and prevent considerable waste of resources.


Transcranial direct current stimulation (tDCS) enhances internal source monitoring abilities in healthy participants.
Source monitoring refers to the ability to identify the origin of a memory, for example, whether you remember saying something or thinking about it, and confusions of these sources have been associated with the experience of auditory verbal hallucinations (AVHs). Both AVHs and source confusions are reported to originate from dysfunctional brain activations in the prefrontal cortex (PFC) and the superior temporal gyrus (STG); specifically, it is assumed that a hypoactive PFC and a hyperactive STG gives rise to AVHs and source confusions. We set out to test this assumption by trying to mimic this hypertemporal/hypofrontal model in healthy individuals with transcranial direct current stimulation (tDCS): the inhibitory cathode was placed over the left PFC and the excitatory anode over the left dorsolateral STG. Participants completed a reality monitoring task (distinguishing between external and internal memory sources) and an internal source monitoring task (distinguishing between two or more internal memory sources) in two separate experiments (offline vs. online tDCS). In the offline experiment (n = 34), both source monitoring tasks were completed after tDCS stimulation, and in the online experiment (n = 27) source monitoring tasks were completed while simultaneously being stimulated with tDCS. We found that internal source monitoring abilities were significantly enhanced during active online tDCS, while reality monitoring abilities were unaffected by stimulation in both experiments. We speculate, based on combining the present findings with previous studies, that there might be different brain areas involved in reality and internal source monitoring. While internal source monitoring seems to involve speech production areas, specifically Broca's area, as suggested in the present study, reality monitoring seems to rely more on the STG and DLPFC, as shown in other studies of the field.


Simultaneous flow injection preconcentration of lead and cadmium using cloud point extraction and determination by atomic absorption spectrometry.
A flow injection (FI) micelle-mediated separation/preconcentration procedure for the determination of lead and cadmium by flame atomic absorption spectrometry (FAAS) has been proposed. The analytes reacted with 1-(2-thiazolylazo)-2-naphthol (TAN) to form hydrophobic chelates, which were extracted into the micelles of 0.05% (w/v) Triton X-114 in a solution buffered at pH 8.4. In the preconcentration stage, the micellar solution was continuously injected into a flow system with four mini-columns packed with cotton, glass wool, or TNT compresses for phase separation. The analytes-containing micelles were eluted from the mini-columns by a stream of 3molL(-1) HCl solution and the analytes were determined by FAAS. Chemical and flow variables affecting the preconcentration of the analytes were studied. For 15mL of preconcentrated solution, the enhancement factors varied between 15.1 and 20.3, the limits of detection were approximately 4.5 and 0.75microgL(-1) for lead and cadmium, respectively. For a solution containing 100 and 10microgL(-1) of lead and cadmium, respectively, the R.S.D. values varied from 1.6 to 3.2% (n=7). The accuracy of the preconcentration system was evaluated by recovery measurements on spiked water samples. The method was susceptible to matrix effects, but these interferences were minimized by adding barium ions as masking agent in the sample solutions, and recoveries from spiked sample varied in the range of 95.1-107.3%.


