Education and Protein Supplementation Improve Nutritional Biomarkers among Hypoalbuminemic Peritoneal Dialysis Patients: A Quasi-Experimental Design.
Protein-energy wasting is prevalent in peritoneal dialysis patients, which causes a heavy burden for individuals and healthcare systems. We aimed to investigate the effect of nutritional education, and/or protein supplementation on nutritional biomarkers in hypoalbuminemic peritoneal dialysis patients. A quasi-experimental study was conducted in two dialysis centers at Taipei Tzu Chi Hospital and Shin Kong Wu Ho-Su Memorial Hospital. Patients were allocated in three groups including control (n = 12), milk protein (n = 21) and soy protein (n = 20). All patients received dietary guidelines from dietitians and completed 3-day dietary records during monthly visits for consecutive three months. Nutrients were analyzed using Nutritionist Professional software. Blood urea nitrogen (BUN), creatinine, albumin, total protein, hemoglobin, serum calcium, phosphorus, sodium, and potassium were assessed monthly. Total cholesterol and triglycerides were measured every three months. After three-month intervention, protein intake (percent of total calories), and serum albumin were significantly increased in three groups. Protein, phosphorus intake, and BUN were increased in two intervention groups. Total serum protein increased in control and milk protein groups, and creatinine increased the control group. Serum phosphorus was not significantly changed. Nutritional education alone, or combined with protein supplementation, significantly improve protein intake, and nutritional status by increasing serum albumin, but not serum phosphorus in hypoalbuminemic peritoneal dialysis patients.


Impact of modified quality control checklist on protocol adherence and outcomes in a post-surgical Intensive Care Unit.
Quality improvement (QI) is the sum of all activities that create desired changes in the quality. An effective QI system results in a stepwise increase in quality of care. The efficiency of any health-care unit is judged by its quality indicators. We aimed to evaluate the impact of QI initiatives on outcomes in a surgical Intensive Care Unit (ICU). This was an observational study carried out using a compliance checklist, developed from the combination of the World Health Organization surgery checklist and Society for Healthcare Epidemiology of America guidelines for the prevention of infections. A total of 170 patients were prospectively evaluated for adherence to the checklist and occurrence of infections. This was compared with a random retrospective analysis of 170 patients who had undergone similar surgeries in the previous 3 months. Introduction and supervised documentation of comprehensive checklist brought out significant improvement in the documentation of quality indicators (98% vs. 32%) in the prospective samples. There was no difference in mortality, health-care-related infection rates or length of ICU stay. The introduction of comprehensive surgical checklist improved documentation of parameters for quality control but did not decrease the rates of infection in comparison to the control sample.


Potassium ion controlled switching of intra- to intermolecular electron transfer in crown ether appended free-base porphyrin-fullerene donor-acceptor systems.
Photoinduced electron transfer in intramolecularly interacting free-base porphyrin bearing one or four 18-crown-6 ether units at different positions of the porphyrin macrocycle periphery and pristine fullerene was investigated in polar benzonitrile and nonpolar o-dichlorobenzene and toluene solvents. Owing to the presence of two modes of binding, stable dyads were obtained in which the binding constants, K, were found to range between 4.2 x 10(3) and 10.4 x 10(3) M(-1) from fluorescence quenching data depending upon the location and number of crown ether entities on the porphyrin macrocycle and the solvent. Computational studies using the B3LYP/3-21G() method were employed to arrive at the geometry and electronic structure of the intramolecular dyads. The energetics of the redox states of the dyads were established from cyclic voltammetric studies. Under the intramolecular conditions, both the steady-state and time-resolved emission studies revealed efficient quenching of the singlet excited free-base porphyrin in these dyads, and the measured rates of charge separation, k(CS), were found to be in the 10(8)-10(9) s(-1) range. Nanosecond transient absorption studies were performed to characterize the electron-transfer products and to evaluate the charge-recombination rates. Shifting of the electron-transfer pathway from the intra- to intermolecular route was achieved by complexing potassium ions to the crown ether cavity(ies) in benzonitrile. This cation complexation weakened the intramolecular interactions between fullerene and the crown ether appended free-base porphyrin supramolecules, and under these conditions, intermolecular type interactions were mainly observed. Reversible inter- to intramolecular electron transfer was also accomplished by extracting the potassium ions of the complex with the addition of 18-crown-6. The present study nicely demonstrates the application of supramolecular methodology to control the excited-state electron-transfer path in donor-acceptor dyads.


Dexamethasone with bupivacaine increases duration of analgesia in ultrasound-guided interscalene brachial plexus blockade.
Dexamethasone has been shown to prolong the duration of postoperative analgesia when given as an adjunct for peripheral nerve blocks. However, it has not been evaluated when given in conjunction with bupivacaine and clonidine to provide blockade of the brachial plexus at the interscalene level. The purpose of this investigation was to determine whether the addition of dexamethasone to interscalene brachial plexus block would prolong the duration of sensory analgesia in a group of patients undergoing outpatient shoulder arthroscopy. This prospective, randomized, double-blind investigation was performed on 88 individuals undergoing shoulder arthroscopy. Patients received interscalene brachial plexus block using 20 ml of bupivacaine 5 mg ml(-1) with 1: 200,000 epinephrine and clonidine 75 microg. Patients were randomly assigned to receive either dexamethasone 8 mg or 0.9% NaCl as an adjuvant to the mixture. After discharge, patients recorded pain scores and analgesic consumption in a diary and estimated the time at which they perceived that the sensory block from the interscalene brachial plexus block resolved. This was based on pain, recovery of sensation and strength in the arm. Variables measured included demographics, timed pain intensity measurements, postoperative analgesic consumption, duration of analgesia and patient satisfaction. Dexamethasone prolonged median sensory (1457 vs. 833 min, P < 0.0001) and motor (1374 vs. 827 min, P < 0.0001) blockade compared with the control. At 24 h, the dexamethasone group had lower median verbal analogue scale scores compared with control (3.0 vs. 6.0). At 48 h, the two groups had similar median pain scores (4.0 vs. 5.0, dexamethasone vs. control, respectively). The opioid requirement in oxycodone equivalency was lower in the dexamethasone group than in the control group for the first 24 h, and similar thereafter. Median patient satisfaction scores were not significantly different between the two groups at 48 h (9.5 vs. 8.0, dexamethasone vs. control, respectively). The addition of dexamethasone to a bupivacaine-epinephrine-clonidine interscalene block prolongs sensory block and reduces opioid use.


The helical domains of the stem region of dengue virus envelope protein are involved in both virus assembly and entry.
The envelope (E) of dengue virus (DENV) is a determinant of tropism and virulence. At the C terminus of E protein, there is a stem region containing two amphipathic α-helical domains (EH1 and EH2) and a stretch of conserved sequences in between. The crystal structure of E protein at the postfusion state suggested the involvement of the stem during the fusion; however, the critical domains or residues involved remain unknown. Site-directed mutagenesis was carried out to replace each of the stem residues at the hydrophobic face with an alanine or proline in a DENV serotype 4 (DENV4) precursor membrane (prM)/E expression construct. Most of the 15 proline mutations at either EH1 or EH2 severely affected the assembly of virus-like particles (VLPs). Radioimmunoprecipitation and membrane flotation assays revealed that EH1 mutations primarily affect prM-E heterodimerization and EH2 mutations affect the membrane binding of the stem. Introducing four proline mutations at either EH1 or EH2 into a DENV2 replicon packaging system greatly affects assembly and entry. Moreover, introducing these mutations into a DENV2 infectious clone confirmed the impairment in assembly and infectivity. Sequencing analysis of adaptive mutations in passage 5 viruses revealed a change to a leucine or wild-type residue at the original site, suggesting the importance of maintaining the helical structure. Collectively, these findings suggest that the EH1 and EH2 domains are involved in both assembly and entry steps of the DENV replication cycle; this feature, together with the high degree of sequence conservation, suggests that the stem region is a potential target of antiviral strategies.


Small for size liver remnant following resection: prevention and management.
In the latest decades an important change was registered in liver surgery, however the management of liver cirrhosis or small size hepatic remnant still remains a challenge. Currently post-hepatectomy liver failure (PLF) is the major cause of death after liver resection often associated with sepsis and ischemia-reperfusion injury (IRI). ''Small-for-size'' syndrome (SFSS) and PFL have similar mechanism presenting reduction of liver mass and portal hyper flow beyond a certain threshold. Few methods are described to prevent both syndromes, in the preoperative, perioperative and postoperative stages. Additionally to portal vein embolization (PVE), radiological examinations (mainly CT and/or MRI), and more recently 3D computed tomography are fundamental to quantify the liver volume (LV) at a preoperative stage. During surgery, in order to limit parenchymal damage and optimize regenerative capacity, some hepatoprotective measures may be employed, among them: intermittent portal clamping and hypothermic liver preservation. Regarding the treatment, since PLF is a quite complex disease, it is required a multi-disciplinary approach, where it management must be undertaken in conjunction with critical care, hepatology, microbiology and radiology services. The size of the liver cannot be considered the main variable in the development of liver dysfunction after extended hepatectomies. Additional characteristics should be taken into account, such as: the future liver remnant; the portal blood flow and pressure and the exploration of the potential effects of regeneration preconditioning are all promising strategies that could help to expand the indications and increase the safety of liver surgery.


Pediatric Death After Withdrawal of Life-Sustaining Therapies: A Scoping Review.
Evaluate literature on the dying process in children after withdrawal of life sustaining measures (WLSM) in the PICU. We focused on the physiology of dying, prediction of time to death, impact of time to death, and uncertainty of the dying process on families, healthcare workers, and organ donation. MEDLINE, Embase, Cochrane Central Register of Controlled Trials, PsycINFO, CINAHL, and Web of Science. We included studies that discussed the dying process after WLSM in the PICU, with no date or study type restrictions. We excluded studies focused exclusively on adult or neonatal populations, children outside the PICU, or on organ donation or adult/pediatric studies where pediatric data could not be isolated. Inductive qualitative content analysis was performed. Six thousand two hundred twenty-five studies were screened and 24 included. Results were grouped into four categories: dying process, perspectives of healthcare professionals and family, WLSM and organ donation, and recommendations for future research. Few tools exist to predict time to death after WLSM in children. Most deaths after WLSM occur within 1 hour and during this process, healthcare providers must offer support to families regarding logistics, medications, and expectations. Providers describe the unpredictability of the dying process as emotionally challenging and stressful for family members and staff; however, no reports of families discussing the impact of time to death prediction were found. The unpredictability of death after WLSM makes families less likely to pursue donation. Future research priorities include developing death prediction tools of tools, provider and parental decision-making, and interventions to improve end-of-life care. The dying process in children is poorly understood and understudied. This knowledge gap leaves families in a vulnerable position and the clinical team without the necessary tools to support patients, families, or themselves. Improving time to death prediction after WLSM may improve care provision and enable identification of potential organ donors.


