Do mismatches between pre- and post-natal environments influence adult physiological functioning?
Mismatches between pre- and post-natal environments have implications for disease in adulthood. However, less is known about how this mismatch can affect physiological systems more generally, especially at younger ages. We hypothesised that mismatches between pre- and post-natal environments, as measured by the measures of birthweight and adult leg length, would be associated with poorer biomarker levels across five key physiological systems in young adults. Data were collected from 923, 36 year-old respondents from the West of Scotland Twenty-07 Study. The biomarkers were: systolic blood pressure (sBP); forced expiratory volume (FEV1); glycated haemoglobin (HbA1c); glomerular filtration rate (eGFR); and gamma-glutamyltransferase (GGT). These biomarkers were regressed against pre-natal conditions (birthweight), post-natal conditions (leg length) and the interaction between pre- and post-natal measures. Sex, childhood socioeconomic position and adult lifestyle characteristics were adjusted for as potential effect modifiers and confounders, respectively. There were no associations between birthweight and leg length and sBP, FEV1, HbA1c, or GGT. Higher birthweight and longer leg length were associated with better kidney function (eGFR). However, there was no evidence for mismatches between birthweight and leg length to be associated with worse sBP, FEV1, HbA1c, eGFR or GGT levels (P>0.05). Our hypothesis that early signs of physiological damage would be present in young adults given mismatches in childhood environments, as measured by growth markers, was not proven. This lack of association could be because age 36 is too young to identify significant trends for future health, or the associations simply not being present.


First demonstration of plasmonic GaN quantum cascade detectors with enhanced efficiency at normal incidence.
We have designed, fabricated and measured the first plasmon-assisted normal incidence GaN/AlN quantum cascade detector (QCD) making use of the surface plasmon resonance of a two-dimensional nanohole Au array integrated on top of the detector absorption region. The spectral response of the detector at room temperature is peaked at the plasmon resonance of 1.82 μm. We show that the presence of the nanohole array induces an absolute enhancement of the responsivity by a factor of ~30 over that of the bare device at normal incidence and by a factor of 3 with respect to illumination by the 45° polished side facet. We show that this significant improvement arises from two phenomena, namely, the polarization rotation of the impinging light from tangential to normal induced by the plasmonic structure and from the enhancement of the absorption cross-section per quantum well due to the near-field optical intensity of the plasmonic wave.


Effects of caffeine on alcohol consumption and nicotine self-administration in rats.
Caffeine, alcohol, and nicotine are 3 of the most widespread self-administered psychoactive substances, which are known to be extensively co-administered. However, little is known about the degree to which they may mutually potentiate each other's consumption. In the current set of studies, we examined in rats the effect of caffeine administration on alcohol drinking and intravenous (i.v.) self-administration of nicotine. In male alcohol-preferring (P) rats, caffeine (5, 10, and 20 mg/kg) or the saline vehicle was administered acutely either by subcutaneous (S.C.) injection or orally (PO) by gavage. In a chronic study, the effect of PO caffeine (5 and 20 mg/kg) on alcohol intake over a 10-day period was tested. In another experiment, the effect of acute PO administration of caffeine (20 mg/kg) or saline on saccharin intake (0.2% solution) was determined in P rats. Effects of 20 mg/kg caffeine on motor activity were also determined in P rats. Finally, the effects of acute PO caffeine administration on nicotine self-administration in Sprague-Dawley rats were also determined. Both routes of administration of caffeine, S.C. and PO, caused a significant dose-related decrease in alcohol intake and preference during free access to alcohol and after 4-day deprivation of alcohol. However, the low dose of 5 mg/kg caffeine increased alcohol intake. Acute PO caffeine also reduced saccharin intake. Acute systemic administration of 20 mg/kg caffeine did not exert a significant effect on motor activity. In Sprague-Dawley rats trained to self-administer i.v. nicotine, acute PO administration of caffeine significantly increased self-administration of nicotine in a dose-related manner. These results suggest that adenosine receptor systems may play a role in both alcohol and nicotine intake and deserve further study regarding these addictions.


Novel Biochemical Markers of Glycemia to Predict Pregnancy Outcomes in Women With Type 1 Diabetes.
The optimal method of monitoring glycemia in pregnant women with type 1 diabetes remains controversial. This study aimed to assess the predictive performance of HbA1c, continuous glucose monitoring (CGM) metrics, and alternative biochemical markers of glycemia to predict obstetric and neonatal outcomes. One hundred fifty-seven women from the Continuous Glucose Monitoring in Women With Type 1 Diabetes in Pregnancy Trial (CONCEPTT) were included in this prespecified secondary analysis. HbA1c, CGM data, and alternative biochemical markers (glycated CD59, 1,5-anhydroglucitol, fructosamine, glycated albumin) were compared at ∼12, 24, and 34 weeks' gestation using logistic regression and receiver operating characteristic (ROC) curves to predict pregnancy complications (preeclampsia, preterm delivery, large for gestational age, neonatal hypoglycemia, admission to neonatal intensive care unit). HbA1c, CGM metrics, and alternative laboratory markers were all significantly associated with obstetric and neonatal outcomes at 24 weeks' gestation. More outcomes were associated with CGM metrics during the first trimester and with laboratory markers (area under the ROC curve generally <0.7) during the third trimester. Time in range (TIR) (63-140 mg/dL [3.5-7.8 mmol/L]) and time above range (TAR) (>140 mg/dL [>7.8 mmol/L]) were the most consistently predictive CGM metrics. HbA1c was also a consistent predictor of suboptimal pregnancy outcomes. Some alternative laboratory markers showed promise, but overall, they had lower predictive ability than HbA1c. HbA1c is still an important biomarker for obstetric and neonatal outcomes in type 1 diabetes pregnancy. Alternative biochemical markers of glycemia and other CGM metrics did not substantially increase the prediction of pregnancy outcomes compared with widely available HbA1c and increasingly available CGM metrics (TIR and TAR).


Dosage of the smallest chromosome affects both the yeast-hyphal transition and the white-opaque transition of Candida albicans WO-1.
The WO-1 strain of Candida albicans is capable of alternating between two highly distinct yeast cell types termed white and opaque (E. H. A. Rikkerrink, B. B. Magee, and P. T. Magee, J. Bacteriol. 170:895-899, 1988; B. Slutsky, M. Staebell, J. Anderson, L. Risen, M. Pfaller, and D. R. Soll, J. Bacteriol. 169:189-197, 1987). We have isolated WO-1 mutants that show a marked deficiency at being able to switch from the white form to the opaque form under conditions normally favorable for this transition. Pulsed-field electrophoresis demonstrated that one of the initial two spontaneous nonswitching mutants lacked the smallest chromosome that is normally present in WO-1. The availability of a WO-1 derivative whose only functional ADE2 gene is located on this small chromosome made possible, through the induction of chromosome nondisjunction, the isolation of numerous new mutants missing this chromosome as well as mutants containing two copies of the chromosome. Mutants missing the smallest chromosome showed a greatly diminished ability to produce opaque sectors and to produce germ tubes in the presence of human serum. Mutants containing two copies of the small chromosome showed an increased ability to produce germ tubes. These results indicate that this small chromosome carries one or more genes involved in both the white-opaque switch and the yeast-hyphal switch.


A single amino acid substitution in the S1 and S2 Spike protein domains determines the neutralization escape phenotype of SARS-CoV.
In response to SARS-CoV infection, neutralizing antibodies are generated against the Spike (S) protein. Determination of the active regions that allow viral escape from neutralization would enable the use of these antibodies for future passive immunotherapy. We immunized mice with UV-inactivated SARS-CoV to generate three anti-S monoclonal antibodies, and established several neutralization escape mutants with S protein. We identified several amino acid substitutions, including Y442F and V601G in the S1 domain and D757N and A834V in the S2 region. In the presence of each neutralizing antibody, double mutants with substitutions in both domains exhibited a greater growth advantage than those with only one substitution. Importantly, combining two monoclonal antibodies that target different epitopes effected almost complete suppression of wild type virus replication. Thus, for effective passive immunotherapy, it is important to use neutralizing antibodies that recognize both the S1 and S2 regions.


Language and reading abilities of children with autism spectrum disorders and specific language impairment and their first-degree relatives.
Autism spectrum disorder (ASD) and specific language impairment (SLI) are developmental disorders exhibiting language deficits, but it is unclear whether they arise from similar etiologies. Language impairments have been described in family members of children with ASD and SLI, but few studies have quantified them. In this study, we examined IQ, language, and reading abilities of ASD and SLI children and their first-degree relatives to address whether the language difficulties observed in some children with ASD are familial and to better understand the degree of overlap between these disorders and their broader phenotypes. Participants were 52 autistic children, 36 children with SLI, their siblings, and their parents. The ASD group was divided into those with (ALI, n=32) and without (ALN, n=20) language impairment. Relationships between ASD severity and language performance were also examined in the ASD probands. ALI and SLI probands performed similarly on most measures while ALN probands scored higher. ALN and ALI probands' language scores were not related to Autism Diagnostic Interview-Revised and Autism Diagnostic Observation Schedule algorithm scores. SLI relatives scored lowest on all measures, and while scores were not in the impaired range, relatives of ALI children scored lower than relatives of ALN children on some measures, though not those showing highest heritability in SLI. Given that ALI relatives performed better than SLI relatives across the language measures, the hypothesis that ALI and SLI families share similar genetic loading for language is not strongly supported.


The angiogenesis induced by HIV-1 tat protein is mediated by the Flk-1/KDR receptor on vascular endothelial cells.
The HIV-1 Tat protein transactivates HIV, viral and some host cell genes. Tat can be released by infected cells and acts extracellularly in the microenvironment, regulating functions of immunocompetent and mesenchymal cells. One of the most striking effects of Tat is the induction of a functional program in vascular cells related to angiogenesis and inflammation (migration, proliferation and expression of plasminogen activator inhibitor-1 and E selectin). Tat induces growth of Kaposi's sarcoma (KS) spindle cells and is angiogenic in vivo and in transgenic mice10-12. We previously reported that Tat is a direct angiogenic factor and noted the Tat arginine- and lysine-rich sequence is similar to that of other potent angiogenic growth factors, such as vascular endothelial growth factor-A (VEGF-A). It is possible that Tat mimics one of these factors by interacting with its growth factor tyrosine kinase receptor. Here we demonstrate that Tat specifically binds and activates the Flk-1/kinase insert domain receptor (Flk-1/KDR), a VEGF-A tyrosine kinase receptor (for review see ref. 13), and that Tat-induced angiogenesis is blocked by agents blocking the Flk-1/KDR receptor. Endothelial cell stimulation by Tat occurs in the absence of activation of FLT-1, another VEGF-A tyrosine kinase receptor.


