Improving Safe and Effective Use of Drugs in Pregnancy and Lactation: Workshop Summary.
In February 2015, given high rates of use of medications by pregnant women and the relative lack of data on safety and efficacy of many drugs utilized in pregnancy, the Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), the Society for Maternal-Fetal Medicine (SMFM), the American College of Obstetricians and Gynecologists (ACOG), and the American Academy of Pediatrics (AAP) convened a group of experts to review the "current" state of the clinical care and science regarding medication use during the perinatal period. The expert panel chose select medications to demonstrate what existing safety and efficacy data may be available for clinicians and patients when making decisions about use in pregnancy or lactation. Furthermore, these example medications also provided opportunities to highlight where data are lacking, thus forming a list of research gaps. Last, after reviewing the existing vaccine safety surveillance system as well as the legislative history surrounding the use of drugs for pediatric diseases, the expert panel made specific recommendations concerning policy efforts to stimulate more research and regulatory attention on drugs for pregnant and lactating women.


Clinically defined vascular depression.
The authors' goal was to examine the clinical presentation of a group of depressed elderly patients with clinically defined risk factors for vascular depression compared with a group of elderly depressed patients without such risk factors. Cognitive deficits, disability, and depressive symptoms were examined in 33 consecutively recruited elderly patients defined as having vascular depression and 32 patients defined as having nonvascular depression according to their scores on the Cumulative Illness Rating Scale-Geriatrics. The patients with vascular depression had greater overall cognitive impairment and disability than those with nonvascular depression. Fluency and naming were more impaired in patients with vascular depression, and they had more retardation and less agitation as well as less guilt feelings and greater lack of insight. The symptoms of vascular depression are consistent with lesions that may damage striato-pallido-thalamo-cortical pathways and other areas. The concept of vascular depression can provide the impetus for investigations of prevention and treatment of cerebrovascular disease and for studies of the course of vascular depression and selection of antidepressants.


The Initial Detection and Partial Characterization of Circulating Tumor Cells in Neuroendocrine Prostate Cancer.
The transition of prostate adenocarcinoma to a predominantly androgen receptor (AR) signaling independent phenotype can occur in the later stages of the disease and is associated with low AR expression +/- the development of small-cell or neuroendocrine tumor characteristics. As metastatic tumor biopsies are not always feasible and are difficult to repeat, we sought to evaluate noninvasive methods to identify patients transitioning toward a neuroendocrine phenotype (NEPC). We prospectively studied a metastatic tumor biopsy, serum biomarkers, and circulating tumor cells (CTC, Epic Sciences) from patients with castration-resistant prostate cancer (CRPC) including those with pure or mixed NEPC histology present on biopsy. CTCs labeled with the patient's clinical status were used to learn features that discriminate NEPC patients, which was then applied to an independent cohort. Twenty-seven patients with CRPC including 12 NEPC and 5 with atypical clinical features suggestive of NEPC transition were studied. CTCs from NEPC patients demonstrated frequent clusters, low or absent AR expression, lower cytokeratin expression, and smaller morphology relative to typical CRPC. A multivariate analysis of protein and morphologic variables enabled distinguishing CTCs of NEPC from CRPC. This CTC classifier was applied to an independent prospective cohort of 159 metastatic CRPC patients and identified in 17/159 (10.7%) of cases, enriched in patients with high CTC burden (P < 0.01) and visceral metastases (P = 0.04). CTCs from patients with NEPC have unique morphologic characteristics, which were also identified in a subset of CRPC patients with aggressive clinical features potentially undergoing NEPC transition.


A familial natural short sleep mutation promotes healthy aging and extends lifespan in Drosophila.
Sleep loss typically imposes negative effects on animal health. However, humans with a rare genetic mutation in the dec2 gene ( dec2 P384R ) present an exception; these individuals sleep less without the usual effects associated with sleep deprivation. Thus, it has been suggested that the dec2 P384R mutation activates compensatory mechanisms that allows these individuals to thrive with less sleep. To test this directly, we used a Drosophila model to study the effects of the dec2 P384R mutation on animal health. Expression of human dec2 P384R in fly sleep neurons was sufficient to mimic the short sleep phenotype and, remarkably, dec2 P384R mutants lived significantly longer with improved health despite sleeping less. The improved physiological effects were enabled, in part, by enhanced mitochondrial fitness and upregulation of multiple stress response pathways. Moreover, we provide evidence that upregulation of pro-health pathways also contributes to the short sleep phenotype, and this phenomenon may extend to other pro-longevity models.


Does amplitude-integrated electroencephalogram background pattern correlate with cerebral injury in neonates with hypoxic-ischaemic encephalopathy?
To determine the correlation between amplitude-integrated electroencephalogram (aEEG) background pattern and cerebral magnetic resonance imaging (MRI) in infants with hypoxic-ischaemic encephalopathy (HIE) and to examine whether the correlation changes with therapeutic hypothermia. We included 38 term-born infants with HIE of whom 17 were cooled. All were continuously monitored with aEEG. Background pattern was scored at the beginning and the end of the recording. Cerebral MRI was obtained on median day 5 (2-11 days). Abnormalities were classified using a predefined scoring system for basal ganglia, watershed and overall injury, and then grouped into mild-moderate and severe. Abnormal aEEG background pattern correlated with more severe cerebral injury on MRI in the non-cooled infants (P < 0.01). In addition, cooled infants had less severe cerebral injury than non-cooled infants, in particular on T2-weighted images (watershed P = 0.04 and total injury score = 0.07). Abnormal aEEG background pattern is predictive of abnormal MRI, but therapeutic hypothermia seems to reduce this association. Thus, when cooling is applied in a clinical setting, the predictive value of aEEG may be limited.


Protective Effects of Vitamin E Consumption against 3MT Electromagnetic Field Effects on Oxidative Parameters in Substantia Nigra in Rats.
Electromagnetic fields (EMFs) can influence the biological system by the formation of free radicals in cells. The EMFs are able to deteriorate defense system against free radicals that leads to oxidative stress (OS). Lipid peroxidation process (LPO) is an index of oxidative stress, and the Malandialdehyde (MDA) is the final product of LPO. Vitamin E is the most important antioxidant which inhibits the LPO process. The aim of this study was to evaluate the effects of 3MT EMF exposure on oxidative stress parameters in substantia nigra and the role of vitamin E in reducing oxidative stress and preventing of LPO process. 40 male Wistar rats were randomly divided into 4 groups: 1) Control group: received standard food without exposure to EMF and without consumption of vitamin E, 2) Experimental group 1: was exposed to EMF (3MT) 4 h/day for 50 days, 3) The experimental group 2: received 200 mg/kg vitamin E with gavage every day and also was exposed to EMF (3MT) 4 h/day for 50 days, 4) Sham group: received water with gavage for 50 days. A significant increase in MDA levels and Glutation peroxidase (GSH-Px) activity of the substantia nigra following 50 days exposure to EMF was detected, but the superoxide dismutase (SOD) activity was decreased. Exposure did not change total antioxidant capacity (TAC) levels in plasma. Vitamin E treatment significantly prevented the increase of the MDA levels and GSHPx activity and also prevented the decrease of SOD activity in tissue but did not alter TAC levels. The GSH-Px activity increased because the duration and intensity of exposure were not enough to decrease it. We demonstrated two important findings; that 50 days exposure to 3 MT electromagnetic field caused oxidative stress by increasing the levels of MDA, and decreasing SOD activity in the substantia nigra; and that treatment with the vitamin E significantly prevented the oxidative stress and lipid peroxidation.