Surface plasmon resonance sensor based on polarization interferometry and angle modulation.
A surface plasmon resonance (SPR) sensing technique based on polarization interferometry and angle modulation is presented. Its sensitivity is not a direct function of variation of reflection intensity, nor of phase shift. Rather, it is a function of the complex reflection coefficient. A three times standard deviation detection limit of 5.1 x 10(-7) refractive index units in a 2 Hz bandwidth is obtained with our experimental setup. A theoretical analysis shows that this technique can provide a wide linear measurement range. Moreover, the sensitivity is insensitive to the thickness of gold films over approximately 5 nm. This SPR sensing technique is suitable for physical, chemical, and biological research.


Encapsulated environment.
In many occupational settings, clothing must be worn to protect individuals from hazards in their work environment. However, personal protective clothing (PPC) restricts heat exchange with the environment due to high thermal resistance and low water vapor permeability. As a consequence, individuals who wear PPC often work in uncompensable heat stress conditions where body heat storage continues to rise and the risk of heat injury is greatly enhanced. Tolerance time while wearing PPC is influenced by three factors: (i) initial core temperature (Tc), affected by heat acclimation, precooling, hydration, aerobic fitness, circadian rhythm, and menstrual cycle (ii) Tc tolerated at exhaustion, influenced by state of encapsulation, hydration, and aerobic fitness; and (iii) the rate of increase in Tc from beginning to end of the heat-stress exposure, which is dependent on the clothing characteristics, thermal environment, work rate, and individual factors like body composition and economy of movement. Methods to reduce heat strain in PPC include increasing clothing permeability for air, adjusting pacing strategy, including work/rest schedules, physical training, and cooling interventions, although the additional weight and bulk of some personal cooling systems offset their intended advantage. Individuals with low body fatness who perform regular aerobic exercise have tolerance times in PPC that exceed those of their sedentary counterparts by as much as 100% due to lower resting Tc, the higher Tc tolerated at exhaustion and a slower increase in Tc during exercise. However, questions remain about the importance of activity levels, exercise intensity, cold water ingestion, and plasma volume expansion for thermotolerance.


Phylogenetic analysis based on 18S rRNA gene and matK gene sequences of Panax vietnamensis and five related species.
Panax vietnamensis was discovered recently in Vietnam. Its bamboo-like rhizomes, called Vietnamese Ginseng, have attracted considerable attention because of their specific pharmacological activities. In order to define the taxonomic position of this new species and include it in the molecular authentication of Ginseng drugs, the 18S ribosomal RNA gene and matK gene sequences of P. vietnamensis were determined and compared with those of its related taxa, P. japonicus var. major and P. pseudo-ginseng subsp. himalaicus, besides previously reported P. ginseng, P. japonicus and P. quinquefolius. The 18S rRNA gene sequences were found to be 1809 bps in length. The sequence of P. vietnamensis was identical to that of P. quinquefolius, and presented one base substitution from those of both P. japonicus var. major and P. pseudo-ginseng subsp. himalaicus. The matK gene sequences of 6 taxa were found to be 1509 bps in length. The sequence of P. vietnamensis differed from those of P. japonicus var. major, P. pseudo-ginseng subsp. himalaicus, P. ginseng, P. japonicus and P. quinquefolius at 4, 5, 9, 9 and 10 nucleotide positions, respectively. The phylogenetic tree reconstructed by the combined 18S rRNA-matK gene analysis using the maximum parsimony method showed that P. vietnamensis was sympatric with other Panax species and had a close relationship with P. japonicus var. major and P. pseudo-ginseng subsp. himalaicus.


Perceptions of Contextual Stressors in Physical Education. A Qualitative Case Study.
Background: Daily stressors have a significant impact on students' educational outcomes. However, research on students perceived and common contextual stressors in physical education (PE) lessons is limited. Purpose: To identify potential contextual stressors in PE contexts and what students perceive as stressors. Participants: Ninth-grade students (age 14-15) and their PE teachers recruited from three classes in one lower secondary school in Norway. Research Design: This qualitative case study used data generated from descriptive field notes from participant observations in PE lessons, formal interviews and informal conversations with PE teachers, focus group and individual interviews with students, and a supplementary survey using the TurningPoint student response system. Conversations were transcribed verbatim and analyzed using reflexive thematic analysis (Braun and Clarke, 2006; Tolmie et al., 2011; Braun et al., 2019) and the NVivo 12 Pro analysis software. The survey was analyzed using IBM SPSS Statistics 21. Findings: This study supports and expands previous research exploring students' stressors in PE and highlights the volume and variety of potential stressors in PE contexts. The findings shed light on certain similarities and differences that may exist between students of different genders and grades and with different past physical activity experiences. In the present study, spectators, in addition to difficult tasks and low self-efficacy, seemed particularly stressful for girls. This article presents nuances revealed by various qualitative approaches and a supplementary survey. Conclusion: Students in this study experience a multitude of stressors during PE lessons. These include stressors in the teaching, physical, and social environments, as well as personal factors. The stressors experienced depend on the situation, the lesson content, the parties involved, students' past experiences, and their appraisal of these stressors. In our sample, girls seemed to be more vulnerable to contextual stressors in PE than boys.


Molecular characterisation of hepatitis B virus in HIV-1 subtype C infected patients in Botswana.
Hepatitis B virus (HBV) is a major global health problem especially in sub-Saharan Africa and in East Asia. Ten hepatitis B virus genotypes have been described that differ by geographic distribution, disease progression, and response to treatment. Escape mutations within the surface open reading frame (ORF) affect HBV antigenicity leading to failures in diagnosis, vaccine and hepatitis B immunoglobulin therapy. However, the molecular characteristics of HBV in Botswana, a highly endemic country, are unknown. We describe the molecular characteristics of HBV and prevalence of escape mutants among HIV/HBV coinfected individuals Botswana. DNA was extracted from archived plasma samples from 81 HIV/HBV co-infected participants from various clinical studies at the Botswana Harvard AIDS Institute Partnership. A 415 base pair (bp) fragment of the polymerase gene was amplified by semi-nested PCR. In a subset of samples, a 2100 bp fragment was amplified. The PCR product was genotyped using Big Dye sequencing chemistry and the sequences were analysed for genotypes and mutations. Of the 81 samples included, 70 (86 %) samples were successfully genotyped. Genotype A was found in 56 (80 %) participants, D in 13 (18.6 %), and 1 (1.4 %) was genotype E. Escape mutations previously linked with failure of diagnosis or escaping active vaccination and passive immunoglobulin therapy were detected in 12 (17.1 %) participants at positions 100, 119, 122, 123, 124, 126, 129, 130, 133, 134 and 140 of the S ORF. Genotypes and escape mutations were not significantly associated with aspartate aminotransferase (AST), alanine aminotransferase (ALT) and AST platelet ratio index (APRI). Genotypes A, D and E were found in this cohort of HIV coinfected patients in Botswana, consistent with the findings from the sub-Saharan Africa region. Some escape mutations which have previously been associated with diagnosis failure, escaping vaccine and immunoglobulin therapy were also observed and are important in guiding future policies related to vaccine implementation, therapeutic guidelines, and diagnostic guidelines. They are also important for identifying patients who are at an increased risk of disease progression and to choose optimal therapy. Future research should focus on determining the clinical significance of the different HBV genotypes and mutations found in this population.


Disclosure of medical errors: what factors influence how patients respond?
Disclosure of medical errors is encouraged, but research on how patients respond to specific practices is limited. This study sought to determine whether full disclosure, an existing positive physician-patient relationship, an offer to waive associated costs, and the severity of the clinical outcome influenced patients' responses to medical errors. Four hundred and seven health plan members participated in a randomized experiment in which they viewed video depictions of medical error and disclosure. Subjects were randomly assigned to experimental condition. Conditions varied in type of medication error, level of disclosure, reference to a prior positive physician-patient relationship, an offer to waive costs, and clinical outcome. Self-reported likelihood of changing physicians and of seeking legal advice; satisfaction, trust, and emotional response. Nondisclosure increased the likelihood of changing physicians, and reduced satisfaction and trust in both error conditions. Nondisclosure increased the likelihood of seeking legal advice and was associated with a more negative emotional response in the missed allergy error condition, but did not have a statistically significant impact on seeking legal advice or emotional response in the monitoring error condition. Neither the existence of a positive relationship nor an offer to waive costs had a statistically significant impact. This study provides evidence that full disclosure is likely to have a positive effect or no effect on how patients respond to medical errors. The clinical outcome also influences patients' responses. The impact of an existing positive physician-patient relationship, or of waiving costs associated with the error remains uncertain.


Mood state and quality of sleep in cancer pain patients: a comparison to chronic daily headache.
Cancer pain is commonly believed to be a unique type of pain and dissimilar to noncancer pain; however, only limited research efforts have been directed at examining this belief. The aim of this study was to explore whether patients with chronic daily headache (CDH) and patients with chronic cancer pain (CCP) present with different pain, mood, and sleep quality profiles. Forty-seven patients diagnosed with CDH were matched by age and gender with 47 patients with CCP. The research instruments included the Brief Pain Inventory-Chinese version, the Profile of Mood States Short Form, and the Pittsburgh Sleep Quality Index-Taiwan Form (PSQI-T). Results revealed that there was no difference in pain intensity between the patients with CDH and those with CCP; however, the CCP group reported significantly higher mean levels of pain interference with daily life than did the CDH group. These two groups did not differ on the Total Mood Disturbance score; however, the CCP group reported significantly lower mean levels of vigor than did the CDH group. Moreover, there was no difference on the PSQI-T total score between these two groups; however, the CDH group reported higher mean scores of sleep disturbance, higher mean scores of use of sleep medications, lower mean scores of sleep efficiency, and lower mean scores of daytime dysfunction than did the CCP group. Despite some differences between these two groups, pain, mood, and sleep quality profiles in these two types of pain groups are similar.