A synthetic biosocial model of fertility transition: testing the relative contribution of embodied capital theory, changing cultural norms, and women's labor force participation.
This article presents a biosocial model of fertility decline, which integrates ecological-economic and informational-cultural hypotheses of fertility transition in a unified theoretical framework. The model is then applied to empirical data collected among 500 women from San Borja, Bolivia, a population undergoing fertility transition. Using a combination of event history analysis, multiple regression, and structural equation modeling, we examine the pathways by which education responds to birth cohort, parental education and network ties, and how age at first birth and total fertility, in turn, respond to birth cohort, social network ties, education, expectations about parental investment, work, and contraceptive use. We find that in addition to secular trends in education, respondent's education is associated with the education of parents, the investment she received from them, and the education of older siblings. Total fertility has dropped over time, partly in response to increased education; moreover, the behavior of other women in a woman's social network predicts both initiation of reproduction and total fertility, while expected parental investment in offspring negatively predicts total fertility. Involvement in paid work that is incompatible with childcare is associated with a later age of first reproduction, but not subsequent fertility. Contraceptive use partially mediates the effect of education and birth cohort on total fertility, but is not a mediator of the effect of social network or expected parental investment on total fertility. Overall, the empirical results provide support for a biosocial model of fertility decline, particularly the embodied capital and cultural pathways.


Risk stratification in the patient with non ST segment elevation acute coronary syndrome.
Patients hospitalized with unstable angina (UA) or with a non-ST segment elevation myocardial infarct (NSTEMI) are at increased risk of suffering refractory angina, recurrent myocardial infarct (MI), and death. These patients need to be evaluated more aggressively. According to the last published guidelines (2002) of UA/NSTEMI by the ACC/AHA Task Force, these patients should be categorized in a risk scale as: low, intermediate or high. This should be done in the initial evaluation, which includes: medical history, physical exam, an electrocardiogram (ECG) and cardiac markers. The TIMI risk score should also be used as complementary in this risk assessment. High risk patients, without contraindications, should be managed more aggressively with coronary angiography. On the other end, low risk patients, and some intermediate, may be evaluated more conservatively with early non-invasive studies for further assessment of ischemia and prognosis.


Triclosan enriches for Dehalococcoides-like Chloroflexi in anaerobic soil at environmentally relevant concentrations.
Triclosan is an antimicrobial agent that is discharged to soils with land-applied wastewater biosolids, is persistent under anaerobic conditions, and yet its impact on anaerobic microbial communities in soils is largely unknown. We hypothesized that triclosan enriches for Dehalococcoides-like Chloroflexi because these bacteria respire organochlorides and are likely less sensitive, relative to other bacteria, to the antimicrobial effects of triclosan. Triplicate anaerobic soil microcosms were seeded with agricultural soil, which was not previously exposed to triclosan, and were amended with 1 mg kg(-1) of triclosan. Triplicate control microcosms did not receive triclosan, and the experiment was run for 618 days. The overall bacterial community (assessed by automated ribosomal intergenic spacer analysis and denaturing gradient gel electrophoresis) was not impacted by triclosan; however, the abundance of Dehalococcoides-like Chloroflexi 16S rRNA genes (determined by qPCR) increased 20-fold with triclosan amendment compared with a fivefold increase without triclosan. This work demonstrates that triclosan impacts anaerobic soil communities at environmentally relevant levels.


Snapshot of Quantiferon TB gold testing in Northern Mexico.
Most people infected with Mycobacterium tuberculosis have an asymptomatic condition named latent tuberculosis. These people do not have bacilli in the corporal secretions and are hard to diagnose by conventional laboratory tests. Diagnosis of latent tuberculosis infection (LTBI) in México is based on the tuberculin skin test (TST). This test has disadvantages, principally because the vaccine containing the Bacille Calmette-Guérin (BCG) is applied to 99% of this population and causes false positive TST outcomes. Recently, interferon-gamma release assays (IGRA) have been demonstrated to be a good test to detect latent tuberculosis with equal or better sensitivity to TST and without interference from BCG. However, in México the IGRA are an uncommon test due to the higher cost compared to TST. The main objective of this work was demonstrate the potential utility of the Quantiferon TB(®) gold in tube (QTB(®)-GIT) test to detect latent TB in a population from northern México. Samples from 106 subjects with close contact, or without contact, with actively infected TB patients were tested to detect LTBI. Our results show a significant difference between individuals in close contact with active TB patients (39.7%) compared to those without contact (3.2%), p < 0.01. The concordance between TST and QTB(®)-GIT was poor (κ = 0.31). Our preliminary results show that the QTB(®)-GIT has better capacity than TST to detect latent tuberculosis infection.


Clinical, economic, and humanistic burden of asthma in Canada: a systematic review.
Asthma, one of the most common chronic respiratory diseases, affects about 3 million Canadians. The objective of this study is to provide a comprehensive evaluation of the published literature that reports on the clinical, economic, and humanistic burden of asthma in Canada. A search of the PubMed, EMBASE, and EMCare databases was conducted to identify original research published between 2000 and 2011 on the burden of asthma in Canada. Controlled vocabulary with "asthma" as the main search concept was used. Searches were limited to articles written in English, involving human subjects and restricted to Canada. Articles were selected for inclusion based on predefined criteria like appropriate study design, disease state, and outcome measures. Key data elements, including year and type of research, number of study subjects, characteristics of study population, outcomes evaluated, results, and overall conclusions of the study, were abstracted and tabulated. Thirty-three of the 570 articles identified by the clinical and economic burden literature searches and 14 of the 309 articles identified by the humanistic burden literature searches met the requirements for inclusion in this review. The included studies highlighted the significant clinical burden of asthma and show high rates of healthcare resource utilization among asthma patients (hospitalizations, ED, physician visits, and prescription medication use). The economic burden is also high, with direct costs ranging from an average annual cost of $366 to $647 per patient and a total annual population-level cost ranging from ~ $46 million in British Columbia to ~ $141 million in Ontario. Indirect costs due to time loss from work, productivity loss, and functional impairment increase the overall burden. Although there is limited research on the humanistic burden of asthma, studies show a high (31%-50%) prevalence of psychological distress and diminished QoL among asthma patients relative to subjects without asthma. As new therapies for asthma become available, economic evaluations and assessment of clinical and humanistic burden will become increasingly important. This report provides a comprehensive resource for health technology assessment that will assist decision making on asthma treatment selection and management guidelines in Canada.


Dietary total antioxidant capacity is associated with leukocyte telomere length in a children and adolescent population.
Oxidative stress and inflammation seem to be potential underlying mechanisms for telomere attrition. A lack of specific antioxidants is believed to increase free radical damage and a greater risk for telomere shortening. Our aim was to evaluate the relationship between diet and leukocyte telomere length in a cross-sectional study of children and adolescents. We hypothesized that dietary total antioxidant capacity would be positively associated with telomere length. Telomere length was measured by quantitative real-time polymerase chain reaction in 287 participants (55% males, 6-18 years), who were randomly selected from the GENOI study. A positive correlation between dietary total antioxidant capacity and telomere length (r = 0.157, p = 0.007) was found after adjustment for age and energy intake. However, higher white bread consumption was associated with shorter telomeres (β = -0.204, p = 0.002) in fully-adjusted models. Interestingly, those individuals who had simultaneously higher dietary total antioxidant capacity and lower white bread consumption significantly presented the longest telomeres. Moreover, the multivariable-adjusted odds ratio for very short telomeres was 0.30 for dietary total antioxidant capacity (p = 0.023) and 1.37 for white bread (p = 0.025). It was concluded that longer telomeres were associated with higher dietary total antioxidant capacity and lower white bread consumption in Spanish children and adolescents. These findings might open a new line of investigation about the potential role of an antioxidant diet in maintaining telomere length.


Shared T cell recognition sites on human histocompatibility leukocyte antigen class II molecules of patients with seropositive rheumatoid arthritis.
Seropositive rheumatoid arthritis (RA) in adult and juvenile patients is associated with the serologic marker HLA-DR4. This association is incomplete; about one-third of the patients lack the disease-associated HLA-DR4 haplotype. The main biological function of class II molecules is to restrict the recognition of antigen by T lymphocytes. We therefore tested the hypothesis that patients with seropositive RA share T cell recognition sites for an unknown antigen and that such T cell "epitopes" are not identified by conventional serologic typing. We generated alloreactive human T cell clones by stimulating peripheral blood lymphocytes of normal donors against a lymphoblastoid cell line from a juvenile patient with seropositive RA. A panel of clones that recognized only HLA-Dw14 cells on a panel of homozygous typing cells was used to analyze class II molecules of adult patients with seropositive RA. By inhibition studies using monoclonal antibodies, the epitopes recognized by the different clones could be further characterized and assigned either to DR- or to DQ-encoded cell surface products. By using four different clones, it was possible to identify Dw14-associated T cell epitopes on all seropositive rheumatoid patients tested who typed HLA-DR4-positive and also on all eight DR4-negative patients tested. Approximately one-half of nonrheumatoid DR4-positive donors carried one or more determinants recognized by these clones; the expression of these allodeterminants in DR4-negative nonrheumatoid patients was rare (less than 10%). Thus, alloreactive human T cell clones are powerful tools to define T cell recognition sites on class II molecules that are not identified by conventional typing. Using T cell clones with specificities for determinants expressed on Dw14 homozygous typing lines, we were able to demonstrate shared epitopes on cells of all patients tested with seropositive RA irrespective of their HLA-D or HLA-DR type. These data suggest that major histocompatibility complex class II antigens of RA patients might be much more homogeneous than demonstrated by the incomplete HLA-DR4 association.