A feasibility randomised controlled trial of a motivational interviewing-based intervention for weight loss maintenance in adults.
Obesity has significant health and NHS cost implications. Relatively small reductions in weight have clinically important benefits, but long-term weight loss maintenance (WLM) is challenging. Behaviour change interventions have been identified as key for WLM. Motivation is crucial to supporting behaviour change, and motivational interviewing (MI) has been identified as a successful approach to changing health behaviours. The study was designed as an adequately powered, pragmatic randomised controlled trial (RCT); however, owing to recruitment issues, the study became a feasibility trial. To assess recruitment, retention, feasibility, acceptability, compliance and delivery of a 12-month intervention to support WLM. Secondary objectives were to assess the impact of the intervention on body mass index (BMI) and other secondary outcomes. Three-arm individually randomised controlled trial comprising an intensive arm, a less intensive arm and a control arm. Community setting in South Wales and the East Midlands. Individuals aged 18-70 years with a current or previous BMI of ≥ 30 kg/m(2) who could provide evidence of at least 5% weight loss during the previous 12 months. Participants received individually tailored MI, which included planning and self-monitoring. The intensive arm received six face-to-face sessions followed by nine telephone sessions. The less intensive arm received two face-to-face sessions followed by two telephone sessions. The control arm received a leaflet advising them on healthy lifestyle. Feasibility outcomes included numbers recruited, retention and adherence. The primary effectiveness outcome was BMI at 12 months post randomisation. Secondary outcomes included waist circumference, waist-to-hip ratio, physical activity, proportion maintaining weight loss, diet, quality of life, health service resource usage, binge eating and well-being. A process evaluation assessed intervention delivery, adherence, and participants' and practitioners' views. Economic analysis aimed to assess cost-effectiveness in terms of quality-adjusted life-years (QALYs). A total of 170 participants were randomised. Retention was good (84%) and adherence was excellent (intensive, 83%; less intensive, 91%). The between-group difference in mean BMI indicated the intensive arm had BMIs 1.0 kg/m(2) lower than the controls [95% confidence interval (CI) -2.2 kg/m(2) to 0.2 kg/m(2)]. Similarly, a potential difference was found in weight (average difference of 2.8 kg, 95% CI -6.1 kg to 0.5 kg). The intensive arm had odds of maintaining on average 43% [odds ratio(OR) 1.4, 95% CI 0.6 to 3.5] higher than controls. None of these findings were statistically significant. Further analyses controlling for level of adherence indicated that average BMI was 1.2 kg/m(2) lower in the intensive arm than the control arm (95% CI -2.5 kg/m(2) to 0.0 kg/m(2)). The intensive intervention led to a statistically significant difference in weight (mean -3.7 kg, 95% CI -7.1 kg to -0.3 kg). The other secondary outcomes showed limited evidence of differences between groups. The intervention was delivered as planned, and both practitioners and participants were positive about the intervention and its impact. Although not powered to assess cost-effectiveness, results of this feasibility study suggest that neither intervention as currently delivered is likely to be cost-effective in routine practice. This is the first trial of an intervention for WLM in the UK, the intervention is feasible and acceptable, and retention and adherence were high. The main effectiveness outcome showed a promising mean difference in the intensive arm. Owing to the small sample size, we are limited in the conclusions we can draw. However, findings suggest that the intensive intervention may facilitate long-term weight maintenance and, therefore, further testing in an effectiveness trial may be indicated. Research examining WLM is in its infancy, further research is needed to develop our understanding of WLM and to expand theory to inform the development of interventions to be tested in rigorously designed RCTs with cost-effectiveness assessed. Current Controlled Trials ISRCTN35774128. This project was funded by the National Institute for Health Research Health Technology Assessment programme and will be published in full in Health Technology Assessment; Vol. 19, No. 50. See the NIHR Journals Library website for further project information.


Rotenone ameliorates chronic renal injury caused by acute ischemia/reperfusion.
Acute kidney injury (AKI) has been widely recognized as an important risk factor leading to the occurrence and progression of chronic kidney disease (CKD). Thus, development of the strategies in retarding the transition of AKI to CKD is becoming a hot research field. Recently, accumulating evidence suggested a pathogenic role of mitochondrial dysfunction in both AKI and CKD. Therefore, in the present study, we evaluated the effect of mitochondrial complex 1 inhibition by rotenone on the chronic renal damage induced by acute ischemia-reperfusion. The mice were treated with 45 min unilateral renal ischemia and reperfusion (I/R) to induce an acute renal injury. After three days of I/R injury, rotenone at a dose of 200 ppm in food was administered to the mice. Strikingly, after three weeks treatment with rotenone, we found that the unilateral I/R-induced tubular damage, tubulointerstitial fibrosis were all attenuated by rotenone as determined by the tubular injury score, Masson staining, and the levels of collagen-I, collagen-III, fibronectin, PAI-1, and TGF-β. Meanwhile, the enhanced inflammatory markers of TNF-α, IL-1β, IL-6, and IL-18 and apoptotic markers of Bax and caspase-3 were all significantly blunted by inhibiting mitochondrial complex-1. Moreover, rotenone treatment also partially protected the mitochondria as shown by the restoration of mitochondrial SOD (SOD2), ATPB, and mitochondrial DNA copy number. These findings suggested that inhibition of mitochondrial complex-1 activity by rotenone could retard the progression of AKI to CKD probably via protecting the mitochondrial function to some extent.