Turnip yellow mosaic virus: transfer RNA mimicry, chloroplasts and a C-rich genome.
SUMMARY Taxonomy: Turnip yellow mosaic virus (TYMV) is the type species of the genus Tymovirus, family Tymoviridae. TYMV is a positive strand RNA virus of the alphavirus-like supergroup. Physical properties: Virions are non-enveloped 28-nm T = 3 icosahedrons composed of a single 20-kDa coat protein that is clustered in 20 hexameric and 12 pentameric subunits. Infectious particles and empty capsids coexist in infected tissue. The genomic RNA is 6.3 kb long, with a 5'(m7)GpppG cap and a 3' untranslated region ending in a tRNA-like structure to which valine can be covalently added. The genome has a distinctive skewed C-rich, G-poor composition (39% C, 17% G). Viral proteins: Two proteins, whose open reading frames extensively overlap, are translated from the genomic RNA. p206, which contains sequences indicative of RNA capping, NTPase/helicase and polymerase activities, is the only viral protein that is necessary for genome replication in single cells. It is produced as a polyprotein and self-cleaved to yield 141- and 66-kDa proteins. p69 is required for virus movement within the plant and is also a suppressor of gene silencing. The coat protein is expressed from the single subgenomic RNA. Hosts and symptoms: TYMV has a narrow host range almost completely restricted to the Cruciferae. Experimental host species are Brassica pekinensis (Chinese cabbage) or B. rapa (turnip), in which diffuse chlorotic local lesions and systemic yellow mosaic symptoms appear. Arabidopsis thaliana can also be used. Clumping of chloroplasts and the accumulation of vesicular invaginations of the chloroplast outer membranes are distinctive cytopathological symptoms. High yields of virus are produced in all leaf tissues, and the virus is readily transmissible by mechanical inoculation. Localized transmission by flea beetles may occur in the field.


Neuropathic pain management in children.
There are difficulties in assessing, managing, and evaluating neuropathic pain in dying children, particularly those with neurological impairment. Neuropathic pain in children often presents differently to how it presents in the adult population. Comprehensive assessment as well as pharmacological and non-pharmacological interventions are crucial to its successful management and frequently require input from an interdisciplinary team. Notwithstanding the need for further research, this paper brings together research papers, reviews, and clinical guidelines to present an exploration of existing evidence regarding care for children with neuropathic pain and their families.


Diverse requirements for Notch signalling in mammals.
The Notch signalling pathway has a central role in a wide variety of developmental processes and it is not therefore surprising that mutations in components of this pathway can cause dramatic human genetic disorders. One developmental process in which the Notch pathway is involved at multiple levels is somitogenesis, the mechanism by which the embryo is divided into segments that ultimately form structures such as the axial skeleton and skeletal muscle of the trunk. We are investigating the human genetic disorder spondylocostal dysplasia (SCD), which is a group of malsegmentation syndromes that occur when this process is disrupted. Mutations in the Notch ligand DELTA-LIKE 3 (DLL3) are responsible for cases of autosomal recessive SCD type I (SCDO1), and we are using information derived from these mutations to study the structure of the DLL3 protein. To aid in elucidation of the underlying developmental defect in SCDO1, we have generated a mouse model by targeted deletion of the Dll3 gene (Dunwoodie et al., 2002). These mice show segmentation defects similar to those seen in SCDO1. In addition, these mice have a distinct set of neural defects that may be useful in future neurological assessment of affected individuals. Finally, since not all cases of SCD are due to mutation of DLL3, we are investigating various genes to find other candidates involved in this genetic disease.


Gene regulatory network analysis predicts cooperating transcription factor regulons required for FLT3-ITD+ AML growth.
Acute myeloid leukemia (AML) is a heterogeneous disease caused by different mutations. Previously, we showed that each mutational subtype develops its specific gene regulatory network (GRN) with transcription factors interacting within multiple gene modules, many of which are transcription factor genes themselves. Here, we hypothesize that highly connected nodes within such networks comprise crucial regulators of AML maintenance. We test this hypothesis using FLT3-ITD-mutated AML as a model and conduct an shRNA drop-out screen informed by this analysis. We show that AML-specific GRNs predict crucial regulatory modules required for AML growth. Furthermore, our work shows that all modules are highly connected and regulate each other. The careful multi-omic analysis of the role of one (RUNX1) module by shRNA and chemical inhibition shows that this transcription factor and its target genes stabilize the GRN of FLT3-ITD+ AML and that its removal leads to GRN collapse and cell death.


Potential drug-drug interactions in infant, child, and adolescent patients in children's hospitals.
Hospitalized infants, children, and adolescents are typically exposed to numerous distinct medications during inpatient admissions, increasing their risk of potential drug-drug interactions (PDDIs). We assessed the prevalence and characteristics of PDDI exposure of pediatric patients treated in children's hospitals. This retrospective cohort study included patients <21 years old hospitalized in children's hospitals throughout the United States. PDDIs were identified by using the MicroMedex DRUG-REAX system. We calculated the patients exposed to PDDIs, stratified according to the seriousness of the interaction; daily and cumulative counts of PDDI exposures; and characterization of the cited potential adverse effects. Of 498 956 hospitalizations in 2011, 49% were associated with ≥1 PDDI, with a "contraindicated" PDDI occurring in 5% of all hospitalizations, a "major" PDDI present in 41%, a "moderate" PDDI in 28%, and a "minor" PDDI in 11%. Opioids were involved in 25% of all PDDIs, followed by antiinfective agents (17%), neurologic agents (15%), gastrointestinal agents (13%), and cardiovascular agents (13%). One-half of all PDDI exposures were due to specific drug pairs occurring in ≤3% of patients per hospital day. The most common potential adverse drug events included additive respiratory depression (in 21% of PDDIs), bleeding risk (5%), QT interval prolongation (4%), reduced iron absorption/availability (4%), central nervous system depression (4%), hyperkalemia (3%), and altered diuretic effectiveness (3%). Exposure to PDDIs is common among hospitalized children. Empirical data are needed to determine the probability and magnitude of the actual harm for each specific PDDI, particularly for less common drug pairs.


Biochemical Network Stochastic Simulator (BioNetS): software for stochastic modeling of biochemical networks.
Intrinsic fluctuations due to the stochastic nature of biochemical reactions can have large effects on the response of biochemical networks. This is particularly true for pathways that involve transcriptional regulation, where generally there are two copies of each gene and the number of messenger RNA (mRNA) molecules can be small. Therefore, there is a need for computational tools for developing and investigating stochastic models of biochemical networks. We have developed the software package Biochemical Network Stochastic Simulator (BioNetS) for efficiently and accurately simulating stochastic models of biochemical networks. BioNetS has a graphical user interface that allows models to be entered in a straightforward manner, and allows the user to specify the type of random variable (discrete or continuous) for each chemical species in the network. The discrete variables are simulated using an efficient implementation of the Gillespie algorithm. For the continuous random variables, BioNetS constructs and numerically solves the appropriate chemical Langevin equations. The software package has been developed to scale efficiently with network size, thereby allowing large systems to be studied. BioNetS runs as a BioSpice agent and can be downloaded from http://www.biospice.org. BioNetS also can be run as a stand alone package. All the required files are accessible from http://x.amath.unc.edu/BioNetS. We have developed BioNetS to be a reliable tool for studying the stochastic dynamics of large biochemical networks. Important features of BioNetS are its ability to handle hybrid models that consist of both continuous and discrete random variables and its ability to model cell growth and division. We have verified the accuracy and efficiency of the numerical methods by considering several test systems.


Structural basis of ubiquitin recognition by translesion synthesis DNA polymerase ι.
Cells have evolved mutagenic bypass mechanisms that prevent stalling of the replication machinery at DNA lesions. This process, translesion DNA synthesis (TLS), involves switching from high-fidelity DNA polymerases to specialized DNA polymerases that replicate through a variety of DNA lesions. In eukaryotes, polymerase switching during TLS is regulated by the DNA damage-triggered monoubiquitylation of PCNA. How the switch operates is unknown, but all TLS polymerases of the so-called Y-family possess PCNA and ubiquitin-binding domains that are important for their function. To gain insight into the structural mechanisms underlying the regulation of TLS by ubiquitylation, we have probed the interaction of ubiquitin with a conserved ubiquitin-binding motif (UBM2) of Y-family polymerase Polι. Using NMR spectroscopy, we have determined the structure of a complex of human Polι UBM2 and ubiquitin, revealing a novel ubiquitin recognition fold consisting of two α-helices separated by a central trans-proline residue conserved in all UBMs. We show that, different from the majority of ubiquitin complexes characterized to date, ubiquitin residue Ile44 only plays a modest role in the association of ubiquitin with Polι UBM2. Instead, binding of UBM2 is centered on the recognition of Leu8 in ubiquitin, which is essential for the interaction.


Workaholism as a risk factor for depressive mood, disabling back pain, and sickness absence.
Although it is understood that work-related factors, including job demands, job control, and workplace support, are associated with workers' health and well-being, the role played by personal characteristics, especially workaholism, has not been fully investigated. This study examined workaholism's associations with psychological ill health, low back pain with disability, and sickness absence among Japanese workers. A cross-sectional Internet survey was conducted using self-administered questionnaires. Data from 3,899 Japanese workers were analyzed. Workaholism was measured using the Dutch Workaholism Scale (DUWAS). Scores were divided into tertiles, where respondents were classified into three groups (high, middle, and low). Depressive mood as a measure of psychological ill health was assessed using the SF-36 mental health subscale, and low back pain using a standardized question. Sickness absence, except that due to physical injuries, was categorized either as absence due to mental health problems or to physical/somatic problems including the common cold. Multiple logistic regression analyses were conducted to examine the association between workaholism and depressive mood, low back pain with disability, and sickness absence, adjusting for demographic characteristics, job demand, job control, and workplace support. Compared to the low workaholism group, the middle and high workaholism groups had significantly higher odds for depressive mood (Odds ratio (OR) = 1.93 and 3.62 for the middle and high groups, respectively), disabling back pain (ORs = 1.36 and 1.77 for the middle and high groups, respectively). Workaholism was more strongly associated with sickness absence due to mental health problems than that for other reasons (ORs = 1.76 vs. 1.21 for the middle group and 3.52 vs. 1.37 for the high groups). Workaholism is significantly associated with poor psychological health, disabling back pain, and sickness absence, particularly from mental health problems. Therefore, workaholism must be considered when addressing well-being of workers.