Are the current thresholds, indicators, and time window for cold warning effective enough to protect cardiovascular health?
More extreme cold weather and larger weather variations have raised concerns regarding their effects on public health. Although prior studies assessed the effects of cold air temperature on health, especially mortality, limited studies evaluated wind chill temperatures on morbidity, and health effects under the current cold warning threshold. This study identified the thresholds, lag periods, and best indicators of extreme cold on cardiovascular disease (CVD) by comparing effects of wind chill temperatures and cold air temperatures on CVD emergency department (ED) visits in winter and winter transition months. Information was collected on 662,625 CVD ED visits from statewide hospital discharge dataset in New York State. Meteorological factors, including air temperature, wind speed, and barometric pressure were collected from National Oceanic and Atmospheric Administration. A case-crossover approach was used to assess the extreme cold-CVD relationship in winter (December-February) and transition months (November and March) after controlling for PM2.5. Conditional logistic regression models were employed to analyze the association between cold weather factors and CVD ED visits. We observed CVD effects occurred when wind chill temperatures were as high as -3.8 °C (25 °F), warmer than current wind chill warning standard (≤-28.8 °C or ≤-20 °F). Wind chill temperature was a more sensitive indicator of CVD ED visits during winter with temperatures ≤ -3.8 °C (25 °F) with delay effect (lag 6); however, air temperature was better during transition months for temperatures ≤ 7.2 °C (45 °F) at earlier lag days (1-3). Among all CVD subtypes, hypertension ED visit had the strongest negative association with both wind chill temperature and air temperature. This study recommends modifying the current cold warning temperature threshold given larger proportions of CVD cases are occurring at considerably higher temperatures than the current criteria. We also recommend issuing cold warnings in winter transitional months.


25th anniversary article: organic photovoltaic modules and biopolymer supercapacitors for supply of renewable electricity: a perspective from Africa.
The role of materials in civilization is well demonstrated over the centuries and millennia, as materials have come to serve as the classifier of stages of civilization. With the advent of materials science, this relation has become even more pronounced. The pivotal role of advanced materials in industrial economies has not yet been matched by the influence of advanced materials during the transition from agricultural to modern societies. The role of advanced materials in poverty eradication can be very large, in particular if new trajectories of social and economic development become possible. This is the topic of this essay, different in format from the traditional scientific review, as we try to encompass not only two infant technologies of solar energy conversion and storage by means of organic materials, but also the social conditions for introduction of the technologies. The development of organic-based photovoltaic energy conversion has been rapid, and promises to deliver new alternatives to well-established silicon photovoltaics. Our recent development of organic biopolymer composite electrodes opens avenues towards the use of renewable materials in the construction of wooden batteries or supercapacitors for charge storage. Combining these new elements may give different conditions for introduction of energy technology in areas now lacking electrical grids, but having sufficient solar energy inputs. These areas are found close to the equator, and include some of the poorest regions on earth.


The place and practices of well-being in local governance.
The concept of well-being has become prominent within national policy goals in the UK since the end of the 1990s. However, the concept of well-being remains ill defined, an instability that is increasingly understood as problematic to policy making. We engage with this terminological instability through an exploration of how the concept of well-being is practised discursively in local governance and critically examine the place of the concept in local policy making. In contrast to the current enthusiasm to define and measure well-being, we argue that the conceptual instability has inherent value for local governance. The concept of well-being is practised through a number of potentially conflicting discourses, but it is exactly this conceptual instability that enables a local negotiation and combination of alternative policy frameworks for local place-shaping strategies. As such, well-being not only is an overarching goal of governance but also contributes to the dynamics of the policy process.


Sharing Structure and Function in Biological Design with SBOL 2.0.
The Synthetic Biology Open Language (SBOL) is a standard that enables collaborative engineering of biological systems across different institutions and tools. SBOL is developed through careful consideration of recent synthetic biology trends, real use cases, and consensus among leading researchers in the field and members of commercial biotechnology enterprises. We demonstrate and discuss how a set of SBOL-enabled software tools can form an integrated, cross-organizational workflow to recapitulate the design of one of the largest published genetic circuits to date, a 4-input AND sensor. This design encompasses the structural components of the system, such as its DNA, RNA, small molecules, and proteins, as well as the interactions between these components that determine the system's behavior/function. The demonstrated workflow and resulting circuit design illustrate the utility of SBOL 2.0 in automating the exchange of structural and functional specifications for genetic parts, devices, and the biological systems in which they operate.


Multinozzle emitter arrays for nanoelectrospray mass spectrometry.
Mass spectrometry (MS) is the enabling technology for proteomics and metabolomics. However, dramatic improvements in both sensitivity and throughput are still required to achieve routine MS-based single cell proteomics and metabolomics. Here, we report the silicon-based monolithic multinozzle emitter array (MEA) and demonstrate its proof-of-principle applications in high-sensitivity and high-throughput nanoelectrospray mass spectrometry. Our MEA consists of 96 identical 10-nozzle emitters in a circular array on a 3 in. silicon chip. The geometry and configuration of the emitters, the dimension and number of the nozzles, and the micropillar arrays embedded in the main channel can be systematically and precisely controlled during the microfabrication process. Combining electrostatic simulation and experimental testing, we demonstrated that sharpened-end geometry at the stem of the individual multinozzle emitter significantly enhanced the electric fields at its protruding nozzle tips, enabling sequential nanoelectrospray for the high-density emitter array. We showed that electrospray current of the multinozzle emitter at a given total flow rate was approximately proportional to the square root of the number of its spraying-nozzles, suggesting the capability of high MS sensitivity for multinozzle emitters. Using a conventional Z-spray mass spectrometer, we demonstrated reproducible MS detection of peptides and proteins for serial MEA emitters, achieving sensitivity and stability comparable to the commercial capillary emitters. Our robust silicon-based MEA chip opens up the possibility of a fully integrated microfluidic system for ultrahigh-sensitivity and ultrahigh-throughput proteomics and metabolomics.


Activin A is an anticatabolic autocrine cytokine in articular cartilage whose production is controlled by fibroblast growth factor 2 and NF-kappaB.
Proteomic analysis has previously shown that activin A, a member of the transforming growth factor beta family, is produced by human articular cartilage. This study was undertaken to investigate whether activin A affects cartilage matrix catabolism and how its production is regulated. The effect of exogenous activin A on interleukin-1-induced aggrecanase-generated neoepitope production was assessed by Western blotting, using medium from human cartilage explants. Levels of activin A production were determined by enzyme-linked immunosorbent assay. For genes of interest, messenger RNA (mRNA) induction in cartilage explants or primary chondrocyte monolayers was assessed by reverse transcriptase-polymerase chain reaction. Activin A activity in cartilage explant medium was measured by incubating it with human dermal fibroblasts and determining the increase in phospho-Smad2 by Western blotting. Activin A (1-10 ng/ml) suppressed aggrecanase-mediated cleavage of aggrecan in human articular cartilage. Activin A mRNA and protein secretion were induced by dissection and culture of human or porcine articular cartilage. This activin A was biologically active. Its production was due to an active cellular process and was enhanced in osteoarthritic (OA) tissue. Activin A production on dissection was reduced by 80% by the fibroblast growth factor (FGF) receptor inhibitor PD173074 and by 70% by the IKK inhibitor BMS345541. Activin A is potentially an anticatabolic molecule in articular cartilage. Its expression is induced by wounding in an FGF-2- and NF-kappaB-dependent manner. OA cartilage produced more activin A than did normal cartilage in vitro.


[Anesthetic induction after treated hemorrhagic shock: experimental study comparing ketamine and etomidate.].
Bleeding causing hemorrhagic shock usually requires surgical treatment under general anesthesia. Anesthetic drugs may further compromise hemodynamics. The objective was to compare the hemodynamic effects of ketamine and etomidate during anesthetic induction in dogs submitted to an experimental model of hemorrhagic shock and resuscitation. Thirty-two mongrel dogs were submitted to a pressure-controlled hemorrhagic shock, resuscitation and anesthetic induction model. After achieving the target pressure of 40 mmHg, they were randomly assigned in two groups according to the resuscitation fluid to be used: NaCl 0.9% (32 mL.kg-1) and NaCl 7.5% (4 mL.kg-1). After volume infusion, these groups were reassigned according to anesthetic drug used: GI) NaCl 0.9% and ketamine; GII) NaCl 7.5% and ketamine; GIII) NaCl 0.9% and etomidate; and GIV) NaCl 7.5% and etomidate. Hemodynamic measurements were obtained at five moments: (M0) baseline; (M1) after bleeding to shock; (M2) after volume expansion; (M3) 5 minutes after anesthetic induction; (M4) 15 minutes after anesthetic induction. Statistical analysis was performed using Student t test and two way ANOVA. Value of p lower than 0.05, was considered significant. After shock, both solutions restored hemodynamics to baseline values. Independently of anesthetic agent or expansion solution used, mean arterial pressure remained unaltered for all groups after induction. Central venous pressure, heart rate, pulmonary capillary wedge pressure and pulmonary vascular resistance index increased significantly after ketamine infusion. Cardiac index, systemic vascular resistance index and oxygen transport variables remained stable in all groups. Etomidate or ketamine were able to maintain hemodynamic stability in dogs undergoing severe hemorrhagic shock treated with NaCl 0.9% or NaCl 7.5%.


Genetic contribution to rate of change in functional abilities among Danish twins aged 75 years or more.
In a previous cross-sectional study of twins, the authors found evidence of a substantial genetic influence on functional abilities among elderly women. It has been suggested that rate of change in functional abilities over time could underlie such findings and that rate-of-change phenotypes may have an even larger genetic component than "level" phenotypes (e.g., functional abilities per se). If so, rate-of-change phenotypes could be more powerful than level phenotypes in studies aimed at identifying specific polymorphisms of importance for aging. In 1995, the authors assessed a population-based sample of 2,401 Danish twins aged 75 years or more. The survivors were recontacted after 2 years and again after 4 years. Consistent mean-level declines, high within-person correlations over time, and substantial heritability in the female sample were observed for functional abilities. Nonetheless, structural-equation analyses revealed only a very modest and nonsignificant heritability for rate of change in functional abilities: 16% (95% confidence interval: 0, 35) for women and 9% (95% confidence interval: 0, 44) for men. This study had a large initial sample size, high participation rates, and a valid and reliable measure of rate of change in a phenotype that had previously shown substantial heritability in cross-sectional analyses in the same twin population. Still, the present study revealed only a modest and nonsignificant genetic influence on rate of change, which suggests that detection of polymorphisms influencing rate of change in functional abilities among the elderly may prove to be difficult.