Whole-body calorimetry studies in adult men. 1. The effect of fat over-feeding on 24 h energy expenditure.
Eight young men of normal weight were maintained for 1 week on a weight-maintenance diet followed by a 1-week period of over-feeding with extra fat designed to increase energy intake by 50%. Two 36 h calorimetry sessions with low and high physical activities were included in each feeding period. Faecal and urine collections permitted checks on energy malabsorption and nitrogen excretion. Over-feeding led to increases in body-weight, faecal energy and N excretion and in estimated N retention. Faecal energy outputs on the maintenance and over-feeding diets were 5 and 4.4% of the respective gross energy intakes. Energy expenditure on fat over-feeding increased by 5.6% on the low-activity regimen and 6.4% on the high-activity regimen. This amounted, in terms of the extra energy intake, to 9 and 11% on the inactive and active schedules respectively. The increase affected day- and night-time rates of energy expenditure plus the basal metabolic rate. Individuals with a low percentage body fat showed the greatest response to over-feeding. Nutrient-balance studies derived from calorimetry suggested that fat over-feeding led to substantial fat deposition with no evidence of sparing of carbohydrate oxidation. The theoretical cost of depositing dietary fat was exceeded, suggesting that regulatory thermogenic mechanisms may have been stimulated to a small extent.


Mechanisms of drug resistance in cancer chemotherapy: coordinated role and regulation of efflux transporters and metabolizing enzymes.
Cancer remains one of the major leading causes of death worldwide. Acquisition of multidrug resistance (MDR) remains a major impediment to successful chemotherapy. As the name implies, MDR is not limited only to one drug but often associated to structurally and functionally unrelated chemotherapeutics. Extensive research and investigations have identified several mechanisms underlying the development of MDR. This process of drug resistance is considered to be multifactorial including decreased drug accumulation, increased efflux, increased biotransformation, drug compartmentalization, modification of drug targets and defects in cellular pathways. In the first part of the review, these pharmacokinetic and pharmacodynamic mechanisms have been described in brief. Although the pathways can act independently, they are more often intertwined. Of the various mechanisms involved, up-regulation of efflux transporters and metabolizing enzymes constitute a major resistance phenotype. This review also provides a general biological overview of important efflux transporters and metabolizing enzymes involved in MDR. Further, synergistic action between efflux transporters and metabolizing enzymes leading to MDR could possibly arise due to two different factors; overlapping substrate specificity and coordinated regulation of their expression. The expression of efflux transporters and metabolizing enzymes is governed by nuclear receptors, mainly pregnane X receptor (PXR). The pharmacological role of PXR and advances in the development of PXR antagonists to overcome MDR are outlined.


Distinct expression and function of the novel mouse chemokine monocyte chemotactic protein-5 in lung allergic inflammation.
We have cloned a novel mouse CC chemokine cDNA from the lung during an allergic inflammatory reaction. The protein encoded by this cDNA is chemotactic for eosinophils, monocytes, and lymphocytes in vitro and in vivo. Based on its similarities in sequence and function with other CC chemokines, we have named it mouse monocyte chemotactic protein-5 (mMCP-5). Under noninflammatory conditions, expression of mMCP-5 in the lymph nodes and thymus is constitutive and is generally restricted to stromal cells. Neutralization of mMCP-5 protein with specific antibodies during an allergic inflammatory reaction in vivo resulted in a reduction in the number of eosinophils that accumulated in the lung. Moreover, mMCP-5 mRNA expression in vivo is regulated differently from that of other major CC chemokines in the lung during the allergic reaction, including Eotaxin. The presence of lymphocytes is essential for expression of mMCP-5 by alveolar macrophages and smooth muscle cells in the lung, and the induction of mMCP-5 RNA occurs earlier than that of the eosinophil chemokine Eotaxin during allergic inflammation. In contrast to Eotaxin, mRNA for mMCP-5 can be produced by mast cells. From these results, we postulate that mMCP-5 plays a pivotal role during the early stages of allergic lung inflammation.


Effect of enrofloxacin on Haemophilus parasuis infection, disease and immune response.
Haemophilus parasuis, the causative agent of Glasser's disease, is a pathogen that colonizes the upper respiratory tract (URT) of pigs, invades the bloodstream and causes polyserositis. Because antimicrobials are highly effective against H. parasuis, we hypothesized that they could have a detrimental effect on the establishment of an immune response if given at the time of URT colonization. In this study, we characterized clinical outcomes and antibody and IFN-γ responses to H. parasuis in pigs treated with enrofloxacin before or after low dose inoculation with a pathogenic H. parasuis strain. Pigs that were only inoculated with the agent (EXP group) and pigs that were treated with enrofloxacin and then inoculated (ABT/EXP group) developed signs of disease starting at 4days post inoculation (DPI), presented a significant increase in serum IgG and were protected against a subsequent homologous challenge. In contrast, pigs treated with antibiotic after inoculation (EXP/ABT group) neither showed signs of disease nor seroconverted (IgG) after low dose inoculation. EXP/ABT pigs as well as naïve control pigs [enrofloxacin only (ABT) and challenge only (CHA)] were susceptible to challenge. Variable levels of antibodies in bronchioalveolar fluid and IFN-γ in peripheral blood mononuclear cells were observed after H. parasuis inoculation, but were not associated with protection. In summary, only pigs treated before low dose H. parasuis inoculation seroconverted and were protected against subsequent challenge. Results from this study can help determine timing of antimicrobial use and contribute to our current understanding of judicious antibiotic use.


Cold-adapted poliovirus mutants bypass a postentry replication block.
In the current model of poliovirus entry, the initial interaction of the native virion with its cellular receptor is followed by a transition to an altered form, which then acts as an intermediate in viral entry. While the native virion sediments at 160S in a sucrose gradient, the altered particle sediments at 135S, has lost the coat protein VP4, and has become more hydrophobic. Altered particles can be found both associated with cells and in the culture medium. It has been hypothesized that the cell-associated 135S particle releases the viral genome into the cell cytoplasm and that nonproductive transitions to the 135S form are responsible for the high particle-to-PFU ratio observed for polioviruses. At 25 degrees C, a temperature at which the transition to 135S particles does not occur, the P1/Mahoney strain of poliovirus was unable to replicate, and cold-adapted (ca) mutants were selected from the population. These mutants have not gained the ability to convert to 135S particles at 25 degrees C, and the block to wild-type (wt) infection at low temperatures is not at the level of cellular entry. The particle-to-PFU ratio of poliovirus does not change at 25 degrees C in the absence of alteration. Three independent amino acid changes in the 2C coding region were identified in ca mutants, at positions 218 (Val to Ile), 241 (Arg to Ala), and 309 (Met to Val). Introduction of any of these mutations individually into wt poliovirus by site-directed mutagenesis confers the ca phenotype. All three serotypes of the Sabin vaccine strains and the P3/Leon strain of poliovirus also exhibit the ca phenotype. These results do not support a model of poliovirus entry into cells that includes an obligatory transition to the 135S particle.