Kinetics of elimination of urinary metabolites of acrylamide in humans.
Acrylamide (AM), used in the manufacture of polyacrylamide and grouting agents, is produced during the cooking of foods. Workplace exposure to AM can occur through the dermal and inhalation routes. The objective of this study was to define the kinetics of elimination of AM and its metabolites following oral and dermal administration. This is the second part of a study in which metabolites and hemoglobin adducts of AM were determined in people (Fennell et al., 2005, Toxicol. Sci. 85, 447-459). (1,2,3-(13)C(3))AM was administered in an aqueous solution orally (single dose of 0.5, 1.0, or 3.0 mg/kg) or dermally (three daily doses of 3.0 mg/kg) to sterile male volunteers. Urine samples were collected at 0-2, 2-4, 4-8, 8-16, and 16-24 h following administration orally, or at 0-2, 2-4, 4-8, 8-16, and 16-24 h following each of three daily dermal doses. (13)C(3)-AM and its metabolites in urine, (13)C(3)-glycidamide, (13)C(3)-N-acetyl-S-(3-amino-3-oxopropyl)cysteine and its S-oxide, and (13)C(3)-N-acetyl-S-(3-amino-2-hydroxy-3-oxopropyl)cysteine, were quantitated using liquid chromatography-tandem mass spectrometry. The recovered urinary metabolites accounted for 45.6, 49.9, and 39.9% of a 0.5, 1.0, and 3.0 mg/kg oral dose (0-24 h), respectively, and for 4.5% of the dose after 3 mg/kg was administered daily for 3 days dermally (0-4 days). These results indicate that after oral administration AM is rapidly absorbed and eliminated. The half-life estimated for elimination of AM in urine was 3.1-3.5 h. After dermal administration, AM uptake is slow. This study indicated that skin provides a barrier that slows the absorption of AM, and results in limited systemic availability following dermal exposure to AM.


Organ donation: a communitarian approach.
Recently, various suggestions have been made to respond to the increasingly great shortage of organs by paying for them. Because of the undesirable side effects of such approaches (commodification, injustice, and costs), a communitarian approach should be tried first. A communitarian approach to the problem of organ shortage entails changing the moral culture so that members of society will recognize that donating one's organs, once they are no longer of use to the donor, is the moral (right) thing to do. This approach requires much greater and deeper efforts than sharing information and making public service announcements. It entails a moral dialogue, in which the public is engaged, leading to a change in what people expect from one another. Among the devices that could help change the moral culture are a public statement, endorsed by community members and leaders, that expresses the community sense that donation "is what a good person does" and a community-specific web page that lists those who have made the commitment. A change in law so that a person's wishes in the matter are recognized as final and binding are also desired. This position paper deals only with cadaver organs and not living donors.


Vardenafil improves erectile function in men with erectile dysfunction irrespective of disease severity and disease classification.
Vardenafil (Levitra) is a potent and selective phosphodiesterase 5 (PDE5) inhibitor used in the management of erectile dysfunction (ED). This retrospective subgroup analysis assessed the effectiveness of vardenafil treatment in men with ED of different baseline severity and disease classification. Data from two pivotal, randomized, double-blind, placebo-controlled clinical trials enrolling men from the general ED population who received placebo or vardenafil 5 mg, 10 mg, or 20 mg during a 12-week treatment period were retrospectively analysed, stratifying by psychogenic, organic, and mixed ED disease classification as determined by the investigator. Efficacy endpoints included the International Index of Erectile Function (IIEF)-Erectile Function (EF) domain score, per-patient diary response rates to questions on penile insertion [Sexual Encounter Profile (SEP-2)] and maintenance of erection (SEP-3) and rates of positive response to the Global Assessment Question (GAQ). Data from 1,385 men who received at least one dose of study medication and had pre- and post-baseline measures of efficacy available (intent-to-treat population) are presented. At baseline 37-41% of patients had severe ED, 30-34% moderate, 22% mild-to-moderate and 6-8% mild ED. At baseline, 46-51% of patients were considered to have an organic cause for ED, 13-16% psychogenic ED, and 36-38% mixed classification of ED. For all classifications and for mild-to-moderate to severe ED, men treated with 10 or 20 mg of vardenafil showed statistically and clinically significant improvements (P < 0.001) in IIEF-EF scores, diary response rates to the SEP-2 and SEP-3 questions, and GAQ as compared with those given placebo. The greatest improvements relative to placebo were noted in patients with more severe ED. The most common treatment-emergent adverse events were headache, flushing, rhinitis, dyspepsia, and were dose-related, mostly mild to moderate in intensity and consistent with the class. Vardenafil improves EF in men with ED irrespective of investigator-determined classification and baseline ED severity.


Caspase-9 activation and caspase cleavage of tau in the Alzheimer's disease brain.
Accumulating evidence supports a role for the activation of proteolytic enzymes, caspases, in the Alzheimer's disease (AD) brain. Neurons committed to apoptosis may do so through a mitochondrial pathway employing caspase-9 or through an alternative, receptor-mediated pathway involving caspase-8. Considering the role of mitochondrial dysfunction in AD, we examined the possible activation of caspase-9 in the AD brain using an antibody that recognizes the active fragments of caspase-9, but not the full-length proform of the enzyme. In vivo immunohistochemical analysis demonstrated little caspase-9 activation in the majority of hippocampal brain sections from control brains. However, labeling of neurons as well as dystrophic neurites within plaque regions was observed in all AD hippocampal brain sections examined. In addition, active caspase-9 was colocalized with active caspase-8 and the accumulation of caspase-3-cleavage products of fodrin. The activation of caspase-9 was also observed in neurons positive for oxidative damage to DNA/RNA. A quantitative analysis indicates that as the number of neurons containing neurofibrillary tangles (NFTs) increases, the extent of caspase-9 activation decreases, supporting the idea that caspase-9 activation may precede NFT formation. In addition, a site-directed caspase-cleavage antibody was designed to the amino-terminal caspase-3 consensus cleavage site located in tau, and shown to be an effective marker for caspase-cleaved fragments of tau in vitro. Analysis with this antibody using age-matched control or AD brain sections demonstrated no staining in control brains while widespread labeling of NFTs, neuropil threads, and dystrophic neurites was observed in AD sections. Taken together, these results demonstrate the activation of caspases and cleavage of tau in the AD brain, events which may precede and lead to the formation of NFTs.


E2F-1 regulation by an unusual DNA damage-responsive DP partner subunit.
E2F activity is negatively regulated by retinoblastoma protein (pRb) through binding to the E2F-1 subunit. Within the E2F heterodimer, DP proteins are E2F partner subunits that allow proper cell cycle progression. In contrast to the other DP proteins, the newest member of the family, DP-4, downregulates E2F activity. In this study we report an unexpected role for DP-4 in regulating E2F-1 activity during the DNA damage response. Specifically, DP-4 is induced in DNA-damaged cells, upon which it binds to E2F-1 as a non-DNA-binding E2F-1/DP-4 complex. Consequently, depleting DP-4 in cells re-instates E2F-1 activity that coincides with increased levels of chromatin-bound E2F-1, E2F-1 target gene expression and associated apoptosis. Mutational analysis of DP-4 highlighted a C-terminal region, outside the DNA-binding domain, required for the negative control of E2F-1 activity. Our results define a new pathway, which acts independently of pRb and through a biochemically distinct mechanism, involved in negative regulation of E2F-1 activity.


Initial Experience With GlideAssist to Facilitate Advancement of Orbital Atherectomy Prior to Plaque Modification of Severely Calcified Coronary Artery Lesions.
We report our initial experience with GlideAssist (Cardiovascular Systems, Inc) to facilitate advancement of the orbital atherectomy crown prior to plaque modification of severely calcified coronary artery lesions. Severe coronary artery calcification increases the complexity of percutaneous coronary intervention (PCI) and is also associated with worse clinical outcomes compared with PCI of non-calcified vessels. Orbital atherectomy is an effective tool to modify calcified plaque prior to stenting. However, advancement of the orbital atherectomy crown may be technically challenging due to complex coronary anatomy. From February 2018 to February 2019, GlideAssist was used in 13 patients at the University of California, Los Angeles Medical Center. The primary endpoint was the 30-day rate of major adverse cardiac and cerebrovascular events, which was the composite of death, myocardial infarction (MI), target-vessel revascularization (TVR), and stroke. Reasons for use of GlideAssist included severe angulation/tortuosity (76.9%), ostial lesion (15.4%), and presence of previously implanted stent proximal to the calcified target lesion (7.7%). All patients who required GlideAssist had successful delivery of the crown to the calcified lesion. One patient experienced a major adverse cardiac and cerebrovascular event, which was due to periprocedural MI that was due to coronary dissection. The same patient experienced subacute stent thrombosis 13 days after the index PCI requiring TVR. No patient died or had a stroke. No other angiographic complication occurred. The GlideAssist function is a useful feature of the orbital atherectomy system to facilitate successful delivery of the crown in complex coronary anatomy.


Arterial abnormalities identified in kidneys transplanted into children during the COVID-19 pandemic.
Graft artery stenosis can have a significant short- and long-term negative impact on renal graft function. From the beginning of the COVID-19 pandemic, we noticed an unusual number of graft arterial anomalies following kidney transplant (KTx) in children. Nine children received a KTx at our center between February and July 2020, eight boys and one girl, of median age of 10 years. Seven presented Doppler features suggesting arterial stenosis, with an unusual extensive pattern. For comparison, over the previous 5-year period, persistent spectral Doppler arterial anomalies (focal anastomotic stenoses) following KTx were seen in 5% of children at our center. We retrospectively evidenced severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in five of seven children with arterial stenosis. The remaining two patients had received a graft from a deceased adolescent donor with a positive serology at D0. These data led us to suspect immune postviral graft vasculitis, triggered by SARS-CoV-2. Because the diagnosis of COVID-19 is challenging in children, we recommend pretransplant monitoring of graft recipients and their parents by monthly RT-PCR and serology. We suggest balancing the risk of postviral graft vasculitis against the risk of prolonged dialysis when considering transplantation in a child during the pandemic.