Internal validity of the revised HiMAT for people with neurological conditions.
The High-level Mobility Assessment Tool (HiMAT) was developed to measure high-level mobility limitations following traumatic brain injury. The aim of this study was to investigate if the revised HiMAT is valid for use with adults with neurological conditions other than traumatic brain injury. Cross-sectional study. Ninety-five participants with neurological conditions. HiMAT score sheets were retrieved from the central medical files of people who had attended a major rehabilitation facility for a neurological condition from January 2006 to October 2007. Additional HiMAT score sheets were submitted by therapists who participated in the HiMAT User's Group. Rasch analysis (RUMM2030 software) was used to determine the overall fit of the model, individual item fit and differential item functioning. Rasch analysis supported the internal validity of the revised eight-item HiMAT for individuals with neurological conditions. It showed good overall fit (P = 0.74), no misfitting items and excellent internal consistency (Person Separation Index = 0.91). The HiMAT is unidimensional with no evidence of response dependency and no differential item functioning for age or sex. Further development of the revised HiMAT is required to investigate other aspects of validity, reliability and responsiveness in different neurological populations. However, the results support the internal validity of the revised HiMAT when used for people with neurological conditions who are able to walk without gait aids.


Fatalities of children 0-7 years old in the second row.
The safety of children riding in the second-row of light vehicles remains an important issue. This study investigates fatal accidents of children 0-7 years old by seating position and principal direction of force. It considers the number of fatalities, exposure, and risk. The 1996-2005 FARS was analyzed for occupant fatalities by age (0-7 and >or= 8 years old), seating position (front, second, and third-row, and left, middle and right) and principal direction of force (1-12 o'clock PDOF, rollover, and other/unknown). Light vehicles were included with model year 1990+. The 1996-2005 NASS-CDS was similarly analyzed for occupant exposure. Fatality risk was defined as the number of fatalities in FARS divided by the exposure from NASS-CDS for each seating position and crash direction. Two thirds (67.8%) of 0-to 7-year-old child fatalities occupied second-row-seats in FARS. A nearly equal number died in rollovers (20.3%), front impacts (20.2%), and side crashes (19.6%). About 354 deaths occur to second-row-seated children annually. A majority (76.6%) of children ride in the second-row based on tow away crashes in NASS-CDS. Most are exposed to front impacts (41.6%), followed by side impacts (16.5%). Rollovers are the lowest frequency (5.0%). However, fatality risk is highest in rollovers (1.37%), followed by right-side (0.47%) and left-side impacts (0.34%). Near-seated children in the second-row have a 3.04% fatality risk for right-side (3 o'clock) impacts. This is nearly twice the 1.53% risk for near-side second-row children in left-side (9 o'clock) impacts. Overall, right-side impacts have a 37% higher fatality risk than left-side impacts, irrespective of second-row seating position. The second-row has a 43.4% lower fatality risk than the front seat (0.30% v 0.53%) and the third-row is 58.5% lower (0.22% v 0.53%) for 0-to 7-year-old children. Overall, children seated behind the driver have an 8.1% lower fatality risk than those seated behind the right-front passenger, although the center second-row-seat has the lowest risk (0.27%). Children in the second-row have a 65-71% lower fatality risk than the driver with the lowest relative risk in the center second-row-seat (0.29) and highest in the second-row right position (0.35). Children seated behind the driver have a lower fatality risk than those seated behind the right-front passenger. The higher risk to right second-row children is from near-side impacts at 3-4 o'clock and rollovers. In terms of priorities, rollovers, side impacts, and frontal crashes cause most fatalities. The highest risks for 0-to 7-year-old children in the second-row are in near-side impacts and rollovers.