Multi-dimensional time-correlated single photon counting (TCSPC) fluorescence lifetime imaging microscopy (FLIM) to detect FRET in cells.
We present a novel, multi-dimensional, time-correlated single photon counting (TCSPC) technique to perform fluorescence lifetime imaging with a laser-scanning microscope operated at a pixel dwell-time in the microsecond range. The unsurpassed temporal accuracy of this approach combined with a high detection efficiency was applied to measure the fluorescent lifetimes of enhanced cyan fluorescent protein (ECFP) in isolation and in tandem with EYFP (enhanced yellow fluorescent protein). This technique enables multi-exponential decay analysis in a scanning microscope with high intrinsic time resolution, accuracy and counting efficiency, particularly at the low excitation levels required to maintain cell viability and avoid photobleaching. Using a construct encoding the two fluorescent proteins separated by a fixed-distance amino acid spacer, we were able to measure the fluorescence resonance energy transfer (FRET) efficiency determined by the interchromophore distance. These data revealed that ECFP exhibits complex exponential fluorescence decays under both FRET and non-FRET conditions, as previously reported. Two approaches to calculate the distance between donor and acceptor from the lifetime delivered values within a 10% error range. To confirm that this method can be used also to quantify intermolecular FRET, we labelled cultured neurones with the styryl dye FM1-43, quantified the fluorescence lifetime, then quenched its fluorescence using FM4-64, an efficient energy acceptor for FM1-43 emission. These experiments confirmed directly for the first time that FRET occurs between these two chromophores, characterized the lifetimes of these probes, determined the interchromophore distance in the plasma membrane and provided high-resolution two-dimensional images of lifetime distributions in living neurones.


Beta-glucan reflects liver injury after preservation and transplantation in dogs.
Graft failure and extrahepatic organ complications, which frequently develop after transplantation, may be related to inflammatory mediators stimulated by endotoxin (ET). The role of endotoxemia after liver transplantation is controversial and may depend upon differences in the ET assay method used in the various contradicting studies. While the standard Limulus amebocyte lysate (LAL) is reactive for ET and beta-glucan, a novel turbidimetric assay method enables separate determinations of ET and beta-glucan. Beagle dogs undergoing orthotopic liver transplantation were divided into two groups. In Group I (n = 6) the grafts were transplanted immediately and in Group II (n = 6) grafts were preserved for 48 h in University of Wisconsin (UW) solution. Animals received cyclosporine immunosuppression and were followed for 14 days. Daily measurements of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and lactate dehydrogenase (LDH) were performed. Samples for ET and beta-glucan measurement were collected serially and processed using the turbidimetric assay method. While no graft failure was seen in Group I, three of six Group II animals died from graft failure within 1 day after transplantation. Preservation and reperfusion injury was much more severe in the Group II grafts than in Group I grafts. While endotoxemia could not be detected, postoperative beta-glucan levels (undetectable pretransplant) were seen in both groups. Beta-glucan levels were much higher in Group II grafts than in Group I grafts, and correlated with the severity of liver damage. In conclusion, this study shows that beta-glucan, instead of ET, appears during the early posttransplant period. We believe that posttransplant elevation of beta-glucan is related to liver damage, especially endothelial damage by preservation and reperfusion.


An essential gene pair in Saccharomyces cerevisiae with a potential role in mating.
In the yeast Saccharomyces cerevisiae, the signal generated by extracellular pheromone is transmitted through the beta and gamma subunits of a trimeric G-protein to downstream signaling molecules that mediate the cellular responses associated with mating. To isolate potential downstream signaling components, a yeast genomic library on a multicopy plasmid was screened for genes that increased the mating efficiency of a strain containing a temperature-sensitive G beta subunit mutation. Overexpression of STE5, STE18 (which encodes the G gamma subunit), and a previously unidentified gene, termed SSF1, partially suppressed the mating defect of a G beta mutant. Hybridization of yeast genomic DNA with an SSF1 probe revealed a closely related homolog, termed SSF2, which was isolated and also found to test positively in the assay for suppression. Null mutations in either SSF1 or SSF2 had no obvious phenotype, but disruption of both genes was lethal. Depletion of SSF gene products from growing cultures caused both an arrest of cell division and a significant decrease in the ability of cells to mate. Because mating efficiency was increased by extra copies of the SSF genes and decreased by elimination of the gene products, it is likely that these genes play a role in mating as well as in an essential function.


Preparation of difluoromethylthioethers through difluoromethylation of disulfides using TMS-CF2H.
We report an operationally simple, metal-free approach for the late-stage introduction of the important lipophilic hydrogen-bond donor motif, SCF2H. This reaction converts diaryl- and dialkyl-disulfides into the corresponding aryl/alkyl-SCF2H through the nucleophilic transfer of a difluoromethyl group with good functional group tolerance. This method is notable for its use of commercially available TMSCF2H, and does not rely on the need for handling of sensitive metal complexes.


Neglected side effects after radical prostatectomy: a systematic review.
A series of previously neglected sexually related side effects to radical prostatectomy (RP) has been identified over the recent years. These include orgasm-associated incontinence (OAI), urinary incontinence in relation to sexual stimulation (UISS), altered perception of orgasm, orgasm-associated pain (OAP), penile shortening (PS), and penile deformity. The aim of this article is to conduct a systematic review of the literature regarding the above-mentioned side effects. A predefined search strategy was applied in a thorough search of Medline, Web of science, and the online Cochrane library. The PRISMA guidelines for systematic reviews were followed, and protocol as well as search strategies was registered at http://www.crd.york.ac.uk/Prospero/ (RN: CRD42012003165). The main outcome measure was incidence rates for the relevant side effects. A total of 43 articles were included. OAI and UISS are experienced by 20-93% of RP patients at least a few times after surgery. Although these issues are associated to postoperative daytime incontinence, previous transurethral resection of the prostate (TURP) is the only known predicting factor. Alterations of orgasmic function are experienced by approximately 80% after RP. Erectile dysfunction seems to play an important role in waning orgasmic function. OAP is only experienced by a subset of the patients with reported rates varying between 3% and 19%. Sparing of the tips of the seminal vesicles has been shown to double the risk of OAP. PS occurs in 15-68% of RP patients. Nerve sparing and preservation of erectile function may help preserve penile length. With regard to all side effects, studies indicate that they are reduced over time. The sexually related side effects summarized in this review are common after RP. Meanwhile, it is difficult to predict which patients are at risk. Daytime incontinence, previous TURP, a lack of nerve sparing, and erectile dysfunction are all associated with the above-mentioned sexually related side effects.


Direct sequencing of small genomes on the Pacific Biosciences RS without library preparation.
We have developed a sequencing method on the Pacific Biosciences RS sequencer (the PacBio) for small DNA molecules that avoids the need for a standard library preparation. To date this approach has been applied toward sequencing single-stranded and double-stranded viral genomes, bacterial plasmids, plasmid vector models for DNA-modification analysis, and linear DNA fragments covering an entire bacterial genome. Using direct sequencing it is possible to generate sequence data from as little as 1 ng of DNA, offering a significant advantage over current protocols which typically require 400-500 ng of sheared DNA for the library preparation.


Importance of main pancreatic duct dilatation in IPMN undergoing surveillance.
The association between risk of pancreatic cancer and a dilated main pancreatic duct (MPD) in intraductal papillary mucinous neoplasm (IPMN) is debated. The aim of this study was to assess the role of MPD size in predicting pancreatic cancer in resected IPMNs and those kept under surveillance. All patients with IPMN referred to the Pancreas Institute, University of Verona Hospital Trust, from 2006 to 2016 were included. The primary endpoint was the occurrence of malignancy detected at surgery or during follow-up. The final cohort consisted of 1688 patients with a median follow-up of 60 months. Main pancreatic duct dilatation was associated with other features of malignancy in both the resected and surveillance groups. In patients who underwent resection, only a MPD of at least 10 mm was an independent predictor of malignancy. In patients kept under surveillance, MPD dilatation was not associated with malignancy. Fifteen of 71 patients (21 per cent) with malignancy in the resection cohort had a dilated MPD alone, whereas only one of 30 (3 per cent) under surveillance with MPD dilatation alone developed malignancy. Patients with a dilated MPD and other worrisome features had an increased 5-year cumulative incidence of malignancy compared with those with a non-dilated duct (11 versus 1·2 per cent; P < 0·001); however, the risk of malignancy was not significantly increased in patients with a dilated MPD alone (4 versus 1·2 per cent; P = 0·448). In patients under surveillance, a dilated MPD alone was not associated with an increased incidence of malignancy in IPMN.


Complex drug use patterns and associated HIV transmission risk behaviors in an Internet sample of U.S. men who have sex with men.
Although the relationship between drug use and HIV risk among men who have sex with men (MSM) is well described, relatively few studies have employed empirical methods to assess underlying classes of drug use that may better predict the risk of HIV or sexually transmitted infections (STIs) among MSM. The aim of this study was to determine whether latent class analysis (LCA) would identify underlying drug classes reported prior to sex, as well as predict unprotected anal intercourse (UAI) in the last sexual encounter among MSM. From 2004 to 2005, an anonymous online survey was conducted among 8,717 sexually active MSM recruited from gay-affiliated U.S. websites. LCA clustered participants into six distinct drug use classes based on the specific types and number of drugs used: (1) low/no drug use, (2) recreational drug use, (3) poppers with prescription erectile dysfunction (ED) drug use, (4) poppers with both prescription and non-prescription ED drug use, (5) recreational, club, and ED drug use, and (6) high polydrug use. Compared with men in Class 1, men in the highest drug use class were 4.84 times more likely to report UAI in their last sexual encounter and 3.78 times more likely to report an STI in the past year (both ps < .001). Younger MSM aged 18-29 were significantly more likely to report an STI than men aged 50 and above (p < .001). There is a need to better understand the complex relationship between a diverse set of drugs used among MSM and how polydrug use impacts sexual negotiation over time.