Population and Environment.
The interactions between human population dynamics and the environment have often been viewed mechanistically. This review elucidates the complexities and contextual specificities of population-environment relationships in a number of domains. It explores the ways in which demographers and other social scientists have sought to understand the relationships among a full range of population dynamics (e.g., population size, growth, density, age and sex composition, migration, urbanization, vital rates) and environmental changes. The chapter briefly reviews a number of the theories for understanding population and the environment and then proceeds to provide a state-of-the-art review of studies that have examined population dynamics and their relationship to five environmental issue areas. The review concludes by relating population-environment research to emerging work on human-environment systems.


High seroprevalence but short-lived immune response to SARS-CoV-2 infection in Paris.
Although the COVID-19 pandemic peaked in March/April 2020 in France, the prevalence of infection is barely known. Using high-throughput methods, we assessed herein the serological response against the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) of 1847 participants working in three sites of an institution in Paris conurbation. In May-July 2020, 11% (95% confidence interval [CI]: 9.7-12.6) of serums were positive for IgG against the SARS-CoV-2 N and S proteins, and 9.5% (95% CI: 8.2-11.0) were neutralizer in pseudo-typed virus assays. The prevalence of seroconversion was 11.6% (95% CI: 10.2-13.2) when considering positivity in at least one assay. In 5% of RT-qPCR positive individuals, no systemic IgGs were detected. Among immune individuals, 21% had been asymptomatic. Anosmia (loss of smell) and ageusia (loss of taste) occurred in 52% of the IgG-positive individuals and in 3% of the negative ones. In contrast, 30% of the anosmia-ageusia cases were seronegative, suggesting that the true prevalence of infection may have reached 16.6%. In sera obtained 4-8 weeks after the first sampling, anti-N and anti-S IgG titers and neutralization activity in pseudo-virus assay declined by 31%, 17%, and 53%, resulting thus in half-life of 35, 87, and 28 days, respectively. The population studied is representative of active workers in Paris. The short lifespan of the serological systemic responses suggests an underestimation of the true prevalence of infection.


Asymptomatic hyperglycemia is associated with lipid and lipoprotein changes favoring atherosclerosis.
We studied lipid and lipoprotein concentrations and their relationships to insulin level in 994 men and 1246 women ages 50 to 91 years in the upper middle-class community of Rancho Bernardo in southern California. Altogether, 593 men and 741 women had normal glucose tolerance, 240 men and 348 women, impaired glucose tolerance (IGT), 104 men and 117 women, newly diagnosed noninsulin-dependent diabetes (NIDDM), and 57 men and 40 women, previously diagnosed NIDDM. In women but not men, total cholesterol and low density lipoprotein were significantly higher in those with newly diagnosed NIDDM, compared to subjects with normal glucose tolerance. In both men and women, high density lipoprotein (HDL) cholesterol was significantly lower, and total triglyceride significantly higher, in subjects with IGT and NIDDM compared to those with normal glucose tolerance; these differences persisted after adjusting for age, body mass index, smoking, alcohol intake, and exercise level. Multiple linear-regression analyses showed that fasting insulin (but not 2-hour insulin) was significantly associated with low HDL cholesterol and high total triglycerides independently from other variables (age, body mass index, waist/hip ratio, alcohol intake, smoking, exercise, and 2-hour glucose). Overall, these results show that asymptomatic hyperglycemia (IGT, newly diagnosed NIDDM) is associated with lipid and lipoprotein changes favoring atherosclerosis and that fasting hyperinsulinemia (insulin resistance) is the most important factor associated with these lipid and lipoprotein abnormalities


Basal lamina visualization using color image processing and pattern recognition.
In histologic assessment, the absence of basal lamina is a useful feature for distinguishing invasive malignancy from benign and in situ lesions. As this feature is not possible to assess in routine H&E sections, pathologists have instead relied on histochemical and immunohistochemical stains to show components of the basal lamina such as laminin or type IV collagen. Standard image-processing software with the necessary image-processing toolbox (Matlab v5, Mathworks, Natick, MA) was used in a unique combination of color image processing and pattern recognition techniques to accentuate the collagenous stroma surrounding glands, which approximates basal lamina, in a series of benign, in situ, and invasive breast proliferations. Distinct differences in pattern were found between benign and invasive lesions, and also between in situ and malignant lesions, corresponding to that observed with type IV collagen immunostaining. Compared with immunostaining, this computer-generated method had a sensitivity of 0.96, specificity of 0.89, positive predictive value of 0.92, negative predictive value of 0.89, positive likelihood ratio of 9.1, and negative likelihood ratio of 0.042. Digital image processing serves as a less expensive and faster way of visualizing basal lamina and represents a useful adjunct to identify invasive malignancy in routinely stained sections. In addition, digital visualization of basal lamina is readily amenable to quantitative assessment, and the method provides a basis for the development of computer-based cancer diagnosis.


Exfoliation and dispersion of boron nitride nanosheets to enhance ordinary Portland cement paste.
Exfoliation and dispersion of boron nitride nanosheets (BNNSs) is the key to achieving desired reinforcing effects for ordinary Portland cement (OPC). Few studies exist, however, of the dispersion of BNNSs in a cement-induced alkaline environment and their effect on the mechanical properties of normal OPC paste. In this study, protocols were developed to prepare BNNS-reinforced OPC paste. Ultrasonication was used to exfoliate BNNSs from h-BN in water based suspensions. The addition of surfactants in the suspension was found to hinder the exfoliation of the BNNSs. The surfactants were, however, found to be essential for the dispersion of the BNNSs in pore solution. Among the three surfactants used in this study, polycarboxylate based superplasticizer was most suitable as it maintained over 40% of the BNNSs stable in the pore solution for 4 hours and increased the hydration flow peak over 20%. Atomic force microscopy results indicated that the thickness of the BNNSs was mostly under 10 nm. With the addition of 0.003 wt% BNNSs, the compressive and tensile strengths of the cement were increased by 13% and 8%, respectively. Besides the nucleation effect as indicated by hydration heat, pore structure refinement and chemical bonding were also found as the main reinforcing mechanisms of BNNSs in OPC matrix.


Americans' awareness, knowledge, and behaviors regarding fats: 2006-2007.
In recent years, epidemiologic and clinical studies, public and regulatory policy activity, and media coverage have focused on issues related to trans fats. To help increase awareness and understanding of trans fats and other fats, the American Heart Association (AHA) launched the "Face the Fats" national consumer education campaign in April 2007. The AHA commissioned a quantitative tracking survey between 2006 and 2007 to measure changes in consumer awareness, knowledge, and behaviors related fats and oils and their perceived impact on heart disease. The survey was conducted by Cogent Research. Data were collected during March 2006 and May 2007. At both time points, the survey included a representative sample of the American population age 18 to 65 years (n=1,000). The sampling plan for the survey was designed based on the 2000 and 2003 US Census. The margin of error was +/-3.10 percentage points. Awareness of trans fats increased during the 1-year study period. In 2007, 92% of respondents were aware of trans fats, an increase from 84% in 2006 (P<0.05). The 2007 level was similar to the awareness of saturated fats (93%). Perceptions that certain fats and oils heighten the risk of heart disease increased for trans fats (73% in 2007 vs 63% in 2006; P<0.05), saturated fats (77% in 2007 vs 73% in 2006; P<0.05), and partially hydrogenated oils (56% in 2007 vs 49% in 2006; P<0.05). Knowledge about food sources of different fats remained low. On an unaided basis, 21% could name three food sources of trans fats in 2007, up from 17% in 2006 (P<0.05). Knowledge of food sources of saturated fat remained unchanged at 30% in 2007. Significantly more respondents in 2007 reported behavioral changes related to trans fat information, such as buying food products because they show "zero trans fat" on labels or packages (37% in 2007 vs. 32% in 2006; P<0.05). Between 2006 and 2007, consumer awareness about trans fats increased and attained awareness levels similar to saturated fats. The increased awareness is associated with improved self-reported behaviors in grocery shopping. Nonetheless, overall knowledge, especially regarding food sources of saturated and trans fats, remains relatively low, underscoring the need for heightened consumer education activities. The positive change in consumer awareness about trans fats is likely attributable to the wide range of messages available to them, including the AHA "Face the Fats" national consumer education campaign.


Community health workers in Canada: innovative approaches to health promotion outreach and community development among immigrant and refugee populations.
This article provides results from an empirical case study that showcases a community health worker practice targeting immigrants and refugees in Canada. The study focuses on the Multicultural Health Brokers practice, which offers an innovative approach to health promotion outreach and community development addressing broad social determinants of health. This article offers new evidence of both the role of community health worker interventions in Canada and community health workers as an invisible health and human services workforce. It also discusses the Multicultural Health Brokers contribution both to the "new public health" vision in Canada and to a practice that fosters feminist urban citizenship.


Lifestyle intervention leading to moderate weight loss normalizes postprandial triacylglycerolemia despite persisting obesity.
Obesity is associated with impaired postprandial triacylglycerolemia, an independent risk factor for cardiovascular disease. Given that obesity is hard to treat, efforts should focus on treating its comorbidities. We aimed to investigate whether moderate weight loss normalizes postprandial triacylglycerol (TAG) concentrations, in the absence of the acute effects of negative energy balance. For this purpose, postprandial lipemia was investigated in eight obese but otherwise healthy, sedentary men (age: 41.3 ± 4.1 years, BMI: 36.5 ± 1.6 kg·m(-2)), once before and again after a 10% weight loss followed by ≥4 weeks of weight maintenance, and was compared with that of eight age-matched healthy lean men (BMI: 24.7 ± 0.6 kg·m(-2)). Dietary intervention consisted of reduced carbohydrate and saturated fat intake and increased monounsaturated fat intake. Obese volunteers were advised to increase physical activity using pedometers to record daily activity. Postprandial triacylglycerolemia after weight loss was reduced by 27-46% (P < 0.05), and became similar to that of lean men despite persisting obesity (BMI after weight loss: 32.9 ± 1.5 kg·m(-2)). Reduction in postprandial TAG responses was inversely correlated with the decrease in postprandial insulin sensitivity index (ISI) after weight loss (r = -0.714, P = 0.047). We conclude that moderate weight loss induced by a low-carbohydrate and saturated fat diet and a slight increase in daily physical activity normalizes postprandial triacylglycerolemia in obese men, independently of acute diet-induced negative energy balance, and possibly through enhancement of insulin action.