Protean and boundaryless career orientations: A critical review and meta-analysis.
The protean/boundaryless career concepts refer to people becoming more self-directed and flexible in managing their careers in response to societal shifts in work arrangements. A sizable literature has emerged on protean/boundaryless career orientations/preferences (PBCO). Questions remain, however, about the structure of PBCO and whether they predict important criteria. The PBCO literature is largely disconnected from broader individual-level career research, making it unclear how PBCO intersect with career models based on other characteristics. We address these questions by systematically reviewing/meta-analyzing PBCO research. On the basis of 135 demographically/occupationally diverse samples from 35 countries (45,288 individuals), we find no support for traditional distinctions between protean and boundaryless orientations-protean self-directed, protean values-driven, and boundaryless psychological mobility all load onto a single general factor, labeled proactive career orientation, and are only weakly related to boundaryless physical mobility preferences. We also find that PBCO predict career self-management behaviors and career satisfaction but are less related to non-career-focused attitudes, objective success, or physical mobility behavior. PBCO are strongly related to proactivity-related and self-efficacy personality traits. We use these findings to propose an integrative model for how PBCO and other dispositions mutually influence career behavior. We discuss when PBCO may have advantages over broad traits for understanding careers, implications for counseling practice, and directions for future research. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


The role of the endocannabinoid system in aetiopathogenesis of endometriosis: A potential therapeutic target.
Endometriosis affects a large proportion of women during their reproductive years and is associated with pain and infertility, also affecting psychological wellbeing and quality of life. The pathogenesis of the disease remains unclear, although it is believed to be multifactorial. The endocannabinoid system (ECS) consists of a number of ligands, receptors and enzymes, and has gained interests in endometriosis research. This review aims to summarise all available evidence reporting the roles of the ECS in endometriosis. A literature search of the PubMed, EMBASE, and Web of Science electronic medical databases was performed. Original and review articles published in peer-reviewed journals were included. No publication date or publication status restrictions were imposed. Significant differences in the concentrations and expressions of the components of the ECS were reported in the eutopic and ectopic endometrium, and the systemic circulation of women with endometriosis compared to controls. Endometriosis appears to be associated with downregulation of CB1 receptors and upregulation of TRPV1 receptors. The role of CB1 and progesterone in anti-inflammatory action and the role of TRPV1 in inflammation and pain are of particular interests. Furthermore, the ECS has been reported to be involved in processes relevant to endometriosis, including cell migration, cell proliferation, apoptosis, inflammation, and interacts with sex steroid hormones. The ECS may play a role in disease establishment, progression, and pain in endometriosis. However, reports are based on studies of limited size and there are inconsistencies among the definition of their control groups. There are also conflicting reports regarding precise involvement of the ECS in endometriosis. Future research with larger numbers, strict inclusion and exclusion criteria and detailed clinical information is imperative.


The RING-H2-finger protein APC11 as a target of hydrogen peroxide.
The anaphase-promoting complex (APC) is a ubiquitin-protein ligase (E3) that targets cell cycle regulators such as cyclin B and securin for degradation. The APC11 subunit functions as the catalytic core of this complex and mediates the transfer of ubiquitin from a ubiquitin-conjugating enzyme (E2) to the substrate. APC11 contains a RING-H2-finger domain, which includes one histidine and seven cysteine residues that coordinate two Zn(2+) ions. We now show that exposure of purified APC11 to H(2)O(2) (0.1 to 1 mM) induced the release of bound zinc as a result of the oxidation of cysteine residues. It also impaired the physical interaction between APC11 and the E2 enzyme Ubc4 as well as inhibited the ubiquitination of cyclin B1 by APC11. The release of HeLa cells from metaphase arrest in the presence of exogenous H(2)O(2) inhibited the ubiquitination of cyclin B1 as well as the degradation of cyclin B1 and securin that were apparent in the absence of H(2)O(2). The presence of H(2)O(2) also blocked the co-immunoprecipitation of Ubc4 with APC11 and delayed the exit of cells from mitosis. Inhibition of APC11 function by H(2)O(2) thus likely contributes to the delay in cell cycle progression through mitosis that is characteristic of cells subjected to oxidative stress.


A unique subset of low-risk Wilms tumors is characterized by loss of function of TRIM28 (KAP1), a gene critical in early renal development: A Children's Oncology Group study.
This study explores the genomic alterations that contribute to the formation of a unique subset of low-risk, epithelial differentiated, favorable histology Wilms tumors (WT), tumors that have been characterized by their expression of post-induction renal developmental genes (Subset 1 WT). We demonstrate copy neutral loss of heterozygosity involving 19q13.32-q13.43, unaccompanied by evidence for imprinting by DNA methylation. We further identified loss-of-function somatic mutations in TRIM28 (also known as KAP1), located at 19q13, in 8/9 Subset 1 tumors analyzed. An additional germline TRIM28 mutation was identified in one patient. Retrospective evaluation of previously analyzed WT outside of Subset 1 identified an additional tumor with anaplasia and both TRIM28 and TP53 mutations. A major function of TRIM28 is the repression of endogenous retroviruses early in development. We depleted TRIM28 in HEK293 cells, which resulted in increased expression of endogenous retroviruses, a finding also demonstrated in TRIM28-mutant WT. TRIM28 has been shown by others to be active during early renal development, and to interact with WTX, another gene recurrently mutated in WT. Our findings suggest that inactivation of TRIM28 early in renal development contributes to the formation of this unique subset of FHWTs, although the precise manner in which TRIM28 impacts both normal renal development and oncogenesis remains elusive.


Fabry-Pérot optical filter assembly: a candidate for the Mie/ Rayleigh separator in EarthCARE.
This paper describes the development, characterisation and environmental testing of a high performance state-of-the-art optical filter for separating the Mie and Rayleigh backscatter signals for the ATLID LIDAR of the EarthCARE mission. The filter assembly utilises capacitance stabilised Fabry-Pérot étalon technology to provide high resolution optical filtering of the LIDAR signal, efficiently separating the two critical backscatter components from the atmosphere. The paper describes the development of the filter assembly and its subsequent performance testing. The filter has demonstrated state-of-the-art optical performance. Additionally, one of the étalons has been taken through complete environmental testing for space applications.


Undoing a quantum measurement.
In general, a quantum measurement yields an undetermined answer and alters the system to be consistent with the measurement result. This process maps multiple initial states into a single state and thus cannot be reversed. This has important implications in quantum information processing, where errors can be interpreted as measurements. Therefore, it seems that it is impossible to correct errors in a quantum information processor, but protocols exist that are capable of eliminating them if they affect only part of the system. In this work we present the deterministic reversal of a fully projective measurement on a single particle, enabled by a quantum error-correction protocol in a trapped ion quantum information processor. We further introduce an in-sequence, single-species recooling procedure to counteract the motional heating of the ion string due to the measurement.