Selective immunoglobulin A deficiency associated with primary biliary cirrhosis in a family with liver disease.
A family is described in which multiple members are afflicted with liver disease and primary biliary cirrhosis (PBC). In the third generation, one member died of PBC, and a second individual has both symptomatic PBC and selective immunoglobulin A (IgA) deficiency, an association not previously reported. By culturing this patient's lymphocytes in vitro it was shown that the IgA deficiency was due to a failure of B cells to secrete IgA. Two other siblings of this patient have multiple serum biochemical and serologic abnormalities that are sometimes associated with PBC, but they do not have histopathologically overt PBC or IgA deficiency. All three surviving family members have a diminished autologous mixed lymphocyte reaction, an immunologic abnormality that has previously been found in patients with PBC, selective IgA deficiency, and several autoimmune diseases. As there is an association between selective IgA deficiency and certain autoimmune diseases, it is possible that this immunodeficiency contributed to the development of PBC in the patient in whom the two diseases coexisted. Furthermore, the occurrence of PBC in a patient with selective IgA deficiency indicates that the pathogenesis of PBC does not require IgA-dependent immune mechanisms.


Visual pigment fluorescence.
The fluorescence of cattle rod outer segments (dried) and of rhodopsin in solution lies in the range of 575 to 600 millimicrons with a quantum efficiency of 0.005 if excitation is in the visible band near 500 millimicrons. The emission is abolished by bleaching at -196 degrees C but can be reversibly regenerated by irradiation with light of longer wavelength (600 millimicrons). This behavior reflects the known interconversion of rhodopsin to prelumirhodopsin at this temperature.


Predictors and mechanisms of oncological failure following nephron-sparing surgery for renal cancer.
One of the major concerns in performing nephron-sparing surgery (NSS) for renal cell carcinoma (RCC) is the risk of tumor recurrence. To assess the rate, predictors and mechanisms of oncological failure in patients after NSS for renal cancer. Between 1993 and 2008 NSS was performed in 229 patients via flank incision. Only patients without metastases at diagnosis and minimal 12 months follow-up were included in the outcome analysis. During a mean follow-up of 45 +/- 34 months (range 6-168 months) tumor recurrence was observed in 13 patients (5.6%). Mean follow-up time for detection of oncological failure was 51 months (range 6-132 months). All patients with oncological failure were males, with a mean age of 61 years (median 58, range 51-74 years). The average size of the enucleated lesion was 5 cm (range 4-7 cm). Intraoperative frozen sections as well as postoperative final pathological examination of the surgical margins were negative in all recurrent cases. Mechanisms of recurrence were distant metastases (n=4), surgical scar implantation (n=2), perirenal fat recurrence (n=2), local renal recurrence at the surgical site (n=1), and new renal lesions (n=4). Predictors of oncological failure included warm ischemia time (P = 0.058), tumor size (P = 0.001), tumor location (central versus peripheral) (P = 0.015), and multifocality (P = 0.001). Distant dissemination, seeding during surgery, residual disease and new growth are the mechanisms responsible for cancer relapse. Large central lesions, long warm ischemia time and multifocality were significant predictors of oncological failure.


Surgical treatment of trigonocephaly: technique and long-term results in 48 cases.
The authors present their experience in the surgical treatment of metopic synostosis by orbital bandeau remodeling and frontal bone rotation. The pitfalls and advantages of the surgical technique are discussed, along with the long-term clinical results in 48 consecutive cases. Forty-eight consecutive patients in whom trigonocephaly was diagnosed between 1990 and 2009 were treated with frontal bone rotation and frontoorbital bandeau remodeling. Of these patients, 38 (79%) were boys and 10 (21%) were girls. The age at the time of surgical treatment ranged between 4 and 42 months (mean ± SD 11.4 ± 8.7 months). The average follow-up period was 5.5 ± 4.2 years (range 5 months-19 years). The preoperative and latest postoperative photographs of the patients were evaluated for the following features: 1) shape of the forehead; 2) hypotelorism; and 3) temporal depression. Scores of 0, 1, or 2 were assigned for each item: 0 was normal, 1 meant moderate deformity, and 2 denoted severe deformity. In the early postoperative period, no complications were documented. The average hospitalization period was 4 days. Follow-up radiographs or 3D CT scans were obtained at regular intervals. The mean preoperative scores for the evaluated items were 1.38 ± 0.49 for the shape of the forehead, 1.33 ± 0.48 for hypotelorism, and 1.7 ± 0.46 for the temporal depression. The mean postoperative scores were 0.06 ± 0.24 for the shape of the forehead, 0.21 ± 0.4 for hypotelorism, and 0.67 ± 0.48 for the temporal depression. Overall, the total preoperative score dropped from 4.4 to 0.93 postoperatively (p < 0.05). All the patients were contented with the cosmetic results. Early detection and treatment of metopic suture synostosis has a significant, favorable influence on the outcomes. Good understanding of the structural abnormality and the pathophysiological mechanisms of the possible complications is very important for performing proper surgical reconstruction.