Shame, Guilt, and Suicide Ideation among Bondage and Discipline, Dominance and Submission, and Sadomasochism Practitioners: Examining the Role of the Interpersonal Theory of Suicide.
To date, no study has examined rates of suicide ideation or theory-based risk factors for suicide ideation among bondage and discipline, dominance and submission, and sadomasochism (BDSM) practitioners. Participants were 321 adults that endorsed BDSM involvement. Thirty-seven percent of the sample indicated a nonzero level of suicide ideation. Thwarted belongingness and perceived burdensomeness (PB) were positively associated with suicide ideation and their interactive effect predicted additional variance in suicide ideation after adjusting for depressive symptoms. Overall, shame and guilt were positively associated with suicide ideation and these relations were mediated by thwarted belongingness and PB in parallel adjusting for depressive symptoms; however, there were some differences between demographic subgroups. Among BDSM practitioners, stigma-related internalized feelings (i.e., shame and guilt) may be associated with increased thwarted belongingness and PB, which are associated with suicide ideation.


Variation of fluorescence spectroscopy during the menstrual cycle.
Cervical autofluorescence has been demonstrated to have potential for real-time diagnosis. Inter-patient and intra-patient variations in fluorescence intensity have been measured. Inter-patient measurements may vary by a factor of ten, while intra-patient measurements may vary by a factor of two. Age and menopausal status have been demonstrated to account for some of the variations, while race and smoking have not. In order to explore in detail the role of the menstrual cycle in intra-patient variation, a study was designed to measure fluorescence excitation emission matrices (EEMs) in patients daily throughout one cycle. Ten patients with a history of normal menstrual cycles and normal Papanicolaou smears underwent daily measurements of fluorescence EEMs from three colposcopically normal sites throughout one menstrual cycle. Changes in signals from porphyrin, NADH, and FAD fluorescence and blood absorption were noted when the data was viewed in a graphical format. Visually interpreted features of the EEMs in this graphical format did not appear to correlate with the day of the menstrual cycle with the exception that blood absorption features were more prominent during the menstrual phase (during which bleeding occurs), suggesting that measurements during the menstrual phase should be avoided. Variations in cycle date likely do not account for inter- or intra-patient variations.


O6-methylguanine induces altered proteins at the level of transcription in human cells.
O(6)-Methylguanine (O(6)-meG), which is produced in DNA following exposure to methylating agents, instructs human RNA polymerase II to mis-insert bases opposite the lesion during transcription. In this study, we examined the effect of O(6)-meG on transcription in human cells and investigated the subsequent effects on protein function following translation of the resulting mRNA. In HEK293 cells, O(6)-meG induced incorporation of uridine or cytidine in nascent RNA opposite the adduct. In cells containing active O(6)-alkylguanine-DNA alkyltransferase (AGT), which repairs O(6)-meG, 3% misincorporation of uridine was observed opposite the lesion. In cells where AGT function was compromised by addition of the AGT inhibitor O(6)-benzylguanine, ∼ 58% of the transcripts contained a uridine misincorporation opposite the lesion. Furthermore, the altered mRNA induced changes to protein function as demonstrated through recovery of functional red fluorescent protein (RFP) from DNA coding for a non-fluorescent variant of RFP. These data show that O(6)-meG is highly mutagenic at the level of transcription in human cells, leading to an altered protein load, especially when AGT is inhibited.


The golden hour: scientific fact or medical "urban legend"?
The term "golden hour" is commonly used to characterize the urgent need for the care of trauma patients. This term implies that morbidity and mortality are affected if care is not instituted within the first hour after injury. This concept justifies much of our current trauma system. However, definitive references are generally not provided when this concept is discussed. It remains unclear whether objective data exist. This article discusses a detailed literature and historical record search for support of the "golden hour" concept. None is identified.


Association of the use of statins with disease activity and functional status in Puerto Ricans with rheumatoid arthritis.
Statins, which appear to have anti-inflammatory and immunomodulatory effects, may benefit patients with rheumatoid arthritis (RA). Our study sought to determine the association of statins use with disease activity and functional status in a group of patients with RA. A cross-sectional study was performed in 209 Puerto Ricans with RA (per the 1987 classification criteria of the American College of Rheumatology). Demographic features, lifestyle-related behaviors, disease activity (per Disease Activity Score 28), comorbid conditions, functional status (per Health Assessment Questionnaire), pharmacologic therapy, and patients' and physicians' global assessments using visual analogue scales, were determined. Data were examined using univariate, bivariate, and multiple logistic regression analyses. The mean (standard deviation [SD]) age of the study population at study visit was 56.8 (13.5) years (range: 24-86 years); 175 patients (83.7%) were women. The mean (SD) disease duration was 10.4 (9.5) years (range: 0.0-44.0 years). Thirty-two (15.3%) patients were using statins at study visit, and 36 (17.2%) had used statins in the past. In the multivariable analysis, the current use of statins was associated with higher functional status (odds ratio 0.42, 95% confidence interval 0.22-0.80) than was nonuse, after adjusting for age, disease duration, arterial hypertension, coronary artery disease, and dyslipidemia. No association between either current or past use of statins and disease activity was found. In this group of RA patients, the current use of statins was associated with a higher functional status; conversely, no association was found between statins use and disease activity. However, larger and longitudinal studies are required to confirm these findings.


Escherichia coli O157:H7: recent advances in research on occurrence, transmission, and control in cattle and the production environment.
Escherichia coli O157:H7 is a zoonotic pathogen that is an important cause of human foodborne and waterborne disease, with a spectrum of illnesses ranging from asymptomatic carriage and diarrhea to the sometimes fatal hemolytic uremic syndrome. Outbreaks of E. coli O157:H7 disease are often associated with undercooked beef, but there are other sources of transmission, including water, produce, and animal contact, which can often be linked directly or indirectly to cattle. Thus, preharvest control of this pathogen in cattle production should have a large impact on reducing the risk of human foodborne illness. In this review, we will summarize preharvest research on E. coli O157:H7 in cattle and the production environment, focusing on factors that may influence the transmission, prevalence, and levels of this pathogen, such as season, diet, high-level shedders, and animal stress. In addition, we will discuss recent research on the reduction of this pathogen in cattle production, including vaccination, probiotics, bacteriophage, and manure treatments.


Optimization of multi-grating volume holographic spectrum splitters for photovoltaic applications.
Recent research has shown that using multiple diverse-bandgap photovoltaic (PV) cells in conjunction with a spectrum splitting optical system can significantly improve PV power generation efficiency. Although volume Bragg gratings (VBGs) can serve as effective spectrum splitters, the inherent dispersion of a VBG can be detrimental given a broad-spectrum input. The performance of a single holographic spectrum splitter element can be improved by utilizing multiple single volume gratings, each operating in a slightly different spectral band. However, care must be taken to avoid inter-grating coupling effects that limit the ultimate performance. This work explores broadband two-grating holographic optical elements (HOEs) in multiplexed (single element) and sandwiched-grating arrangements. Particle swarm optimization is used to tailor these systems to the solar spectrum, taking into account both efficiency and dispersion. Both multiplexed and sandwiched two-grating systems exhibit performance improvements over single-grating solutions, especially when reduced dispersion is required. Under a ±2° constraint on output angular spread from wavelength dispersion, sandwiched-, multiplexed-, and single-grating systems exhibit power conversion efficiencies of 82.1%, 80.9%, and 77.5%, respectively, compared to an ideal bandpass spectrum splitter. Dispersion performance can be further improved by employing more than two VBGs in the spectrum splitter, but efficiency is compromised by additional cross-coupling effects. Multiplexed-grating systems are especially susceptible to these effects, but have the advantage of utilizing only a single HOE.


The robustness of proofreading to crowding-induced pseudo-processivity in the MAPK pathway.
Double phosphorylation of protein kinases is a common feature of signaling cascades. This motif may reduce cross-talk between signaling pathways because the second phosphorylation site allows for proofreading, especially when phosphorylation is distributive rather than processive. Recent studies suggest that phosphorylation can be pseudo-processive in the crowded cellular environment, since rebinding after the first phosphorylation is enhanced by slow diffusion. Here, we use a simple model with unsaturated reactants to show that specificity for one substrate over another drops as rebinding increases and pseudo-processive behavior becomes possible. However, this loss of specificity with increased rebinding is typically also observed if two distinct enzyme species are required for phosphorylation, i.e., when the system is necessarily distributive. Thus the loss of specificity is due to an intrinsic reduction in selectivity with increased rebinding, which benefits inefficient reactions, rather than pseudo-processivity itself. We also show that proofreading can remain effective when the intended signaling pathway exhibits high levels of rebinding-induced pseudo-processivity, unlike other proposed advantages of the dual phosphorylation motif.


Infectiousness of the human population to Anopheles arabiensis by direct skin feeding in an area hypoendemic for malaria in Senegal.
Direct skin feeding experiments are sensitive assays to determine human infectiousness to mosquitoes but are rarely used in malaria epidemiological surveys. We determined the infectiousness of inhabitants of a malaria hypoendemic area in Senegal. Gametocyte prevalence by microscopy was 13.5% (26 of 192). Of all individuals who were gametocyte positive, 44.4% (11 of 25) infected ≥ 1 Anopheles arabiensis mosquito and 10.8% (54 of 500) of mosquitoes became infected. Of all individuals who were gametocyte negative by microscopy, 4.3% (7 of 162) infected ≥ 1 mosquito and 0.4% (12 of 3240) of mosquitoes became infected. The 18.2% (12 of 66) of all mosquito infections was a result of submicroscopic gametocyte carriage and two individuals without asexual parasites or gametocytes by microscopy were infectious to mosquitoes. When infectivity and local demography was taken into account, children 5-14 years of age contributed 50.8% of the human infectious reservoir for malaria. Adults and submicroscopic gametocyte carriers may contribute considerably to onward malaria transmission in our setting.