Regulation of the expression level of transcription factor XylS reveals new functional insight into its induction mechanism at the Pm promoter.
XylS is the positive regulator of the inducible Pm promoter, originating from Pseudomonas putida, where the system controls a biochemical pathway involved in degradation of aromatic hydrocarbons, which also act as inducers. The XylS/Pm positive regulator/promoter system is used for recombinant gene expression and the output from Pm is known to be sensitive to the intracellular XylS concentration. By constructing a synthetic operon consisting of xylS and luc, the gene encoding luciferase, relative XylS expression levels could be monitored indirectly at physiological concentrations. Expression of XylS from inducible promoters allowed control over a more than 800-fold range, however, the corresponding output from Pm covered only an about five-fold range. The maximum output from Pm could not be increased by introducing more copies of the promoter in the cells. Interestingly, a previously reported XylS variant (StEP-13), known to strongly stimulate expression from Pm, caused the same maximum activity from Pm as wild-type XylS at high XylS expression levels. Under uninduced conditions expression from Pm also increased as a function of XylS expression levels, and at very high concentrations the maximum activity from Pm was the same as in the presence of inducer. According to our proposed model, which is in agreement with current knowledge, the regulator, XylS, can exist in three states: monomers, dimers, and aggregates. Only the dimers are active and able to induce expression from Pm. Their maximum intracellular concentration and the corresponding output from Pm are limited by the concentration-dependent conversion into inactive aggregates. Maximization of the induction ratio at Pm can be obtained by expression of XylS at the level where aggregation occurs, which might be exploited for recombinant gene expression. The results described here also indicate that there might exist variants of XylS which can exist at higher active dimer concentrations and thus lead to increased expression levels from Pm.


ThermoData Engine (TDE): software implementation of the dynamic data evaluation concept. 8. Properties of material streams and solvent design.
ThermoData Engine (TDE) is the first full-scale software implementation of the dynamic data evaluation concept, as reported in this journal. The present paper describes the first application of this concept to the evaluation of thermophysical properties for material streams involving any number of chemical components with assessment of uncertainties. The method involves construction of Redlich-Kister type equations for individual properties (excess volume, thermal conductivity, viscosity, surface tension, and excess enthalpy) and activity-coefficient models for phase equilibrium properties (vapor-liquid equilibrium). Multicomponent models are based on those for the pure-components and all binary subsystems evaluated on demand through the TDE software algorithms. Models are described in detail, and extensions to the class structure of the program are provided. Novel program features, such as ready identification of key measurements for subsystems that can reduce the combined uncertainty for a particular stream property, are described. In addition, new product-design features are described for selection of solvents for optimized crystal dissolution, separation of binary crystal mixtures, and solute extraction from a single-component solvent. Planned future developments are summarized.


Effects of water-emission anisotropy on multispectral remote sensing at thermal wavelengths of ocean temperature and of cirrus clouds.
The assumption of blackbody emission (emissivity, 1.0) for a calm ocean surface can lead to significant underestimates of the sea-surface temperature (SST) derived from IR radiometric data. Taking the optical properties of the atmosphere as known, we calculate the errors stemming from the blackbody assumption for cases of a purely absorbing or a purely scattering atmosphere. It is observed that for an absorbing atmosphere the errors in SST are always reduced and are the same whether measurements are made from space or at any level in the atmosphere. As for atmospheric scattering, the SST errors are slightly reduced when one is viewing from large zenith angles but are slightly enhanced when one is viewing from the zenith. The inferred optical thickness tau of an absorbing layer can be in error under the blackbody assumption by a Deltatau of 0.01-0.08, while the inferred optical thickness of a scattering layer can be in error by a larger amount, Deltatau of 0.03-0.13. The error Deltatau depends only weakly on the actual optical thickness and on the viewing angle, but it is rather sensitive to the wavelength of the measurement. In the absence of steep slopes in the wave-slope distribution, directional emissivities are essentially unchanged by sea state when one is viewing from or near the zenith. When one is viewing from moderately large zenith angles (such as 507 degrees ), however, the departures in the directional emissivities from blackbody emission can be much larger under perturbed sea state than under calm conditions.


Functional interactions between BRCA1 and the checkpoint kinase ATR during genotoxic stress.
The BRCA1 gene encodes a tumor suppressor that is mutated in 50% of familial breast cancers. The BRCA1 protein has been implicated in the DNA damage response, as DNA damage induces the phosphorylation of BRCA1 and causes its recruitment into nuclear foci that contain DNA repair proteins. The ataxia-telangiectasia-mutated (ATM) gene product controls overall BRCA1 phosphorylation in response to gamma-irradiation (IR). In this study, we show that BRCA1 phosphorylation is only partially ATM dependent in response to IR and ATM independent in response to treatment with UV light, or the DNA replication inhibitors hydroxyurea (HU) and aphidicolin (APH). We provide evidence that the kinase responsible for this phosphorylation is the ATM-related kinase, ATR. ATR phosphorylates BRCA1 on six Ser/Thr residues, including Ser 1423, in vitro. Increased expression of ATR enhanced the phosphorylation of BRCA1 on Ser 1423 following cellular exposure to HU or UV light, whereas doxycycline-induced expression of a kinase-inactive ATR mutant protein inhibited HU- or UV light-induced Ser 1423 phosphorylation in GM847 fibroblasts, and partially suppressed the phosphorylation of this site in response to IR. Thus, ATR, like ATM, controls BRCA1 phosphorylation in vivo. Although ATR isolated from DNA-damaged cells does not show enhanced kinase activity in vitro, we found that ATR responds to DNA damage and replication blocks by forming distinct nuclear foci at the sites of stalled replication forks. Furthermore, ATR nuclear foci overlap with the nuclear foci formed by BRCA1. The dramatic relocalization of ATR in response to DNA damage points to a possible mechanism for its ability to enhance the phosphorylation of substrates in response to DNA damage. Together, these results demonstrate that ATR and BRCA1 are components of the same genotoxic stress-responsive pathway, and that ATR directly phosphorylates BRCA1 in response to damaged DNA or stalled DNA replication.


Ape conservation physiology: fecal glucocorticoid responses in wild Pongo pygmaeus morio following human visitation.
Nature-based tourism can generate important revenue to support conservation of biodiversity. However, constant exposure to tourists and subsequent chronic activation of stress responses can produce pathological effects, including impaired cognition, growth, reproduction, and immunity in the same animals we are interested in protecting. Utilizing fecal samples (N = 53) from 2 wild habituated orangutans (Pongo pygmaeus morio) (in addition to 26 fecal samples from 4 wild unhabituated orangutans) in the Lower Kinabatangan Wildlife Sanctuary of Sabah, Malaysian Borneo, we predicted that i) fecal glucocorticoid metabolite concentrations would be elevated on the day after tourist visitation (indicative of normal stress response to exposure to tourists on the previous day) compared to samples taken before or during tourist visitation in wild, habituated orangutans, and ii) that samples collected from habituated animals would have lower fecal glucocorticoid metabolites than unhabituated animals not used for tourism. Among the habituated animals used for tourism, fecal glucocorticoid metabolite levels were significantly elevated in samples collected the day after tourist visitation (indicative of elevated cortisol production on the previous day during tourist visitation). Fecal glucocorticoid metabolite levels were also lower in the habituated animals compared to their age-matched unhabituated counterparts. We conclude that the habituated animals used for this singular ecotourism project are not chronically stressed, unlike other species/populations with documented permanent alterations in stress responses. Animal temperament, species, the presence of coping/escape mechanisms, social confounders, and variation in amount of tourism may explain differences among previous experiments. Acute alterations in glucocorticoid measures in wildlife exposed to tourism must be interpreted conservatively. While permanently altered stress responses can be detrimental, preliminary results in these wild habituated orangutans suggest that low levels of predictable disturbance can likely result in low physiological impact on these animals.