Sodium-ion storage properties of nickel sulfide hollow nanospheres/reduced graphene oxide composite powders prepared by a spray drying process and the nanoscale Kirkendall effect.
Spray-drying and the nanoscale Kirkendall diffusion process are used to prepare nickel sulfide hollow nanospheres/reduced graphene oxide (rGO) composite powders with excellent Na-ion storage properties. Metallic Ni nanopowder-decorated rGO powders, formed as intermediate products, are transformed into composite powders of nickel sulfide hollow nanospheres/rGO with mixed crystal structures of Ni3S2 and Ni9S8 phases by the sulfidation process under H2S gas. Nickel sulfide/rGO composite powders with the main crystal structure of Ni3S2 are also prepared as comparison samples by the direct sulfidation of nickel acetate-graphene oxide (GO) composite powders obtained by spray-drying. In electrochemical properties, the discharge capacities at the 150(th) cycle of the nickel sulfide/rGO composite powders prepared by sulfidation of the Ni/rGO composite and nickel acetate/GO composite powders at a current density of 0.3 A g(-1) are 449 and 363 mA h g(-1), respectively; their capacity retentions, calculated from the tenth cycle, are 100 and 87%. The nickel sulfide hollow nanospheres/rGO composite powders possess structural stability over repeated Na-ion insertion and extraction processes, and also show excellent rate performance for Na-ion storage.


The role and advances of immunomodulator therapy for inflammatory bowel disease.
Immune modulating drugs such as thiopurines (azathioprine and 6-mercaptopurine) and methotrexate has been a mainstay for treatment of inflammatory bowel disease (IBD) for decades. However, despite widely used in IBD, questions still remain concerning the most rational treatment regimens of these agents. Results from a range of recent studies necessitate increased awareness on how to best use these potent drugs in the clinic. As controversy still remains regarding the most appropriate use of immunomodulators, this review is based on scrutinizing the current literature, with emphasis on randomized controlled trials and Cochrane reviews, focusing on aspects that can lead to optimal and evidence-based thiopurine and methotrexate treatment strategies in IBD.


Effect of the metalloproteinase inhibitor batimastat in the systemic toxicity induced by Bothrops asper snake venom: understanding the role of metalloproteinases in envenomation.
The peptidomimetic hydroxamate metalloproteinase inhibitor batimastat (BB-94) was assessed for its ability to neutralize the systemic effects (lethality, hemorrhage and coagulopathy) induced by the venom of Bothrops asper, the most important snake from a medical standpoint in Central America. Batimastat inhibited lethality when a venom challenge dose of two LD(50)s was used by intraperitoneal and intravenous routes, with ED(50)s of 250 and 22 microM, respectively. With a challenge dose of three LD(50)s, lethality was not abrogated, but a conspicuous and dose-dependent delay in the time of death was observed in mice injected with mixtures of venom plus batimastat. Upon incubation with 500 microM batimastat, venom LD(50) increased 2.86-fold (intraperitoneal route) and 2.37-fold (intravenous route), when compared with LD(50) of venom alone. Batimastat also inhibited the hemorrhagic effect induced by venom in the lungs after intravenous injection. Moreover, batimastat exerted a significant inhibition of in vitro coagulant and in vivo defibrinogenating effects of venom, evidencing that metalloproteinases play a key role in the coagulopathy characteristic of B. asper envenomation. The remaining uninhibited coagulant effect is due to serine proteinases, i.e. thrombin-like enzymes, since this effect was completely abrogated by the combination of batimastat and PMSF. Our results stress the view that metalloproteinases play a relevant role in the systemic pathophysiology of B. asper envenomation and that metalloproteinase inhibitors may become a therapeutic alternative in this pathology.


Reassortment of American and Eurasian genes in an influenza A virus isolated from a great black-backed gull (Larus marinus), a species demonstrated to move between these regions.
The primary hosts for influenza A viruses are waterfowl, although gulls and shorebirds are also important in global avian influenza dynamics. Avian influenza virus genes are separated phylogenetically into two geographic clades, American and Eurasian, which is caused by the geographic separation of the host species between these two regions. We surveyed a gregarious and cosmopolitan species, the Great Black-backed Gull (Larus marinus), in Newfoundland, Canada, for the presence of avian influenza viruses. We have isolated and determined the complete genome sequence of an H13N2 virus, A/Great Black-backed Gull/Newfoundland/296/2008(H13N2), from one of these birds. Phylogenetic analysis revealed that this virus contained two genes in the American gull clade (PB1, HA), two genes in the American avian clade (PA, NA), and four genes in the Eurasian gull clade (PB2, NP, M, NS). We analyzed bird band recovery information and found the first evidence of trans-Atlantic migration from Newfoundland to Europe (UK, Spain and Portugal) for this species. Thus, great black-backed gulls could be important for movement of avian influenza viruses across the Atlantic Ocean and within North America.


Recent research on 3-phenyllactic acid, a broad-spectrum antimicrobial compound.
3-Phenyllactic acid (PLA), which is an organic acid widely existing in honey and lactic acid bacteria fermented food, can be produced by many microorganisms, especially lactic acid bacteria. It was proved as an ideal antimicrobial compound with broad and effective antimicrobial activity against both bacteria and fungi. In addition, it could be used as feed additives to replace antibiotics in livestock feeds. This article presented a review of recent studies on the existing resource, antimicrobial activity, and measurement of PLA. In addition, microorganism strains and dehydrogenases producing PLA were reviewed in detail, the metabolic pathway and regulation of PLA synthesis in LAB strains were discussed, and high-level bioproduction of PLA by microorganism fermentation was also summarized.


The role of mechanical forces on the patterning of the avian feather-bearing skin: A biomechanical analysis of the integumentary musculature in birds.
The integumentary musculature of birds consists of three distinct components. The smooth musculature comprises feather and apterial muscles, which form a continuous musculo-elastic layer within the dermis. The feather muscles, which consistently include at least erectors and depressors, interconnect contour feathers within pterylae (i.e., feather tracts) along gridlines that are oriented diagonally to the longitudinal and transverse axes of the body. The apterial muscles interconnect pterylae by attaching to the contour feathers along their peripheries. The striated musculature is composed of individual subcutaneous muscles, most of which attach to contour feathers along the caudal periphery of pterylae A new integrative functional analysis of the integumentary musculature proposes how apterial muscles stabilize the pterylae and modulate the tension of the musculo-elastic layer, and how subcutaneous muscles provide the initial stimulus for erector muscles being able to ruffle the contour feathers within pterylae. It also shows how the arrangement of the contour feathers and integumentary muscles reflects the stresses and strains that act on the avian skin. These mechanical forces are in effect not only in the adult, especially during flight, but may also be active during feather morphogenesis. The avian integument with its complex structural organization may, therefore, represent an excellent model for analyzing the nature of interactions between the environment and genetic material. The predictions of our model are testable, and our study demonstrates the relevance of integrated analyses of complex organs as mechanically coherent systems for evolutionary and developmental biology.


Spatial clustering of high load ocular Chlamydia trachomatis infection in trachoma: a cross-sectional population-based study.
Chlamydia trachomatis (Ct) is the most common cause of bacterial sexually transmitted infection and infectious cause of blindness (trachoma) worldwide. Understanding the spatial distribution of Ct infection may enable us to identify populations at risk and improve our understanding of Ct transmission. In this study, we sought to investigate the spatial distribution of Ct infection and the clinical features associated with high Ct load in trachoma-endemic communities on the Bijagós Archipelago (Guinea Bissau). We collected 1507 conjunctival samples and corresponding detailed clinical data during a cross-sectional population-based geospatially representative trachoma survey. We used droplet digital PCR to estimate Ct load on conjunctival swabs. Geostatistical tools were used to investigate clustering of ocular Ct infections. Spatial clusters (independent of age and gender) of individuals with high Ct loads were identified using local indicators of spatial association. We did not detect clustering of individuals with low load infections. These data suggest that infections with high bacterial load may be important in Ct transmission. These geospatial tools may be useful in the study of ocular Ct transmission dynamics and as part of trachoma surveillance post-treatment, to identify clusters of infection and thresholds of Ct load that may be important foci of re-emergent infection in communities.


Binding of dorzolamide and its metabolite, N-deethylated dorzolamide, to human erythrocytes in vitro.
Dorzolamide, previously known as MK-507, is a novel topical carbonic anhydrase (CA) inhibitor. The uptake and binding of dorzolamide and its N-deethylated metabolite to human erythrocytes were studied in vitro. Dorzolamide and N-deethylated dorzolamide were preferentially taken up by the erythrocytes, and the uptake of dorzolamide by erythrocytes was found to be much faster than that of N-deethylated dorzolamide. When 20 or 200 microM dorzolamide was incubated with human erythrocytes, 98% or 71% of the drug was taken up by the erythrocytes, respectively. Similarly, using 20 or 200 microM N-deethylated dorzolamide, 99.7% or 75% of the drug was taken up by the erythrocytes, respectively. These results indicate that human erythrocytes contain proteins that bind to dorzolamide and N-deethylated dorzolamide, and the binding of these proteins is saturable. The results of the in vitro binding study suggest that presence of at least three kinds of binding site for dorzolamide in human erythrocytes. One of the binding sites for dorzolamide was characterized by extremely high affinity (Kd = 0.0011 microM) and low capacity (Bmax = 16.1 microM), corresponding to CA-II. Another binding site was characterized by low affinity (Kd = 2.8 microM) and high capacity (Bmax = 117.1 microM), corresponding to CA-I. The third one was a nonspecific binding. The binding of N-deethylated dorzolamide to human CA-I and CA-II was competitively inhibited by dorzolamide, indicating that these compounds bind to the same binding site on CAs. In each other's presence, most of dorzolamide in erythrocytes binds to CA-II, whereas the N-deethylated metabolite mainly binds to CA-I.(ABSTRACT TRUNCATED AT 250 WORDS)