Comparison of T Cell Activities Mediated by Human TCRs and CARs That Use the Same Recognition Domains.
Adoptive T cell therapies have achieved significant clinical responses, especially in hematopoietic cancers. Two types of receptor systems have been used to redirect the activity of T cells, normal heterodimeric TCRs or synthetic chimeric Ag receptors (CARs). TCRs recognize peptide-HLA complexes whereas CARs typically use an Ab-derived single-chain fragments variable that recognizes cancer-associated cell-surface Ags. Although both receptors mediate diverse effector functions, a quantitative comparison of the sensitivity and signaling capacity of TCRs and CARs has been limited due to their differences in affinities and ligands. In this study we describe their direct comparison by using TCRs that could be formatted either as conventional αβ heterodimers, or as single-chain fragments variable constructs linked to CD3ζ and CD28 signaling domains or to CD3ζ alone. Two high-affinity TCRs (KD values of ∼50 and 250 nM) against MART1/HLA-A2 or WT1/HLA-A2 were used, allowing MART1 or WT1 peptide titrations to easily assess the impact of Ag density. Although CARs were expressed at higher surface levels than TCRs, they were 10-100-fold less sensitive, even in the absence of the CD8 coreceptor. Mathematical modeling demonstrated that lower CAR sensitivity could be attributed to less efficient signaling kinetics. Furthermore, reduced cytokine secretion observed at high Ag density for both TCRs and CARs suggested a role for negative regulators in both systems. Interestingly, at high Ag density, CARs also mediated greater maximal release of some cytokines, such as IL-2 and IL-6. These results have implications for the next-generation design of receptors used in adoptive T cell therapies.


Phosphatidylglycerol::prolipoprotein diacylglyceryl transferase (Lgt) of Escherichia coli has seven transmembrane segments, and its essential residues are embedded in the membrane.
Lgt of Escherichia coli catalyzes the transfer of an sn-1,2-diacylglyceryl group from phosphatidylglycerol to prolipoproteins. The enzyme is essential for growth, as demonstrated here by the analysis of an lgt depletion strain. Cell fractionation demonstrated that Lgt is an inner membrane protein. Its membrane topology was determined by fusing Lgt to β-galactosidase and alkaline phosphatase and by substituted cysteine accessibility method (SCAM) studies. The data show that Lgt is embedded in the membrane by seven transmembrane segments, that its N terminus faces the periplasm, and that its C terminus faces the cytoplasm. Highly conserved amino acids in Lgt of both Gram-negative and Gram-positive bacteria were identified. Lgt enzymes are characterized by a so-called Lgt signature motif in which four residues are invariant. Ten conserved residues were replaced with alanine, and the activity of these Lgt variants was analyzed by their ability to complement the lgt depletion strain. Residues Y26, N146, and G154 are absolutely required for Lgt function, and R143, E151, R239, and E243 are important. The results demonstrate that the majority of the essential residues of Lgt are located in the membrane and that the Lgt signature motif faces the periplasm.


Recent Secondary Contacts, Linked Selection, and Variable Recombination Rates Shape Genomic Diversity in the Model Species Anolis carolinensis.
Gaining a better understanding on how selection and neutral processes affect genomic diversity is essential to gain better insights into the mechanisms driving adaptation and speciation. However, the evolutionary processes affecting variation at a genomic scale have not been investigated in most vertebrate lineages. Here, we present the first population genomics survey using whole genome resequencing in the green anole (Anolis carolinensis). Anoles have been intensively studied to understand mechanisms underlying adaptation and speciation. The green anole in particular is an important model to study genome evolution. We quantified how demography, recombination, and selection have led to the current genetic diversity of the green anole by using whole-genome resequencing of five genetic clusters covering the entire species range. The differentiation of green anole's populations is consistent with a northward expansion from South Florida followed by genetic isolation and subsequent gene flow among adjacent genetic clusters. Dispersal out-of-Florida was accompanied by a drastic population bottleneck followed by a rapid population expansion. This event was accompanied by male-biased dispersal and/or selective sweeps on the X chromosome. We show that the interaction between linked selection and recombination is the main contributor to the genomic landscape of differentiation in the anole genome.


Genetic association study of childhood aggression across raters, instruments, and age.
Childhood aggressive behavior (AGG) has a substantial heritability of around 50%. Here we present a genome-wide association meta-analysis (GWAMA) of childhood AGG, in which all phenotype measures across childhood ages from multiple assessors were included. We analyzed phenotype assessments for a total of 328 935 observations from 87 485 children aged between 1.5 and 18 years, while accounting for sample overlap. We also meta-analyzed within subsets of the data, i.e., within rater, instrument and age. SNP-heritability for the overall meta-analysis (AGGoverall) was 3.31% (SE = 0.0038). We found no genome-wide significant SNPs for AGGoverall. The gene-based analysis returned three significant genes: ST3GAL3 (P = 1.6E-06), PCDH7 (P = 2.0E-06), and IPO13 (P = 2.5E-06). All three genes have previously been associated with educational traits. Polygenic scores based on our GWAMA significantly predicted aggression in a holdout sample of children (variance explained = 0.44%) and in retrospectively assessed childhood aggression (variance explained = 0.20%). Genetic correlations (rg) among rater-specific assessment of AGG ranged from rg = 0.46 between self- and teacher-assessment to rg = 0.81 between mother- and teacher-assessment. We obtained moderate-to-strong rgs with selected phenotypes from multiple domains, but hardly with any of the classical biomarkers thought to be associated with AGG. Significant genetic correlations were observed with most psychiatric and psychological traits (range [Formula: see text]: 0.19-1.00), except for obsessive-compulsive disorder. Aggression had a negative genetic correlation (rg = ~-0.5) with cognitive traits and age at first birth. Aggression was strongly genetically correlated with smoking phenotypes (range [Formula: see text]: 0.46-0.60). The genetic correlations between aggression and psychiatric disorders were weaker for teacher-reported AGG than for mother- and self-reported AGG. The current GWAMA of childhood aggression provides a powerful tool to interrogate the rater-specific genetic etiology of AGG.


Molecular motors: how to make models that can be used to convey the concept of molecular ratchets and thermal capture.
A wide variety of cellular processes use molecular motors, including processive motors that move along some form of track (e.g., myosin with actin, kinesin or dynein with tubulin) and polymerases that move along a template (e.g., DNA and RNA polymerases, ribosomes). In trying to understand how these molecular motors actually move, many apply their understanding of how man-made motors work: the latter use some form of energy to exert a force or torque on its load. However, quite a different mechanism has been proposed to possibly account for the movement of molecular motors. Rather than hydrolyzing ATP to push or pull their load, they might use their own thermal vibrational energy as well as that of their load and their environment to move the load, capturing those movements that occur along a desired vector or axis and resisting others; ATP hydrolysis is required to make backward movements impossible. This intriguing thermal capture or Brownian ratchet model is relatively more difficult to convey to students. In this report, we describe several teaching aids that are very easily constructed using widely available household materials to convey the concept of a molecular ratchet.


Obesity, albuminuria and hypertension among Hong Kong Chinese with non-insulin-dependent diabetes mellitus (NIDDM).
A total of 412 Hong Kong Chinese diabetic patients were studied on at least two occasions 8-16 weeks apart. Although 28% were insulin-treated, only 3.6% had insulin-dependent diabetes (IDDM). In the remaining 397 patients with non-insulin-dependent diabetes (NIDDM), the mean (s.d.) body mass index (BMI) was 24.4 +/- 3.2 kg/m2 in females and 24.2 +/- 3.2 kg/m2 in males. Obesity was present in 17% of males (BMI > 27 kg/m2) and 40% of females (BMI > 25 kg/m2). Established hypertension was present in 49%. Abnormal albuminuria, defined as a mean urinary albumin/creatinine (UA/Cr) ratio greater than 5.4 mg/mmol based on two random spot urine samples, was present in 47%. On stepwise multiple regression analysis, UA/Cr ratio (R2 = 0.34, F = 65.4, P < 0.001) showed significant associations with systolic blood pressure (standardized regression coefficient beta = 0.40, P < 0.001), plasma creatinine concentration (beta = 0.27, P < 0.001) and glycosylated haemoglobin (beta = 0.20, P < 0.001). While the prevalence of hypertension increased with increasing severity of proteinuria, 40% of normoalbuminuric patients had hypertension. Among patients diagnosed before the age of 35 (n = 67), 52% were insulin-treated although only 10% were insulin-dependent. Among these NIDDM patients of young onset (n = 59), obesity was present in 25% of males and 56% of females. Overall, 18% of these patients had a blood pressure greater than 140/90 mmHg and 27% had abnormal albuminuria. In Hong Kong Chinese, diabetes mellitus is predominantly non-insulin-dependent even in the young. Obesity is more prevalent among females. Abnormal albuminuria is relatively common and is closely associated with hypertension and glycaemic control. In the light of increasing prevalence of diabetes among overseas Chinese, our findings may have important implications in the management of Chinese diabetic patients.


Physicochemical properties of air discharge-generated manganese oxide nanoparticles: Comparison to welding fumes.
Exposures to high doses of manganese (Mn) via inhalation, dermal contact or direct consumption can cause adverse health effects. Welding fumes are a major source of manganese containing nanoparticles in occupational settings. Understanding the physicochemical properties of manganese-containing nanoparticles can be a first step in understanding their toxic potential following exposure. In particular, here we compare the size, morphology and Mn oxidation states of Mn oxide nanoparticles generated in the laboratory by arc discharge to those from welding collected in heavy vehicle manufacturing. Fresh nanoparticles collected at the exit of the spark discharge generation chamber consisted of individual or small aggregates of primary particles. These nanoparticles were allowed to age in a chamber to form chain-like aggregates of primary particles with morphologies very similar to welding fumes. The primary particles were a mixture of hausmannite (Mn3O4), bixbyite (Mn2O3) and manganosite (MnO) phases, whereas aged samples revealed a more amorphous structure. Both Mn2+ and Mn3+, as in double valence stoichiometry present in Mn3O4, and Mn3+, as in Mn2O3 and MnOOH, were detected by X-ray photoelectron spectroscopy on the surface of the nanoparticles in the laboratory nanoparticles and welding fumes. Dissolution studies conducted for these two Mn samples (aged and fresh fume) reveal different release kinetics of Mn ions in artificial lysosomal fluid (pH 4.5) and very limited dissolution in Gamble's solution (pH 7.4). Taken together, these data suggest several important considerations for understanding the health effects of welding fumes. First, the method of particle generation affects the crystallinity and phase of the oxide. Second, welding fumes consist of multiple oxidation states whether they are amorphous or crystalline or occur as isolated nanoparticles or agglomerates. Third, although the dissolution behavior depends on conditions used for nanoparticle generation, the dissolution of Mn oxide nanoparticles in the lysosome may promote Mn ions translocation into various organs causing toxic effects.