Discovery of the type VII ESX-1 secretion needle?
Mycobacterium tuberculosis, the etiological agent of human tuberculosis, harbours five ESAT-6/type VII secretion (ESX/T7S) systems. The first esx gene clusters were identified during the genome-sequencing project of M. tuberculosis H37Rv. Follow-up studies revealed additional genes playing important roles in ESX/T7S systems. Among the latter genes, one can find those that encode Pro-Glu (PE) and Pro-Pro-Glu (PPE) proteins as well as a gene cluster that is encoded >260 kb upstream of the esx-1 locus and encodes ESX-1 secretion-associated proteins EspA (Rv3616c), EspC (Rv3615c) and EspD (Rv3614c). The espACD cluster has been suggested to have an important function in ESX-1 secretion since EspA-EspC and EsxA-EsxB are mutually co-dependent on each other for secretion. However, the molecular mechanism of this co-dependence and interaction between the substrates remained unknown. In this issue of Molecular Microbiology, Lou and colleagues show that EspC forms high-molecular weight polymerization complexes that resemble selected components of type II, III and/or IV secretion systems of Gram-negative bacteria. Indeed, EspC-multimeric complexes form filamentous structures that could well represent a secretion needle of ESX-1 type VII secretion systems. This exciting observation opens new avenues for research to discover and characterize ESX/T7S components and elucidates the co-dependence of EsxA/B secretion with EspA/C.


MicroRNAome of porcine pre- and postnatal development.
The domestic pig is of enormous agricultural significance and valuable models for many human diseases. Information concerning the pig microRNAome (miRNAome) has been long overdue and elucidation of this information will permit an atlas of microRNA (miRNA) regulation functions and networks to be constructed. Here we performed a comprehensive search for porcine miRNAs on ten small RNA sequencing libraries prepared from a mixture of tissues obtained during the entire pig lifetime, from the fetal period through adulthood. The sequencing results were analyzed using mammalian miRNAs, the precursor hairpins (pre-miRNAs) and the first release of the high-coverage porcine genome assembly (Sscrofa9, April 2009) and the available expressed sequence tag (EST) sequences. Our results extend the repertoire of pig miRNAome to 867 pre-miRNAs (623 with genomic coordinates) encoding for 1,004 miRNAs, of which 777 are unique. We preformed real-time quantitative PCR (q-PCR) experiments for selected 30 miRNAs in 47 tissue-specific samples and found agreement between the sequencing and q-PCR data. This broad survey provides detailed information about multiple variants of mature sequences, precursors, chromosomal organization, development-specific expression, and conservation patterns. Our data mining produced a broad view of the pig miRNAome, consisting of miRNAs and isomiRs and a wealth of information of pig miRNA characteristics. These results are prelude to the advancement in pig biology as well the use of pigs as model organism for human biological and biomedical studies.


Subplasmalemmal calcium-binding microregions in Acanthamoeba.
Acanthamoeba cells, fixed with glutaraldehyde supplemented with calcium ions, show Ca-dependent, electron-dense deposits (CaDD) at the cytoplasmic surface of the plasma membrane. The formation of CaDD is stimulated by pre-incubation and fixation of the cells in the presence of ATP. Chemical analysis of the trichloroacetic acid extracts of CaDD-containing cells demonstrates that adenosine monophosphate, pyrophosphate and inorganic phosphate are probably the compounds involved in the formation of CaDD. Treatment of CaDD-containing cells with exogenous phosphatase evokes the disappearance of electron-dense material and a fine fibrillar network can be observed inside the 'empty' CaDD. The ability to restore CaDD in the presence of calcium ions with either pyrophosphate or orthophosphate confirms the suggestion that calcium/phosphate salts may be deposited in special subplasmalemmal calcium-binding microregions.


Synthesis and photophysical studies of an indigo derivative: N-octyl-7,7'-diazaindigo.
In this paper, we explore the synthesis, characterization, and photophysical properties of a novel indigo derivative, N-octyl-7,7'-diazaindigo, being the first time that diazaindigos have been studied as photophysically-active chemical entities. Reduction of the neutral "keto-form" to the so-called "leuco-form" changes the global spectroscopic and photophysical behaviors. Both species have been investigated by different photophysical studies, such as analysis of absorption and emission spectra, fluorescence quantum yields (Φ F) and lifetimes. Finally, to appraise in depth the deactivation of the excited state of the keto form, femtosecond transient absorption (TA) experiments and Density Functional Theory (DFT) and Time Dependent (TD)-DFT calculations were performed. In an organic aprotic solvent (N,N-dimethylformamide), TA experiments showed a fast deactivation channel (τ 1 = 2.9 ps), which was ascribed to solvent reorganization, and a longer decay component (τ 2 = 86 ps) associated with an internal conversion (IC) process to the ground-state, in opposition to the excited state proton transfer (ESPT) mechanism that takes place in the indigo molecules but in protic solvents. A comparative study was also carried out on the parent molecule, 7,7'-diazaindigo, corroborating the previous conclusions obtained for the alkyl derivative. In agreement with experimental observations, DFT and TD-DFT calculations revealed that the deactivation of the S1 state of the keto form takes place through an internal conversion process.


A Human-Algorithm Integration System for Hip Fracture Detection on Plain Radiography: System Development and Validation Study.
Hip fracture is the most common type of fracture in elderly individuals. Numerous deep learning (DL) algorithms for plain pelvic radiographs (PXRs) have been applied to improve the accuracy of hip fracture diagnosis. However, their efficacy is still undetermined. The objective of this study is to develop and validate a human-algorithm integration (HAI) system to improve the accuracy of hip fracture diagnosis in a real clinical environment. The HAI system with hip fracture detection ability was developed using a deep learning algorithm trained on trauma registry data and 3605 PXRs from August 2008 to December 2016. To compare their diagnostic performance before and after HAI system assistance using an independent testing dataset, 34 physicians were recruited. We analyzed the physicians' accuracy, sensitivity, specificity, and agreement with the algorithm; we also performed subgroup analyses according to physician specialty and experience. Furthermore, we applied the HAI system in the emergency departments of different hospitals to validate its value in the real world. With the support of the algorithm, which achieved 91% accuracy, the diagnostic performance of physicians was significantly improved in the independent testing dataset, as was revealed by the sensitivity (physician alone, median 95%; HAI, median 99%; P<.001), specificity (physician alone, median 90%; HAI, median 95%; P<.001), accuracy (physician alone, median 90%; HAI, median 96%; P<.001), and human-algorithm agreement [physician alone κ, median 0.69 (IQR 0.63-0.74); HAI κ, median 0.80 (IQR 0.76-0.82); P<.001. With the help of the HAI system, the primary physicians showed significant improvement in their diagnostic performance to levels comparable to those of consulting physicians, and both the experienced and less-experienced physicians benefited from the HAI system. After the HAI system had been applied in 3 departments for 5 months, 587 images were examined. The sensitivity, specificity, and accuracy of the HAI system for detecting hip fractures were 97%, 95.7%, and 96.08%, respectively. HAI currently impacts health care, and integrating this technology into emergency departments is feasible. The developed HAI system can enhance physicians' hip fracture diagnostic performance.