SV40 large tumor antigen nuclear import is regulated by the double-stranded DNA-dependent protein kinase site (serine 120) flanking the nuclear localization sequence.
Nuclear localization sequence (NLS)-dependent nuclear import of SV40 large tumor antigen (T-Ag) fusion proteins is regulated by phosphorylation sites for casein kinase II (CKII) and the cyclin-dependent kinase Cdc2 amino-terminal to the NLS (amino acids 126-132). Between the T-Ag CKII and Cdc2 sites is a site (Ser120) for the double-stranded DNA-dependent protein kinase (dsDNA-PK), which we show here for the first time to play a role in regulating T-Ag nuclear import. We replaced Ser120 by aspartic acid or alanine using site-directed mutagenesis and assessed the effects on nuclear transport kinetics both in vivo (microinjected cells) and in vitro (mechanically perforated cells) in HTC rat hepatoma cells. Maximal nuclear accumulation of the Asp120 and Ala120 protein derivatives was approximately 40% and 70% reduced in vivo, respectively, compared with that of the wild type protein, and similarly reduced in vitro, although to a lesser extent. This implies that the dsDNA-PK site regulates the maximal level of nuclear accumulation, normally functioning to enhance T-Ag nuclear transport; the higher accumulation of the Asp120 protein compared with the Ala120 protein indicates that negative charge at the dsDNA-PK site is mechanistically important in regulating nuclear import. The Asp120 protein accumulated in the nucleus at a faster rate than the wild type protein, implying that phosphorylation at Ser120 may also regulate the nuclear import rate. CKII phosphorylation of the Asp120 protein in cytosol or by purified CKII was approximately 30% higher than that of the Ser120 and Ala120 proteins, while negative charge at the CKII site increased dsDNA-PK phosphorylation of Ser120 by approximately 80% compared with wild type, implying physical and functional interactions between the two phosphorylation sites. Quantitation of NLS recognition by the importin 58/97 subunits using an enzyme-linked immunosorbent assay indicated that while the Ala120 protein derivative had a binding affinity very similar to that of wild type, the Asp120 derivative showed 40% higher affinity. In vitro CKII phosphorylation increased importin binding by about 30% in all cases. These results imply that negative charge at the dsDNA-PK site may enhance nuclear import through increasing both NLS recognition by importin subunits, and phosphorylation at the CKII site, which itself also facilitates NLS recognition by importin 58/97.