Dissociable Effects of Serotonin and Dopamine on the Valuation of Harm in Moral Decision Making.
An aversion to harming others is a core component of human morality and is disturbed in antisocial behavior. Deficient harm aversion may underlie instrumental and reactive aggression, which both feature in psychopathy. Past work has highlighted monoaminergic influences on aggression, but a mechanistic account of how monoamines regulate antisocial motives remains elusive. We previously observed that most people show a greater aversion to inflicting pain on others than themselves. Here, we investigated whether this hyperaltruistic disposition is susceptible to monoaminergic control. We observed dissociable effects of the serotonin reuptake inhibitor citalopram and the dopamine precursor levodopa on decisions to inflict pain on oneself and others for financial gain. Computational models of choice behavior showed that citalopram increased harm aversion for both self and others, while levodopa reduced hyperaltruism. The effects of citalopram were stronger than those of levodopa. Crucially, neither drug influenced the physical perception of pain or other components of choice such as motor impulsivity or loss aversion, suggesting a direct and specific influence of serotonin and dopamine on the valuation of harm. We also found evidence for dose dependency of these effects. Finally, the drugs had dissociable effects on response times, with citalopram enhancing behavioral inhibition and levodopa reducing slowing related to being responsible for another's fate. These distinct roles of serotonin and dopamine in modulating moral behavior have implications for potential treatments of social dysfunction that is a common feature as well as a risk factor for many psychiatric disorders.


Effect of metabolic acidosis on proximal tubular total CO2 absorption.
In vivo microperfusion studies of the proximal convoluted tubule of the rat were performed to determine the effect of metabolic acidosis on total CO2 (tCO2) absorption. In series I, tubular perfusion was performed in control and acidotic rats in a manner by which similar mean total CO2 concentrations in the proximal tubule were maintained. Comparable ranges of perfusion rate were studied in both groups. Following 3 days of HCl ingestion, plasma tCO2 was 20.0 +/- 0.9 mM in the acidotic rats whereas it was 29.6 +/- 0.53 mM in control rats. The arterial blood pH values were 7.25 +/- 0.02 vs. 7.43 +/- 0.01. Starting tCO2 perfusate concentrations were identical in both groups, 29.3 and 29.7 mM, as were the concentrations at the end of the perfused segments, 21.2 and 21.9 mM. The absorption of tCO2 (JtCO2, pmol X mm-1 X min-1) was significantly greater in the acidotic rats than in the controls, 576 +/- 39 vs. 256 +/- 21. At all perfusion rates studied, proximal tubular JtCO2 was higher in the acidotic than in the control rats. In series II, similar lengths of the late proximal tubule were perfused at the same rate in control and acidotic rats. Again, JtCO2 was higher in the acidotic rats, 352 +/- 19 vs. 198 +/- 13. The results indicate that at comparable luminal tCO2 concentration and tubular fluid flow rates, tCO2 absorption is significantly increased in the acidotic state. Although other mechanisms cannot be excluded, the finding of an increase in proximal tCO2 absorption in the acidotic rats is in agreement with the presence of an accelerated Na+/H+ exchange rate in brush border membrane vesicles obtained from the renal cortex of animals with metabolic acidosis.


Monitoring the hand hygiene compliance of health care workers in a general intensive care unit: Use of continuous closed circle television versus overt observation.
A variety of hand hygiene monitoring programs (HHMPs) have come into use in hospitals throughout the world. In the present study, we compare continuous closed circle television (CCTV) with overt observation for monitoring the hand hygiene compliance of health care workers (HCWs) in a general intensive care unit (GICU). This is a cross-sectional and comparative study. In this study, we use a novel hand hygiene CCTV monitoring system for hand hygiene performance monitoring. The study population incorporated all the GICU HCWs, including registered nurses, staff physicians, and auxiliary workers. All HCWs of our GICU were observed, including ICU registered nurses, ICU staff physicians, and auxiliary workers participated in the present study. Overall, each observer team did 50 sessions in each arm of the study. Total number of hand hygiene opportunities was approaching 500 opportunities. The compliance rates when only overt observations were performed was higher than when only covert observations were performed with a delta of approximately 10% (209 out of 590 [35.43%] vs 130 out of 533 [24.39%]; P < .001). Both methods of observations (overt and covert [CCTV]) demonstrated excellent reliability (intraclass correlation coefficient [ICC], 0.96 [0.93-0.98] of overt and ICC, 0.81 [0.69-0.89] for covert, respectively). However, the correlation between both methods was found weak in simultaneous sessions (ICC, 0.40 [0.62-0.107]). We demonstrated that CCTV is an appropriate, reliable, and neutral method for observation of hand hygiene. However, there is no clear basis for incorporating a CCTV observation modality into a health care system that already operates an overt observation program. We have shown that CCTV methodology records a different distribution of opportunities for performing hand hygiene and of actual performances of hand hygiene compared with overt observation.


Riding with Impaired Drivers Among Recent Latino Immigrants in Southern Florida.
In a previous effort we showed that compared with immigrants who are permanent residents, undocumented immigrants are more likely to binge drink, but less likely to drink while impaired (DWI) partly due to their limited amount of driving. This report examines a related risk: riding with an impaired driver (RWI). Data came from an ongoing longitudinal sample of Latino immigrants to Miami-Dade County, FL. Descriptive analyses and regression techniques were applied. While DWI rates among Latino immigrants is heavily limited by their access to a car, RWI rates were not restricted by driving limitations, nor related to participants' legal immigration status (LIS). RWI rates were linked only to heavy drinking. Because it is not affected by driving limitations, RWI for these Latino immigrants is perhaps a more immediate risk than DWI. Addressing RWI among Latino immigrants should be a priority for traffic safety to Miami/Dade country.


Extracting a maximum of useful information from statistical research data.
The practice of statistical inference in psychological research is critically reviewed. Particular emphasis is put on the fast pace of change from the sole reliance on null hypothesis significance testing (NHST) to the inclusion of effect size estimates, confidence intervals, and an interest in the Bayesian approach. We conclude that these developments are helpful for psychologists seeking to extract a maximum of useful information from statistical research data, and that seven decades of criticism against NHST is finally having an effect.


Matrix metalloproteinases cleave at two distinct sites on human cartilage link protein.
The actions of human recombinant stromelysins-1 and -2, collagenase, gelatinases A and B and matrilysin on neonatal human proteoglycan aggregates were examined. With the exception of gelatinase B, aggrecan was degraded extensively by most metalloproteinases studied, whereas link protein showed only limited proteolysis. Sequencing studies of modified link protein components revealed that stromelysins-1 and -2, gelatinases A and B and collagenase cleaved specifically between His16 and Ile17, and matrilysin, stromelysin-2 and gelatinase A cleaved between Leu25 and Leu26. Cleavage at the former bond generated a link protein component with the same N-terminus as that isolated from newborn human cartilage. Based on previously determined in situ cleavage sites it is evident that matrix metalloproteinases are not solely responsible for the accumulation of link protein degradation products in adult human cartilage, indicating that additional proteolytic agents are involved in the normal catabolism of human cartilage matrix.


Correlation of Female Indices From Virulence Assays on Inbred Lines and Field Populations of Heterodera glycines.
A lack of diversity and durability of resistant soybean varieties complicates management of the soybean cyst nematode (SCN), Heterodera glycines, exemplified by the current overdependence on the PI 88788 source of resistance. Of interest is the effect of adaptation of a SCN population to a source of resistance on its subsequent ability to develop on others. Female indices (FI) from virulence assays (race, HG Type and SCN Type tests) for SCN field populations and inbred lines were analyzed. Female indices on PI 88788, PI 209332 and PI 548316 were highly correlated, as were those of PI 548402, PI 90763, PI 89772 and PI 438489B. Previous studies on resistant SCN-infected soybean roots indicated that the cellular resistance response was similar within these two groups of soybean genotypes. In field populations, highly significant correlations were also found between FI on PI 88788 and PI 548402 and those on PI 89772 and PI 437654. In inbred lines, FI on PI 437654 were correlated with PI 90763 and PI 438489B. To avoid further adaptation, rotation of cultivars with resistance from these groups should be carefully monitored, including those from the most promising source of resistance, PI 437654, such as CystX. In a separate test, 10 soybean varieties developed from CystX were tested against HG Type 0, HG Type 2.5.7 and HG Type 1-7. Female development occurred in all tests but one. Although identification and deployment of unique resistance is needed, management strategies to prevent and detect adaptation should be emphasized.


[Evaluation of the incidence of dental caries in patients with Down syndrome after their insertion in a preventive program].
The objective of this work was to verify the incidence of dental caries by means of the CPO-D, CPO-S, ceo-d and ceo-s indexes in patients with Down syndrome regularly enrolled in a preventive program. Twenty four Down syndrome patients of both sexes age range of one to 48 years were examined. The prevalence (initial experimental situation) and incidence (final experimental situation) of dental caries were verified using of the initial and final CPO-D, CPO-S, ceo-d and ceo-s indexes of the participants. From 24 individual examined, 10 (42.0%) were free of caries. The prevalence of dental caries showed values of CPO-D= 2.33; CPO-S= 3.60; ceo-d= 1.75 e ceo-s= 2.80; while the incidence of caries showed values of 2.33; 3.80; 1.10 e 1.90, respectively. Down syndrome individuals evaluated in this study presented low level of caries and small incidence of new lesions, emphasizing the importance of the maintenance of these patients at preventive programs.