Identifying unmet needs in older patients--nurse-GP collaboration in general practice.
Australia's rapidly aging population has a high prevalence of chronic disease and disability, leading to an increased social and economic burden. The Enhanced Primary Care program seeks to reduce this burden by promoting preventive and coordinated care. This study aimed to identify unmet needs in community dwelling general practice patients aged 75 years and over through annual health assessments performed by a general practitioner-nurse team. Community dwelling patients of a large suburban general practice aged 75 years and over were invited to participate. Five hundred and forty-six consecutive, eligible patients were recruited. Data were collected by GP-nurse teams on physical and psychosocial variables using a combination of physical examination, self reporting, and rating scales. Fifty percent of the women and 25% of the men lived alone. Over 90% of participants reported one or more health problems, with musculoskeletal issues being most common. Men rated their health more poorly than women. Incontinence affected one-third of patients, mainly women. Women reported more psychological distress. There were age and gender differences in activities of daily living (ADL). Mobility, ADL, visual impairment, bowel problems, use of sleep medications and psychological wellbeing were strongly associated to self reported health. Health assessments were effective in identifying significant physical and psychosocial problems in older adults. The importance of such assessments is underscored by strong associations between various domains and perceived general health. Collaboration between a GP and a practice based community nurse represents a potential solution to identifying (and responding to) unmet physical and psychosocial needs to improve quality of life in community dwelling older adults.


Leaf energy balance modelling as a tool to infer habitat preference in the early angiosperms.
Despite more than a century of research, some key aspects of habitat preference and ecology of the earliest angiosperms remain poorly constrained. Proposed growth ecology has varied from opportunistic weedy species growing in full sun to slow-growing species limited to the shaded understorey of gymnosperm forests. Evidence suggests that the earliest angiosperms possessed low transpiration rates: gas exchange rates for extant basal angiosperms are low, as are the reconstructed gas exchange rates for the oldest known angiosperm leaf fossils. Leaves with low transpirational capacity are vulnerable to overheating in full sun, favouring the hypothesis that early angiosperms were limited to the shaded understorey. Here, modelled leaf temperatures are used to examine the thermal tolerance of some of the earliest angiosperms. Our results indicate that small leaf size could have mitigated the low transpirational cooling capacity of many early angiosperms, enabling many species to survive in full sun. We propose that during the earliest phases of the angiosperm leaf record, angiosperms may not have been limited to the understorey, and that some species were able to compete with ferns and gymnosperms in both shaded and sunny habitats, especially in the absence of competition from more rapidly growing and transpiring advanced lineages of angiosperms.


"Mixing" as an ethnoetiology of HIV/AIDS in Malaysia's multinational factories.
Minah Karan, the stigmatizing label appended to Malay factory women in the 1980s, signaled a dangerous female sexuality that risked spreading beyond the factory gates and infecting Malaysia's idea(l)s of its traditional kampung culture. This article narrates how Minah Karan, as the former antihero of development, was reconstituted in the 1990s, with the government's labeling of factories as "high-risk settings" for HIV/AIDS. This is an ethnoetiology based not on any evidential epidemiological data but on the racial and gendered "mixing" that transpires behind factory walls: a fear that the "mixing of the sexes" means ipso facto "sexual mixing" among the races. The article demonstrates how importation of the high-risk label articulates at the local level the new and contested linkages, economic, religious, and scientific, constitutive of globalization. The pragmatic nature and imperatives of this high-risk process are discerned in factory women's accounts of how they negotiate the interactional imperatives of factory work, because transnational structures of productivity violate the social boundaries that have long connoted political stability, moral integrity, ethnic community, and individual safety. The article concludes by questioning whether ethnoetiologies, especially when they concern sexual networks, become social etiologies, because this would locate ethnoetiologies as central to conventional public health praxis rather than as ethnographic exotica in the margins.


Vagal control of heart rate is modulated by extracellular potassium.
Heart rate (HR) recovery from heavy exercise is associated with a shift in cardiac sympatho-vagal balance and a transient hypokalaemia. Since changes in extracellular potassium ([K+]0) affect membrane currents in the sino-atrial node, in particular the acetylcholine-activated potassium current (I(K,ACh)), the hyperpolarization-activated current (I(f)) and the L-type calcium current (I(Ca,L)), we investigated whether mimicking [K+]0 concentrations seen during and immediately after exercise could directly modulate the HR response to vagal nerve stimulation (VNS) in the isolated guinea-pig atria preparation pre-stimulated with noradrenaline (NA, 1 microM). Lowering [K+]0 from 4 to 3 mM significantly enhanced the HR response to VNS (5 Hz, 5 V, 30 s, deltaHR 84.5 +/- 14.1 bpm and 119.3 +/- 18.2 bpm, respectively). Increasing [K+]0 to 8 or 10 mM significantly decreased the drop in HR with VNS in comparison to the response to 3 mM K+ Tyrode (deltaHR 56.4 +/- 9.1 bpm and 52.1 +/- 8.7 bpm, respectively). These results could be simulated using the OXSOFT heart sino-atrial node computer model by activating I(K,ACh) during changes in [K+]0. However, changing [K+]0 in the model had no significant effect on the decrease in beating frequency brought about by decreasing I(f) or I(Ca,L). We conclude that the magnitude of the decrease in HR with VNS is enhanced in low [K +]0 and reduced in high [K+]0. The increased efficacy of cardiac vagal activation in low [K+]0 might therefore facilitate the drop in HR after heavy exercise where there is a transient hypokalaemia. Modelling suggests this result may be explained by the effects of changes in [K+]0 on the current-voltage relationship for I(K,ACh).


Overt attentional correlates of memorability of scene images and their relationships to scene semantics.
Computer vision-based research has shown that scene semantics (e.g., presence of meaningful objects in a scene) can predict memorability of scene images. Here, we investigated whether and to what extent overt attentional correlates, such as fixation map consistency (also called inter-observer congruency of fixation maps) and fixation counts, mediate the relationship between scene semantics and scene memorability. First, we confirmed that the higher the fixation map consistency of a scene, the higher its memorability. Moreover, both fixation map consistency and its correlation to scene memorability were the highest in the first 2 seconds of viewing, suggesting that meaningful scene features that contribute to producing more consistent fixation maps early in viewing, such as faces and humans, may also be important for scene encoding. Second, we found that the relationship between scene semantics and scene memorability was partially (but not fully) mediated by fixation map consistency and fixation counts, separately as well as together. Third, we found that fixation map consistency, fixation counts, and scene semantics significantly and additively contributed to scene memorability. Together, these results suggest that eye-tracking measurements can complement computer vision-based algorithms and improve overall scene memorability prediction.