Impression on observing psychic surgery and healing in Brazil which appear to incorporate (+) qi gong energy & the use of acupuncture points.
In December, 1995, the author had the opportunity to observe an elderly psychic healer of East European origin in Sao Paulo, Brazil. This man specialized in cancer treatment by pointing with the fingers of his right hand at his patients, without actually touching them, spending an average of 30 to 40 minutes with each one. The author considered this to be Qi Gong treatment. In March, 1997 the author also observed 2 leading psychic healers in Brazil. One of them, named Rubens Farias, Jr. is a 43 year-old former engineer and computer programmer if European descent, who is commonly known as "Dr. Fritz III" because he is believed to be the spirit of Dr. Adolf Fritz, a German physician who died during World War I, operates through him. The other is "Dr." Hirota, a 53 year old former farmer of Japanese decent who lives near Campinas. About 120 kilometers outside of Sao Paulo and treats large numbers of people daily using indirect and/or hand-on healing techniques. On March 6, 1997, when the author visited "Dr. Fritz III"'s clinic in Sao Paulo with a group of Brazilian physicians, he was informed the about 1,400 patients had come that day. "Dr Fritz III" examined and treated the majority of the patients in less than one minute each, often asking very quick questions and then immediately beginning treatment. Most patients received injections of a dark-brown solution, which, some of the visiting doctors speculated, may be an iodine solution mixed with either alcohol or a local anesthetic. In many patients, he injected this solution near the pathological area or at an acupuncture point near the pathological area. When the needle of the syringe was in the acupuncture pint, he twirled it with his fingers several times and the withdrew it. Minor surgery was performed in about 1/5th of the patients with whom the author observed. Most of the surgical incisions were made on the midline of the tissue over the spine near the pathological area. The clamping of the blood vessels and the closings of the surgical wounds were performed by licensed surgeons or licensed nurses. Major surgery were done by "Dr. Fritz III" who used a rather primitive and unorthodox cutting technique. Except for major surgery, assistance was performed by volunteer nurses, including his wife. After the surgical wounds were closed, gauze band-aids were applied. When the surface of the gauze facing the wound was examined, it showed strong (+) Qi Gong energy according to the Bi-Digital O-Ring Test. Essentially, "Dr. Fritz III"'s treatment consists of acupuncture, injection of iodine with other substances near the pathological area, and (+) Qi Gong energy stored gauze which is applied to the acupuncture point, pathological area, or the site of surgery. "Dr." Hirota is another famous psychic healer whom the author was able to meet and observe in practice while in Brazil. "Dr." Hirota has been practicing for over 20 years. He usually sees patients who come to his clinic in the mornings and he claims to treat 1,000 to 2,000 patients daily between 9 AM and 12 noon. His main treatment also appears to be the application of external Qi Gong energy through the fingers of his right-hand, in combination with Shiatsu Massage and a manual procedure resembling chiropractic manipulation.


Establishment of stably EBV-transformed cell lines from residual clinical blood samples for use in performance evaluation and quality assurance in molecular genetic testing.
Positive control materials for clinical molecular genetic testing applications are currently in critically short supply or non-existent for many genetically based diseases of public health importance. Here we demonstrate that anonymous, residual, clinical blood samples are potential sources of viable lymphocytes for establishing Epstein-Barr virus (EBV)-transformed blood lymphocyte cell lines. We attempted to transform 34 residual blood samples, and analyzed transformation success with respect to sample age, anticoagulant, storage temperature, volume, hemolysis, and patient age and sex. In univariate analysis, sample age was significantly associated with transformation success (P = 0.002). The success rate was 67% (6 of 9) for samples 1 to 7 days old, 38% (3 of 8) for samples 8 to 14 days old and 0% for samples 15 to 21 (0 of 11) days old. When we controlled for sample age in multivariate logistic regression, anticoagulant and storage temperature approached significance (P = 0.070 and 0.087, respectively; samples in acid citrate dextrose (ACD) and refrigerated samples were more likely to transform). Based on these findings, we suggest that samples collected in either ACD or ethylene diamine tetraacetic acid, and up to 14 days old (refrigerated) or 7 days old (stored ambient), are reasonable candidates for EBV transformation. The transformation rate for samples that met these criteria was 63% (10 of 16). Implementation of this process could help alleviate the shortage of positive control materials for clinical molecular genetic testing.


Template-based B₁ inhomogeneity correction in 3T MRI brain studies.
Low noise, high resolution, fast and accurate T₁ maps from MRI images of the brain can be performed using a dual flip angle method. However, B₁ field inhomogeneity, which is particularly problematic at high field strengths (e.g., 3T), limits the ability of the scanner to deliver the prescribed flip angle, introducing errors into the T₁ maps that limit the accuracy of quantitative analyses based on those maps. A dual repetition time method was used for acquiring a B₁ map to correct that inhomogeneity. Additional inaccuracies due to misregistration of the acquired T₁-weighted images were corrected by rigid registration, and the effects of misalignment on the T₁ maps were compared to those of B₁ inhomogeneity in 19 normal subjects. However, since B₁ map acquisition takes up precious scanning time and most retrospective studies do not have B₁ map, we designed a template-based correction strategy. B₁ maps from different subjects were aligned using a twelve-parameter affine registration. Recomputed T₁ maps showed an important improvement with respect to the noncorrected maps: histograms of all corrected maps exhibited two peaks corresponding to white and gray matter tissues, while unimodal histograms were observed in all uncorrected maps because of the inhomogeneity. A method to detect the best nonsubject-specific B₁ correction based on a set of features was designed. The optimum set of weighting factors for those features was computed. The best available B₁ correction was detected in almost all subjects while corrections comparable to the T₁ map corrected using the B₁ map from the same subject were detected in the others.