Adjuvant nivolumab versus ipilimumab (CheckMate 238 trial): Reassessment of 4-year efficacy outcomes in patients with stage III melanoma per AJCC-8 staging criteria.
Nivolumab was approved as adjuvant therapy for melanoma based on data from CheckMate 238, which enrolled patients per American Joint Committee on Cancer version 7 (AJCC-7) criteria. Here, we analyse long-term outcomes per AJCC-8 staging criteria compared with AJCC-7 results to inform clinical decisions for patients diagnosed per AJCC-8. In a double-blind, phase 3 trial (NCT02388906), patients aged ≥15 years with resected, histologically confirmed AJCC-7 stage IIIB, IIIC, or IV melanoma were randomised to receive nivolumab 3 mg/kg every 2 weeks or ipilimumab 10 mg/kg every 3 weeks for 4 doses and then every 12 weeks, both intravenously ≤1 year. Recurrence-free survival (RFS) and distant metastasis-free survival (DMFS) were assessed in patients with stage III disease, per AJCC-7 and AJCC-8. Per AJCC-7 staging, 42.4% and 57.3% of patients were in substage IIIB and IIIC, respectively; per AJCC-8, 1.1%, 30.4%, 62.8%, and 5.0% were in IIIA, IIIB, IIIC, and IIID. After 4 years' minimum follow-up, the AJCC-7 superior efficacy of nivolumab over ipilimumab in patients with resected stage III melanoma was preserved per AJCC-8 analysis. No statistically significant difference in RFS between stage III substage hazard ratios was observed per AJCC-7 or -8 staging criteria (interaction test: AJCC-7, P = 0.8115; AJCC-8, P = 0.1051; P = 0.8392 ((AJCC-7) and P = 0.8678 (AJCC-8) for DMFS). CheckMate 238 4-year RFS and DMFS outcomes are consistent per AJCC-7 and AJCC-8 staging criteria. Outcome benefits can therefore be translated for patients diagnosed per AJCC-8.


Clinical immunology review series: an approach to desensitization.
Allergen immunotherapy describes the treatment of allergic disease through administration of gradually increasing doses of allergen. This form of immune tolerance induction is now safer, more reliably efficacious and better understood than when it was first formally described in 1911. In this paper the authors aim to summarize the current state of the art in immunotherapy in the treatment of inhalant, venom and drug allergies, with specific reference to its practice in the United Kingdom. A practical approach has been taken, with reference to current evidence and guidelines, including illustrative protocols and vaccine schedules. A number of novel approaches and techniques are likely to change considerably the way in which we select and treat allergy patients in the coming decade, and these advances are previewed.


The impact of genetic diversity on gene essentiality within the Escherichia coli species.
Bacteria from the same species can differ widely in their gene content. In Escherichia coli, the set of genes shared by all strains, known as the core genome, represents about half the number of genes present in any strain. Although recent advances in bacterial genomics have unravelled genes required for fitness in various experimental conditions, most studies have focused on single model strains. As a result, the impact of the species' genetic diversity on core processes of the bacterial cell remains largely under-investigated. Here, we have developed a CRISPR interference platform for high-throughput gene repression that is compatible with most E. coli isolates and closely related species. We have applied it to assess the importance of ~3,400 nearly ubiquitous genes in three growth conditions in 18 representative E. coli strains spanning most common phylogroups and lifestyles of the species. Our screens revealed extensive variations in gene essentiality between strains and conditions. Investigation of the genetic determinants for these variations highlighted the importance of epistatic interactions with mobile genetic elements. In particular, we have shown how prophage-encoded defence systems against phage infection can trigger the essentiality of persistent genes that are usually non-essential. This study provides broad insights into the evolvability of gene essentiality and argues for the importance of studying various isolates from the same species under diverse conditions.


Research priorities for high-quality geriatric emergency care: medication management, screening, and prevention and functional assessment.
Geriatric adults represent an increasing proportion of emergency department (ED) users and can be particularly vulnerable to acute illnesses. Health care providers have recently begun to focus on the development of quality indicators (QIs) to define a minimal standard of care. The original objective of this project was to develop additional ED-specific QIs for older patients within the domains of medication management, screening and prevention, and functional assessment, but the quantity and quality of evidence were insufficient to justify unequivocal minimal standards of care for these three domains. Accordingly, the authors modified the project objectives to identify key research opportunities within these three domains that can be used to develop QIs in the future. Each domain was assigned one or two content experts who created potential QIs based on a systematic review of the literature, supplemented by expert opinion. Candidate QIs were then reviewed by four groups: the Society for Academic Emergency Medicine (SAEM) Geriatric Task Force, the SAEM Geriatric Interest Group, and audiences at the 2008 SAEM Annual Meeting and the 2009 American Geriatrics Society Annual Meeting, using anonymous audience response system technology as well as verbal and written feedback. High-quality evidence based on patient-oriented outcomes was insufficient or nonexistent for all three domains. The participatory audiences did not reach a consensus on any of the proposed QIs. Key research questions for medication management (three), screening and prevention (two), and functional assessment (three) are presented based on proposed QIs that the majority of participants accepted. In assessing a minimal standard of care by which to systematically derive geriatric QIs for medication management, screening and prevention, and functional assessment, compelling clinical research evidence is lacking. Patient-oriented research questions that are essential to justify and characterize future QIs within these domains are described.


Rationalizing blood transfusion in cardiac surgery: the impact of a red cell volume-based guideline on blood usage and clinical outcome.
Cardiac surgery is currently considered one of the heaviest users of red blood cells. An explanation may be found, in part, in considering the effect of the heavy clear fluid load associated with cardiopulmonary bypass. This may result in the artificial depression of haemoglobin concentration, overestimating the requirement for red cell transfusion if this is the sole parameter considered. To address this issue, we examined the impact of a red cell volume-based transfusion guideline on transfusion requirement. This was a single-centre, randomized controlled trial. The cohort of 86 patients was allocated to receive red cells as per the red cell volume guideline (group RCV) or standard haemoglobin concentration-based departmental policy (group C). Outcome measures were red cell transfusion and clinical outcome. All preoperative data were comparable between the two groups. A significantly fewer percentage of patients in group RCV were transfused red cells (RCV = 32.6% vs. C = 53.5%, P = 0.05). No significant difference was found between any of the outcome measures with the exception of median hospital stay (RCV = 5.9 days vs. C = 6.8 days, P = 0.02). In elective cardiac surgery patients, considering haemoglobin concentration alone may overestimate the requirement for red cell transfusion. More research is required to determine the impact of restrictive transfusion policies on clinical outcome following cardiac surgery.


Integrating trait-based empirical and modeling research to improve ecological restoration.
A global ecological restoration agenda has led to ambitious programs in environmental policy to mitigate declines in biodiversity and ecosystem services. Current restoration programs can incompletely return desired ecosystem service levels, while resilience of restored ecosystems to future threats is unknown. It is therefore essential to advance understanding and better utilize knowledge from ecological literature in restoration approaches. We identified an incomplete linkage between global change ecology, ecosystem function research, and restoration ecology. This gap impedes a full understanding of the interactive effects of changing environmental factors on the long-term provision of ecosystem functions and a quantification of trade-offs and synergies among multiple services. Approaches that account for the effects of multiple changing factors on the composition of plant traits and their direct and indirect impact on the provision of ecosystem functions and services can close this gap. However, studies on this multilayered relationship are currently missing. We therefore propose an integrated restoration agenda complementing trait-based empirical studies with simulation modeling. We introduce an ongoing case study to demonstrate how this framework could allow systematic assessment of the impacts of interacting environmental factors on long-term service provisioning. Our proposed agenda will benefit restoration programs by suggesting plant species compositions with specific traits that maximize the supply of multiple ecosystem services in the long term. Once the suggested compositions have been implemented in actual restoration projects, these assemblages should be monitored to assess whether they are resilient as well as to improve model parameterization. Additionally, the integration of empirical and simulation modeling research can improve global outcomes by raising the awareness of which restoration goals can be achieved, due to the quantification of trade-offs and synergies among ecosystem services under a wide range of environmental conditions.


Explaining the Neuroevolution of Fighting Creatures Through Virtual fMRI.
While interest in artificial neural networks (ANNs) has been renewed by the ubiquitous use of deep learning to solve high-dimensional problems, we are still far from general artificial intelligence. In this article, we address the problem of emergent cognitive capabilities and, more crucially, of their detection, by relying on co-evolving creatures with mutable morphology and neural structure. The former is implemented via both static and mobile structures whose shapes are controlled by cubic splines. The latter uses ESHyperNEAT to discover not only appropriate combinations of connections and weights but also to extrapolate hidden neuron distribution. The creatures integrate low-level perceptions (touch/pain proprioceptors, retina-based vision, frequency-based hearing) to inform their actions. By discovering a functional mapping between individual neurons and specific stimuli, we extract a high-level module-based abstraction of a creature's brain. This drastically simplifies the discovery of relationships between naturally occurring events and their neural implementation. Applying this methodology to creatures resulting from solitary and tag-team co-evolution showed remarkable dynamics such as range-finding and structured communication. Such discovery was made possible by the abstraction provided by the modular ANN which allowed groups of neurons to be viewed as functionally enclosed entities.


Towards a universal vaccine for avian influenza: protective efficacy of modified Vaccinia virus Ankara and Adenovirus vaccines expressing conserved influenza antigens in chickens challenged with low pathogenic avian influenza virus.
Current vaccines targeting surface proteins can drive antigenic variation resulting either in the emergence of more highly pathogenic viruses or of antigenically distinct viruses that escape control by vaccination and thereby persist in the host population. Influenza vaccines typically target the highly mutable surface proteins and do not provide protection against heterologous challenge. Vaccines which induce immune responses against conserved influenza epitopes may confer protection against heterologous challenge. We report here the results of vaccination with recombinant modified Vaccinia virus Ankara (MVA) and Adenovirus (Ad) expressing a fusion construct of nucleoprotein and matrix protein (NP+M1). Prime and boost vaccination regimes were trialled in different ages of chicken and were found to be safe and immunogenic. Interferon-γ (IFN-γ) ELISpot was used to assess the cellular immune response post secondary vaccination. In ovo Ad prime followed by a 4 week post hatch MVA boost was identified as the most immunogenic regime in one outbred and two inbred lines of chicken. Following vaccination, one inbred line (C15I) was challenged with low pathogenic avian influenza (LPAI) H7N7 (A/Turkey/England/1977). Birds receiving a primary vaccination with Ad-NP+M1 and a secondary vaccination with MVA-NP+M1 exhibited reduced cloacal shedding as measured by plaque assay at 7 days post infection compared with birds vaccinated with recombinant viruses containing irrelevant antigen. This preliminary indication of efficacy demonstrates proof of concept in birds; induction of T cell responses in chickens by viral vectors containing internal influenza antigens may be a productive strategy for the development of vaccines to induce heterologous protection against influenza in poultry.