Preoperative Predictors of Achieving Clinically Significant Athletic Functional Status After Hip Arthroscopy for Femoroacetabular Impingement at Minimum 2-Year Follow-Up.
To identify predictors of achieving clinically significant sport function in athletic patients undergoing hip arthroscopy for femoroacetabular impingement syndrome (FAIS). Data were analyzed for all patients who treated for FAIS between 2012 to 2016 and reported being athletes, including recreational and competitive athletes. All patients had a minimum of 2-year follow-up with patient-reported athletic function in the form of the Hip Outcome Score-Sport Specific (HOS-SS), visual analog score-pain, and patient satisfaction. Achieving clinically significant sports function was defined as either reaching the minimally clinical important difference (MCID) or the patient acceptable symptomatic state (PASS) for HOS-SS at 2-year follow-up. An exploratory factor analysis was used to determine specific domains for the predictor variables and to reduce the redundancy in these variables. A logistic regression analysis was used to identify significant predictors of achieving clinically significant sports function [corrected]. Of 780 qualifying patients, 626 completed the 2-year minimum follow-up (80%), with a mean age and body mass index of 31.6 ± 11.9 years and 24.6 ± 8.6, respectively. A total of 500 patients (86.5%) achieved high functional status, with 77.9% achieving MCID HOS-SS and 68.7% achieving PASS HOS-SS. Logistic regression analysis identified increased the α angle (odds ratio [OR] 0.976; P = .027), preoperative pain duration (OR 0.729; P = .011), and body mass index (BMI) (OR 0.919; P = .018), as well as the presence of femoral chondral defects (OR 0.769; P = .013), as negative predictors for achieving MCID. Negative predictors for achieving PASS HOS-SS included the presence of a preoperative limp (OR 0.384; P = .013), anxiety or depression (OR 0.561; P = .041), and increased BMI (OR 0.945; P = .018) and preoperative pain duration (OR 0.987; P < .001). Several predictors of achieving clinically significant sport function performance exist, including a history of anxiety or depression, BMI, preoperative α angle, limp, femoral chondral damage, *and preoperative symptom duration. Our results suggest there are both modifiable and nonmodifiable preoperative factors that have the potential to predict achieving high athletic function after hip arthroscopy for FAIS. IV, Case Series.


HIV testing and receipt of test results among homeless persons with serious mental illness.
The purpose of this study was to determine the rates and predictors of HIV testing and receipt of results among homeless adults with serious mental illness in the initial 3-month period after contact with a community-based case management program. Baseline and follow-up interview data came from clients (N=5,890) in the Access to Community Care and Effective Services and Supports program, an 18-site, 5-year federally sponsored demonstration designed to evaluate the effect of service system integration on outcomes for homeless persons with serious mental illness. Overall, 38.0% of clients were tested for HIV in the 3 months after program entry; of these, 88.8% returned to receive their test results. Likelihood of being tested was independently associated with having been tested before, more severe psychiatric symptoms and drug problems, level of worry about getting AIDS, younger age, less education, minority status, longer-term homelessness, being sexually assaulted, being arrested, and health services utilization. Among those tested, likelihood of receiving the test results was higher among those with a history of prior testing and return for results, a higher frequency of testing, and more years of education and lower among those with drug abuse problems, outpatient medical service utilization, disability, and sexually transmitted disease. Interaction analyses showed that, for men, greater social support increased the likelihood of both HIV testing and receipt of results, while sexual victimization during follow-up decreased the likelihood that men would return for their HIV results. The majority of homeless clients enrolled in an intensive case management program were not tested for HIV during the 3-month period after program entry. Among those tested, however, nearly 90% reported receiving their results. The findings may enhance the development and targeting of strategies to increase testing and awareness of HIV serostatus among high-risk mentally ill homeless persons.


Study of diverse mechanisms of cell-mediated cytotoxicity in gene-targeted mice using flow cytometric cytotoxicity assay.
Cytotoxic lymphocytes kill tumor or virus-infected target cells utilizing two mechanisms-(1) release of lytic granules (containing perforin and granzymes), and (2) Fas ligand (FasL)/Fas or TNF-initiated apoptosis. We have examined mechanisms of target cell lysis by effector T cells from gene-targeted and mutant mice using a new Flow Cytometric Cytotoxicity Assay (FC Assay). Target cells were labeled with PKH67 dye. Cell death was estimated by 7-AAD inclusion and annexin V-PE binding. A direct correlation has been found between the percentage of dead target cells in FC Assay and the results of 111In release cytotoxicity assay when effector T cells from either Pfp -/- (perforin knockout) or gld (non-functional Fas Ligand) mice were used. As shown by the 4 h FC assay, the granule-mediated mechanism was utilized by T cells from gld mice. In contrast, T cells from Pfp -/- mice used death receptor-mediated lysis. Therefore, cytotoxic cells from gene-targeted and mutant mice can serve as valuable tools for studying different mechanisms of cell-mediated cytotoxicity, and the FC assay could be applied irrespective of which cytotoxic effector pathway is involved.


Chronotropic and pressor effects of water ingestion at rest and in response to incremental dynamic exercise.
Ingestion of water attenuates the chronotropic response to submaximal exercise. However, it is not known whether this effect is equally manifested during dynamic exercise below and above the ventilatory threshold (VT). We explored the effects of water ingestion on the heart rate response to an incremental cycle-ergometer protocol. In a randomized fashion, 19 healthy adults (10 men and nine women, age 20.9 ± 1.8 years) ingested 50 and 500 ml of water before completing a cycle-ergometer protocol on two separate days. The heart rate and oxygen uptake ( ) responses to water ingestion were analysed both at rest and during exercise performed below and above the VT. The effects of water intake on brachial blood pressure were measured only at rest. Resting mean arterial pressure increased and resting heart rate decreased, but only after 500 ml of water (P < 0.05). Compared with that seen after 50 ml of water, the 500 ml volume elicited an overall decrease in submaximal heart rate (P < 0.05). In contrast, drinking 500 ml of water did not affect submaximal . The participants' maximal heart rate, maximal and VT were similar between conditions. Our results therefore indicate that, owing to its effects on submaximal heart rate over a broad spectrum of intensities, the drinking of water should be recognized as a potential confounder in cardiovascular exercise studies. However, by showing no differences between conditions for submaximal , they also suggest that the magnitude of heart rate reduction after drinking 500 ml of water may be of minimal physiological significance for exercise cardiorespiratory capacity.


A single-cell map of intratumoral changes during anti-PD1 treatment of patients with breast cancer.
Immune-checkpoint blockade (ICB) combined with neoadjuvant chemotherapy improves pathological complete response in breast cancer. To understand why only a subset of tumors respond to ICB, patients with hormone receptor-positive or triple-negative breast cancer were treated with anti-PD1 before surgery. Paired pre- versus on-treatment biopsies from treatment-naive patients receiving anti-PD1 (n = 29) or patients receiving neoadjuvant chemotherapy before anti-PD1 (n = 11) were subjected to single-cell transcriptome, T cell receptor and proteome profiling. One-third of tumors contained PD1-expressing T cells, which clonally expanded upon anti-PD1 treatment, irrespective of tumor subtype. Expansion mainly involved CD8+ T cells with pronounced expression of cytotoxic-activity (PRF1, GZMB), immune-cell homing (CXCL13) and exhaustion markers (HAVCR2, LAG3), and CD4+ T cells characterized by expression of T-helper-1 (IFNG) and follicular-helper (BCL6, CXCR5) markers. In pre-treatment biopsies, the relative frequency of immunoregulatory dendritic cells (PD-L1+), specific macrophage phenotypes (CCR2+ or MMP9+) and cancer cells exhibiting major histocompatibility complex class I/II expression correlated positively with T cell expansion. Conversely, undifferentiated pre-effector/memory T cells (TCF7+, GZMK+) or inhibitory macrophages (CX3CR1+, C3+) were inversely correlated with T cell expansion. Collectively, our data identify various immunophenotypes and associated gene sets that are positively or negatively correlated with T cell expansion following anti-PD1 treatment. We shed light on the heterogeneity in treatment response to anti-PD1 in breast cancer.