Conceptualizing alternate regimes in a large floodplain-river ecosystem: Water clarity, invasive fish, and floodplain vegetation.
Regime shifts - persistent changes in the structure and function of an ecosystem - are well-documented for some ecosystems and have informed research and management of these ecosystems. In floodplain-river ecosystems, there is growing interest from restoration practitioners in ecological resilience, yet regime shifts remain poorly understood in these ecosystems. To understand how regime shifts may apply to floodplain-river ecosystems, we synthesize our understanding of ecosystem dynamics using an alternate regimes conceptual framework. We present three plausible sets of alternate regimes relevant to natural resource management interests within the Upper Mississippi River and Illinois River. These alternate regimes include: 1) a clear water and abundant vegetation regime vs. a turbid water and sparse vegetation regime in lentic, off-channel areas, 2) a diverse native fish community regime vs. an invasive-dominated fish community regime, and 3) a regime characterized by a diverse and dynamic mosaic of floodplain vegetation types vs. one characterized as a persistent invasive wet meadow monoculture. For each set of potential alternate regimes, we review available literature to synthesize known or hypothesized feedback mechanisms that reinforce regimes, controlling variables that drive regime transitions, and current restoration pathways. Our conceptual models provide preliminary support for the existence of alternate regimes in floodplain-river ecosystems. Quantitatively testing hypotheses contained within the conceptual model are important next steps in evaluating the model. Ultimately, the synthesis and evaluation of alternate regimes can inform the utility of resilience concepts in restoration and management on the Upper Mississippi River and Illinois River and improve our understanding of ecosystem dynamics in other large, heavily managed floodplain-river ecosystems.


Mu opioid receptor activation inhibits GABAergic inputs to basolateral amygdala neurons through Kv1.1/1.2 channels.
The basolateral amygdala (BLA) is the major amygdaloid nucleus distributed with mu opioid receptors. The afferent input from the BLA to the central nucleus of the amygdala (CeA) is considered important for opioid analgesia. However, little is known about the effect of mu opioids on synaptic transmission in the BLA. In this study, we examined the effect of mu opioid receptor stimulation on the inhibitory and excitatory synaptic inputs to CeA-projecting BLA neurons. BLA neurons were retrogradely labeled with a fluorescent tracer injected into the CeA of rats. Whole cell voltage-clamp recordings were performed on labeled BLA neurons in brain slices. The specific mu opioid receptor agonist, (D-Ala2,N-Me-Phe4,Gly5-ol)-enkephalin (DAMGO, 1 microM), significantly reduced the frequency of miniature inhibitory postsynaptic currents (mIPSCs) in 77% of cells tested. DAMGO also significantly decreased the peak amplitude of evoked IPSCs in 75% of cells examined. However, DAMGO did not significantly alter the frequency of mEPSCs or the peak amplitude of evoked EPSCs in 90% and 75% of labeled cells, respectively. Bath application of the Kv channel blockers, 4-AP (Kv1.1, 1.2, 1.3, 1.5, 1.6, 3.1, 3.2), alpha-dendrotoxin (Kv1.1, 1.2, 1.6), dendrotoxin-K (Kv1.1), or tityustoxin-Kalpha (Kv1.2) each blocked the inhibitory effect of DAMGO on mIPSCs. Double immunofluorescence labeling showed that some of the immunoreactivities of Kv1.1 and Kv1.2 were colocalized with synaptophysin in the BLA. This study provides new information that activation of presynaptic mu opioid receptors primarily attenuates GABAergic synaptic inputs to CeA-projecting neurons in the BLA through a signaling mechanism involving Kv1.1 and Kv1.2 channels.


Living in a fungal world: impact of fungi on soil bacterial niche development.
The colonization of land by plants appears to have coincided with the appearance of mycorrhiza-like fungi. Over evolutionary time, fungi have maintained their prominent role in the formation of mycorrhizal associations. In addition, however, they have been able to occupy other terrestrial niches of which the decomposition of recalcitrant organic matter is perhaps the most remarkable. This implies that, in contrast to that of aquatic organic matter decomposition, bacteria have not been able to monopolize decomposition processes in terrestrial ecosystems. The emergence of fungi in terrestrial ecosystems must have had a strong impact on the evolution of terrestrial bacteria. On the one hand, potential decomposition niches, e.g. lignin degradation, have been lost for bacteria, whereas on the other hand the presence of fungi has itself created new bacterial niches. Confrontation between bacteria and fungi is ongoing, and from studying contemporary interactions, we can learn about the impact that fungi presently have, and have had in the past, on the ecology and evolution of terrestrial bacteria. In the first part of this review, the focus is on niche differentiation between soil bacteria and fungi involved in the decomposition of plant-derived organic matter. Bacteria and fungi are seen to compete for simple plant-derived substrates and have developed antagonistic strategies. For more recalcitrant organic substrates, e.g. cellulose and lignin, both competitive and mutualistic strategies appear to have evolved. In the second part of the review, bacterial niches with respect to the utilization of fungal-derived substrates are considered. Here, several lines of development can be recognized, ranging from mutualistic exudate-consuming bacteria that are associated with fungal surfaces to endosymbiotic and mycophagous bacteria. In some cases, there are indications of fungal specific selection in fungus-associated bacteria, and possible mechanisms for such selection are discussed.


Exercise-induced alterations in intramyocellular lipids and insulin resistance: the athlete's paradox revisited.
We previously reported an "athlete's paradox" in which endurance-trained athletes, who possess a high oxidative capacity and enhanced insulin sensitivity, also have higher intramyocellular lipid (IMCL) content. The purpose of this study was to determine whether moderate exercise training would increase IMCL, oxidative capacity of muscle, and insulin sensitivity in previously sedentary overweight to obese, insulin-resistant, older subjects. Twenty-five older (66.4 +/- 0.8 yr) obese (BMI = 30.3 +/- 0.7 kg/m2) men (n = 9) and women (n = 16) completed a 16-wk moderate but progressive exercise training program. Body weight and fat mass modestly but significantly (P < 0.01) decreased. Insulin sensitivity, measured using the euglycemic hyperinsulinemic clamp, was increased (21%, P = 0.02), with modest improvements (7%, P = 0.04) in aerobic fitness (Vo2peak). Histochemical analyses of IMCL (Oil Red O staining), oxidative capacity [succinate dehydrogenase activity (SDH)], glycogen content, capillary density, and fiber type were performed on skeletal muscle biopsies. Exercise training increased IMCL by 21%. In contrast, diacylglycerol and ceramide, measured by mass spectroscopy, were decreased (n = 13; -29% and -24%, respectively, P < 0.05) with exercise training. SDH (19%), glycogen content (15%), capillary density (7%), and the percentage of type I slow oxidative fibers (from 50.8 to 55.7%), all P < or = 0.05, were increased after exercise. In summary, these results extend the athlete's paradox by demonstrating that chronic exercise in overweight to obese older adults improves insulin sensitivity in conjunction with favorable alterations in lipid partitioning and an enhanced oxidative capacity within muscle. Therefore, several key deleterious effects of aging and/or obesity on the metabolic profile of skeletal muscle can be reversed with only moderate increases in physical activity.


The drug transporter P-glycoprotein limits oral absorption and brain entry of HIV-1 protease inhibitors.
Currently available HIV-1 protease inhibitors are potent agents in the therapy of HIV-1 infection. However, limited oral absorption and variable tissue distribution, both of which are largely unexplained, complicate their use. We tested the hypothesis that P-glycoprotein is an important transporter for these agents. We studied the vectorial transport characteristics of indinavir, nelfinavir, and saquinavir in vitro using the model P-glycoprotein expressing cell lines L-MDR1 and Caco-2 cells, and in vivo after intravenous and oral administration of these agents to mice with a disrupted mdr1a gene. All three compounds were found to be transported by P-glycoprotein in vitro. After oral administration, plasma concentrations were elevated 2-5-fold in mdr1a (-/-) mice and with intravenous administration, brain concentrations were elevated 7-36-fold. These data demonstrate that P-glycoprotein limits the oral bioavailability and penetration of these agents into the brain. This raises the possibility that higher HIV-1 protease inhibitor concentrations may be obtained by targeted pharmacologic inhibition of P-glycoprotein transport activity.


Selection of methods for assessing sediment toxicity in California bays and estuaries.
Toxicity tests are often used in sediment assessment programs. However, the choice of methods has been largely limited to acute tests. Where sublethal methods have been used, there has been little consistency among programs in the types of the sublethal tests used. The goal of this study was to develop a method for choosing a suite of acute and sublethal tests for use in a California statewide assessment program, and to develop a set of method-specific thresholds for classifying the degree of toxicity within a multiple line of evidence framework consisting of sediment chemistry, benthic community structure, and sediment toxicity. A group of candidate methods was evaluated using feasibility and performance criteria. Toxicity thresholds were calculated based on test variability and sensitivity. As a result of the evaluation, 3 acute toxicity methods using amphipods (Eohaustorius estuarius, Rhepoxynius abronius, and Leptocheirus plumulosus), and 2 sublethal methods using a polychaete and mussel embryos (Neanthes arenaceodentata growth and Mytilus galloprovincialis embryo development at the sediment-water interface) were selected for recommendation. Thresholds for toxicity categories corresponding to Nontoxic, Low Toxicity, Moderate Toxicity, and High Toxicity were developed for each test method. Although these toxicity categories and thresholds provide a consistent framework for the interpretation of test results among different methods, additional research is needed to determine their effectiveness for predicting impacts to benthic communities.