The clinical application of cancer immunotherapy based on naturally circulating dendritic cells.
Dendritic cells (DCs) can initiate and direct adaptive immune responses. This ability is exploitable in DC vaccination strategies, in which DCs are educated ex vivo to present tumor antigens and are administered into the patient with the aim to induce a tumor-specific immune response. DC vaccination remains a promising approach with the potential to further improve cancer immunotherapy with little or no evidence of treatment-limiting toxicity. However, evidence for objective clinical antitumor activity of DC vaccination is currently limited, hampering the clinical implementation. One possible explanation for this is that the most commonly used monocyte-derived DCs may not be the best source for DC-based immunotherapy. The novel approach to use naturally circulating DCs may be an attractive alternative. In contrast to monocyte-derived DCs, naturally circulating DCs are relatively scarce but do not require extensive culture periods. Thereby, their functional capabilities are preserved, the reproducibility of clinical applications is increased, and the cells are not dysfunctional before injection. In human blood, at least three DC subsets can be distinguished, plasmacytoid DCs, CD141+ and CD1c+ myeloid/conventional DCs, each with distinct functional characteristics. In completed clinical trials, either CD1c+ myeloid DCs or plasmacytoid DCs were administered and showed encouraging immunological and clinical outcomes. Currently, also the combination of CD1c+ myeloid and plasmacytoid DCs as well as the intratumoral use of CD1c+ myeloid DCs is under investigation in the clinic. Isolation and culture strategies for CD141+ myeloid DCs are being developed. Here, we summarize and discuss recent clinical developments and future prospects of natural DC-based immunotherapy.


Correction of bias in self-reported sitting time among office workers - a study based on compositional data analysis.
Objective Emerging evidence suggests that excessive sitting has negative health effects. However, this evidence largely relies on research using self-reported sitting time, which is known to be biased. To correct this bias, we aimed at developing a calibration model estimating "true" sitting from self-reported sitting. Methods Occupational sitting time was estimated by self-reports (the International Physical Activity Questionnaire) and objective measurements (thigh-worn accelerometer) among 99 Swedish office workers at a governmental agency, at baseline and 3 and 12 months afterwards. Following compositional data analysis procedures, both sitting estimates were transformed into isometric log-ratios (ILR). This effectively addresses that times spent in various activities are inherently dependent and can be presented as values of only 0-100%. Linear regression was used to develop a simple calibration model estimating objectively measured "true" sitting ILR (dependent variable) from self-reported sitting ILR (independent variable). Additional self-reported variables were then added to construct a full calibration model. Performance of the models was assessed by root-mean-square (RMS) differences between estimated and objectively measured values. Models developed on baseline data were validated using the follow-up datasets. Results Uncalibrated self-reported sitting ILR showed an RMS error of 0.767. Simple and full calibration models (incorporating body mass index, office type, and gender) reduced this error to 0.422 (55%) and 0.398 (52%), respectively. In the validations, model performance decreased to 57%/62% (simple models) and 57%/62% (full models) for the two follow-up data sets, respectively. Conclusions Calibration adjusting for errors in self-reported sitting led to substantially more correct estimates of "true" sitting than uncalibrated self-reports. Validation indicated that model performance would change somewhat in new datasets and that full models perform no better than simple models, but calibration remained effective.


Theory of noncontact point thermal sensing by fiber-optic radiometry.
This paper formulates a theory of noncontact point thermal sensing by fiber-optic radiometry. This theory covers the field of mid- and far-infrared fibers that are suitable for low-temperature radiometry. However, new problems arise in the infrared range, the emission of thermal radiation from the fiber itself due to infrared absorption introduces perturbations into the radiometry, and this must be taken into consideration. The model presented is based on three-dimensional optical geometry of bounded and tunneling skew rays and yields an analytical expression for the inclination and the skewness angle distribution of the guided power collected by the fiber from various layers of a thermal body. The effective field of view, the surface resolution, and the temperature resolution of fiber-optic radiometry are discussed. Thermal sensing by direct coupling is shown to have an advantage over the coupling of a focusing lens located behind the fiber tip. A formulation of fiber emissivity is presented that quantifies the suppression of radiometric perturbations in fiber-optic thermal sensing. Bulk and surface absorption in the fiber core and cladding absorption are all taken into consideration deriving emissivity. Combining the transmissivity and emissivity of the fiber, we propose a measurable criterion, a figure of merit, for fiber-optic radiometry.


Cell Cycle Inhibition To Treat Sleeping Sickness.
African trypanosomiasis is caused by infection with the protozoan parasite Trypanosoma brucei During infection, this pathogen divides rapidly to high density in the bloodstream of its mammalian host in a manner similar to that of leukemia. Like all eukaryotes, T. brucei has a cell cycle involving the de novo synthesis of DNA regulated by ribonucleotide reductase (RNR), which catalyzes the conversion of ribonucleotides into their deoxy form. As an essential enzyme for the cell cycle, RNR is a common target for cancer chemotherapy. We hypothesized that inhibition of RNR by genetic or pharmacological means would impair parasite growth in vitro and prolong the survival of infected animals. Our results demonstrate that RNR inhibition is highly effective in suppressing parasite growth both in vitro and in vivo These results support drug discovery efforts targeting the cell cycle, not only for African trypanosomiasis but possibly also for other infections by eukaryotic pathogens.IMPORTANCE The development of drugs to treat infections with eukaryotic pathogens is challenging because many key virulence factors have closely related homologues in humans. Drug toxicity greatly limits these development efforts. For pathogens that replicate at a high rate, especially in the blood, an alternative approach is to target the cell cycle directly, much as is done to treat some hematologic malignancies. The results presented here indicate that targeting the cell cycle via inhibition of ribonucleotide reductase is effective at killing trypanosomes and prolonging the survival of infected animals.


Estimation of body fluids with bioimpedance spectroscopy: state of the art methods and proposal of novel methods.
Determination of body fluids is a useful common practice in determination of disease mechanisms and treatments. Bioimpedance spectroscopy (BIS) methods are non-invasive, inexpensive and rapid alternatives to reference methods such as tracer dilution. However, they are indirect and their robustness and validity are unclear. In this article, state of the art methods are reviewed, their drawbacks identified and new methods are proposed. All methods were tested on a clinical database of patients receiving growth hormone replacement therapy. Results indicated that most BIS methods are similarly accurate (e.g. < 0.5 ± 3.0% mean percentage difference for total body water) for estimation of body fluids. A new model for calculation is proposed that performs equally well for all fluid compartments (total body water, extra- and intracellular water). It is suggested that the main source of error in extracellular water estimation is due to anisotropy, in total body water estimation to the uncertainty associated with intracellular resistivity and in determination of intracellular water a combination of both.


Epidemiology of community-onset Clostridium difficile infection in a community in the South of England.
Clostridium difficile infection (CDI) has traditionally been considered a hospital acquired infection but there are a rising number of infections in the community. This study estimates the prevalence of community-onset CDI (CO-CDI), defined as onset of symptoms in a community setting and outside the hospital, and examines the risk factors for CO-CDI in 2-64 year-olds. A standard questionnaire was used to retrospectively obtain information on the CDI risk factors of 58 cases of CO-CDI diagnosed between 1st April 2008 and 31st March 2009 in a community in the South of England. Each case was reviewed for the presence of 'established' risk factors for CDI, i.e., age ≥65 years, in-patient hospital stay, and recent (within ≤4 weeks) receipt of broad spectrum antibiotics, and other, 'non-established' risk factors for CDI, such as exposure to antibiotics more than 4 weeks preceding symptom onset, out-patient and day-surgery hospital exposure, contact with a hospitalised patient, and travel outside of the UK. Fifty-eight cases of CO-CDI were diagnosed among a total community population of 418,000, representing an estimated prevalence of CO-CDI of 1.29 per 10,000. All 58 cases were successfully contacted, representing a 100% response rate. Four cases were excluded from further analysis due to co-infection with Salmonella spp. and Campylobacter spp. Cases were more likely to be female, aged between 31 and 40 years, and present in the spring season (March-May), 2009. 46.3% (25/54) of cases had established risk factors for CDI, 20.4% (11/54) had non-established risk factors, 16.7% (9/54) had no risk factors and in the remaining 16.7% (9/54), available information was insufficient to classify by risk factor category. This study suggests that CDI should be included in the differential diagnosis of community-onset diarrhea in patients with or without established risk factors for CDI.


TDP-43 in familial and sporadic frontotemporal lobar degeneration with ubiquitin inclusions.
TAR DNA-binding protein 43 (TDP-43) is a major pathological protein of sporadic and familial frontotemporal lobar degeneration with ubiquitin-positive, tau-negative inclusions (FTLD-U) with or without motor neuron disease (MND). Thus, TDP-43 defines a novel class of neurodegenerative diseases called TDP-43 proteinopathies. We performed ubiquitin and TDP-43 immunohistochemistry on 193 cases of familial and sporadic FTLD with or without MND. On selected cases, immunoelectron microscopy and biochemistry were performed. Clinically defined frontotemporal dementias (FTDs) included four groups: 1) familial FTD with mutations in progranulin (n = 36), valosin-containing protein (n = 5), charged multivesicular body protein 2B (n = 4), and linked to chromosome 9p (n = 7); 2) familial cases of FTD with unknown gene association (n = 29); 3) sporadic FTD (n = 72); and 4) familial and sporadic FTD with MND (n = 40). Our studies confirm that the spectrum of TDP-43 proteinopathies includes most cases of sporadic and familial FTLD-U with and without MND and expand this disease spectrum to include reported families with FTD linked to chromosome 9p but not FTD with charged multivesicular body protein 2B mutations. Thus, despite significant clinical, genetic, and neuropathological heterogeneity of FTLD-U, TDP-43 is a common pathological substrate underlying a large subset of these disorders, thereby implicating TDP-43 in novel and unifying mechanisms of FTLD pathogenesis.