Eukaryotic translation initiation factor 4E induced progression of primary human mammary epithelial cells along the cancer pathway is associated with targeted translational deregulation of oncogenic drivers and inhibitors.
Pathologic redirection of translational control by constitutive activation of eukaryotic translation initiation factor 4F (eIF4F), the cap-dependent translation initiation apparatus, is an obligatory step in oncogenesis; however, its mechanism remains undefined. Here, we simulate this pro-oncogenic state by overexpressing eIF4E, the rate-limiting component of eIF4F, in primary human mammary epithelial cells (HMECs) and examine the resultant changes in cell biology and gene expression profiles of total and polyribosome-bound mRNA genome wide. Overexpressed eIF4E rescues primary HMECs from telomere-independent growth arrest and disables checkpoints governing S-phase entry as well as apoptosis in HMECs immortalized by telomerase, imparting cells with proliferative and survival autonomy. Although the transcriptional response to increased eIF4E was modest, the translational response was large, selective, and bidirectional. In addition to translational activation of known and novel eIF4E-responsive oncogenic drivers regulating cell growth and survival, our data unveil previously unrecognized cellular defenses including translational activation of tumor suppressors, translational repression of transcripts enriched with miRNA target sites, and translational modulation of genes governing translation itself. These findings provide insight into the proneoplastic and compensatory mechanisms embedded in the oncogenic translational program. They support a model whereby deregulated eIF4E moves human epithelial cells along the cancer pathway by profoundly altering ribosomal recruitment to cancer-related transcripts, and eIF4E-modified cells counter these potentially oncogenic alterations with a compensatory translational mechanism that mitigates acquisition of malignancy.


Antimicrobial Effect of Simira ecuadorensis Extracts and Their Impact on Improving Shelf Life in Chicken and Fish Products.
The objective of this work was to evaluate the antimicrobial potential of different extracts of Simira ecuadorensis, a characteristic plant of Ecuador, and to validate its potential as a food preservative. Four extracts referred to as ethanol, ethanol-water (50:50 v/v), spray-dried, and freeze-dried were obtained under different processes. Initially, their antimicrobial activities were evaluated against a wide group of microorganisms consisting of 20 pathogenic and spoilage microbial strains found in foods through the agar diffusion method. Then, the extracts with the best yields and antimicrobial properties against microorganisms of greatest interest were selected to determine their effect on model foods preserved under normal commercial conditions through challenge tests. Spray-dried and ethanol-water extracts were tested for their ability to inhibit C. jejuni in chicken model products, where is a common pathogen and Shew. putrefaciens in fish model products as it is a spoilage microorganism frequently found in fish. One solid and one liquid were chosen as model foods: burger and broth, respectively. Campylobacter jejuni and Shewanella putrefaciens were effectively inhibited by the four extracts with minimum inhibitory concentration (MIC) of 80 mg/mL. Bacillus cereus, Yersinia enterocolitica, Clostridium perfringens, and Leuconostoc mesenteroides were also inhibited by ethanolic extract. The ethanol-water extract showed greater antimicrobial activity in fish products, whereas spray-dried extract had low growth inhibition of C. jejuni in chicken burgers; however, it was quite effective on C. jejuni in broth. The spray-dried extract significantly decreased the pH of the chicken burgers, while the ethanolic extract had a slight impact on the pH of the fish burgers. The presence of antibacterial effects revealed that the S. ecuadorensis extracts could be potentially used in food preservation and as a natural antimicrobial.


Pulsatile hyperglucagonemia fails to increase hepatic glucose production in normal man.
To study the metabolic effects of pulsatile glucagon administration, six male volunteers were submitted to a 260-min glucose-controlled glucose intravenous infusion using the Biostator. The endogenous secretion of the pancreatic hormones was inhibited by somatostatin (100 micrograms X h-1), basal insulin secretion was replaced by a continuous insulin infusion (0.2 mU X kg-1 X min-1), and glucagon was infused intravenously in two conditions at random: either continuously (125 ng X min-1) or intermittently (812.5 ng X min-1, with a switching on/off length of 2/11 min). Blood glucose levels and glucose infusion rate were monitored continuously by the Biostator, and classical methodology using a D-[3-3H]glucose infusion allowed us to study glucose turnover. While basal plasma glucagon levels were similar in both conditions (122 +/- 31 vs. 115 +/- 18 pg X ml-1), they plateaued at 189 +/- 38 pg X ml-1 during continuous infusion and varied between 95 and 501 pg X ml-1 during pulsatile infusion. When compared with continuous administration, pulsatile glucagon infusion initially induced a similar increase in endogenous (hepatic) glucose production and blood glucose, did not prevent the so-called "evanescent" effect of glucagon on blood glucose, and after 3 h tended to reduce rather than increase hepatic glucose production. In conclusion, in vivo pulsatile hyperglucagonemia in normal man fails to increase hepatic glucose production.


Triptych on Stress, Anxiety, and Loneliness.
Three digital drawings invite interpretations of a cornered figure's orientations to space, color, and its own boundaries and self. Little variation among lines and spaces, their colors and shades, and the figure's position suggest the representational importance of each to discerning meaning about how we forge connection.


Comparison of cryoprotective effects of lycopene and cysteamine in different cryoprotectants on bull semen and fertility results.
The objectives of this study were to compare glycerol and ethylene glycol at different concentrations as cryoprotectants and lycopene or cysteamine (with/without) as antioxidants in Tris extender for bull semen. Twenty-four ejaculates were obtained from three bulls. Each ejaculate was split into four equal aliquots and diluted using both of the Tris extenders with glycerol (5% or 7%) or ethylene glycol (3% or 5%). After that, each extenders were split into three equal aliquots and added using both of the cysteamine 5 mm or lycopene 500 μg/ml, and control (without additives). The addition of 7% glycerol with cysteamine, 5% ethylene glycol with cysteamine and 3% ethylene glycol with cysteamine groups gave the lowest CASA motility than the other groups. However, 7% glycerol and 7% glycerol with lycopene resulted in a better rate of CASA progressive motility compared with that of other groups. Generally, all the lycopene groups signed better protective effects on acrosome and total morphology than the other groups. Glycerol 7% and 3% ethylene glycol with lycopene groups yielded to slight higher percentages of membrane integrity assessed by HOST than that of the other groups, but 7% glycerol with cysteamine and 3% ethylene glycol with cysteamine showed the worst percentages of membrane integrity. Glycerol 7% and 5% glycerol with lycopene gave rise to a higher value of VAP, VSL and VCL compared with that of the other groups. On the contrary, adding to 5% glycerol with cysteamine showed negative effect for VAP, VSL, VCL and ALH values. All cryoprotectant groups with lycopene decreased chromatin damage than the other groups. Ethylene glycol 3% led to lower non-return rates of inseminated cows. However, this result was not considered to be statistically important.


Expression of small cytoplasmic transcripts of the rat identifier element in vivo and in cultured cells.
We examined the level of expression of small RNA transcripts hybridizing to a rodent repetitive DNA element, the identifier (ID) sequence, in a variety of cell types in vivo and in cultured mammalian cells. A 160-nucleotide (160n) cytoplasmic poly(A)+ RNA (BC1) appeared in late embryonic and early postnatal rat brain development, was enriched in the cerebral cortex, and appeared to be restricted to neural tissue and the anterior pituitary gland. A 110n RNA (BC2) was specifically enriched in brain, especially the postnatal cortex, but was detectable at low levels in peripheral tissues. A third, related 75n poly(A)- RNA (T3) was found in rat brain and at lower levels in peripheral tissues but was very abundant in the testes. The BC RNAs were found in a variety of rat cell lines, and their level of expression was dependent upon cell culture conditions. A rat ID probe detected BC-like RNAs in mouse brain but not liver and detected a 200n RNA in monkey brain but not liver at lower hybridization stringencies. These RNAs were expressed by mouse and primate cell lines. Thus, tissue-specific expression of small ID-sequence-related transcripts is conserved among mammals, but the tight regulation found in vivo is lost by cells in culture.