Interaction of water with oligo(ethylene glycol) terminated monolayers: wetting versus hydration.
Biorepulsivity of oligo(ethylene glycol) (OEG) substituted self-assembled monolayers (SAMs), serving as model systems for analogous polymeric surfaces, is generally ascribed to the hydration effect. In this context, we applied temperature-programmed desorption to study interaction of water (D2O) with a series of OH-terminated, OEG-substituted alkanethiolate SAMs with variable length of the OEG strand, defining their biorepulsion behavior. Along with the ice overlayer (wetting phase), growing also on the surface of the analogous non-substituted films, a hydration phase, corresponding to the adsorption of D2O into the OEG matrix, was observed, with a higher desorption energy (12.4 kcal mol-1vs. 10.4 kcal mol-1) and a weight correlating with the length of the OEG strand and, consequently, with biorepulsivity. The formation of hydration phase was found to occur over an activation barrier, presumably by temperature-promoted diffusion from the wetting phase, with this process being additionally enforced by a pre-desorption annealing.


Establishing benchmark rates of microbial and bacterial endotoxin contamination for radiopharmaceuticals compounded in commercial nuclear pharmacy settings.
The objective of this study was to establish benchmark rates for microbial and bacterial endotoxin contamination rates for radiopharmaceutical preparations compounded in commercial nuclear pharmacies. Radiopharmaceutical samples were obtained between November 2006 and June 2010 from seven commercial nuclear pharmacies. Preparations were compounded per the compounding protocols of each radiopharmacy, and each kit was used for unit-dose dispensing of patient-specific doses. Samples for testing were withdrawn after unit doses were dispensed. Sterility testing was performed on each radiopharmaceutical sample and incubated at appropriate temperatures for 14 days. A sample of the radiopharmaceutical was also used to complete limulus amebocyte lysate-based bacterial endotoxin testing. Over the course of the study, 1516 radiopharmaceutical samples from 16 different radiopharmaceutical preparations, including eluates from radionuclide generators, were tested for sterility and bacterial endotoxicity. For sterility testing, 13 of the 1516 samples (0.86%) showed evidence of growth in the testing media, indicating the presence of microbes in the tested sample. For bacterial endotoxin testing, 4 or 1492 samples (0.27% showed formation of gel clots, indicating the presence of bacterial endotoxins in the sample. The microbial and bacterial endotoxin contamination rates of aseptically compounded radiopharmaceuticals compounded in a commercial nuclear pharmacy environment are extremely low. The results of this study show the high level of safety and quality that is provided when obtaining radiopharmaceutical doses that are compounded and dispensed from a commercial nuclear pharmacy.


Does morphological structure modulate access to embedded word meaning in child readers?
Beginning readers have been shown to be sensitive to the meaning of embedded neighbors (e.g., CROW in CROWN). Moreover, developing readers are sensitive to the morphological structure of words (TEACH-ER). However, the interaction between orthographic and morphological processes in meaning activation during reading is not well established. What determines semantic access to orthographically embedded words? What is the role of suffixes in this process? And how does this change throughout development? To address these questions, we asked 80 Italian elementary school children (third, fourth, and fifth grade) to make category decisions on words (e.g., is CARROT a type of food?). Critically, some target words for no-answers (e.g., is CORNER a type of food?) contained category-congruent embedded stems (i.e., CORN). To gauge the role of morphology in this process, half of the embedded stems were accompanied by a pseudosuffix (CORN-ER) and half by a non-morphological ending (PEA-CE). Results revealed that words were harder to reject as members of a category when the embedded stem was category-congruent. This effect held both with and without a pseudosuffix, but was larger for pseudosuffixed words in the error rates. These results suggest that orthographic stems are activated and activation is fed forward to the semantic level regardless of morphological structure, followed by a decision-making process that might strategically use suffix-like endings.


Otologic manifestations of malignant osteopetrosis.
To determine the incidence of hearing loss and describe the neurotologic manifestations over time in a large series of patients with malignant osteopetrosis. Longitudinal study. Tertiary care referral center. Thirty-two patients, including 19 infants (< 1 yr of age at initial visit) and 13 children (aged 1-7.6 yr at initial visit), with autosomal recessive osteopetrosis were followed-up during a 10-year period from 1991 to 2001. The average length of follow-up was 2.5 years (range, 0-9.1 yr). Patients underwent annual otologic evaluations including clinical examination, audiologic evaluation (auditory brainstem response, pure-tone thresholds, speech discrimination scores, and tympanograms), and high-resolution computed tomography of the temporal bone. Incidence of hearing loss, otitis media, and facial paralysis. Serial changes in temporal bone anatomy by computed tomographic scan. Twenty-six percent of infants' ears demonstrated hearing loss during the first year of life, and 78% of children's ears demonstrated hearing loss during the study period. Of the children's ears with hearing loss, 100% had a conductive component and 26% had an additional sensorineural component (mixed hearing loss); VIIIth nerve conduction was normal in 100% of infants and 78% of children. Five patients (16%) had unilateral facial nerve palsy. Common temporal bone findings included thickening and sclerosis of the calvarium; poor pneumatization of the mastoid bone; and narrowing of the external auditory canal, eustachian tube, and internal auditory canal. Otologic manifestations are common in malignant osteopetrosis secondary to the formation of dense, brittle bone. Frequent findings include external auditory canal stenosis, otitis media, conductive and sensorineural hearing loss, and facial nerve paralysis.


npInv: accurate detection and genotyping of inversions using long read sub-alignment.
Detection of genomic inversions remains challenging. Many existing methods primarily target inzversions with a non repetitive breakpoint, leaving inverted repeat (IR) mediated non-allelic homologous recombination (NAHR) inversions largely unexplored. We present npInv, a novel tool specifically for detecting and genotyping NAHR inversion using long read sub-alignment of long read sequencing data. We benchmark npInv with other tools in both simulation and real data. We use npInv to generate a whole-genome inversion map for NA12878 consisting of 30 NAHR inversions (of which 15 are novel), including all previously known NAHR mediated inversions in NA12878 with flanking IR less than 7kb. Our genotyping accuracy on this dataset was 94%. We used PCR to confirm the presence of two of these novel inversions. We show that there is a near linear relationship between the length of flanking IR and the minimum inversion size, without inverted repeats. The application of npInv shows high accuracy in both simulation and real data. The results give deeper insight into understanding inversion.


Itch and skin rash from chocolate during fluoxetine and sertraline treatment: case report.
The skin contains a system for producing serotonin as well as serotonin receptors. Serotonin can also cause pruritus when injected into the skin. SSRI-drugs increase serotonin concentrations and are known to have pruritus and other dermal side effects. A 46-year-old man consulted his doctor due to symptoms of depression. He did not suffer from any allergy but drinking red wine caused vasomotor rhinitis. Antidepressive treatment with fluoxetine 20 mg daily was initiated which was successful. After three weeks of treatment an itching rash appeared. An adverse drug reaction (ADR) induced by fluoxetine was suspected and fluoxetine treatment was discontinued. The symptoms disappeared with clemastine and betametasone treatment. Since the depressive symptoms returned sertraline medication was initiated. After approximately two weeks of sertraline treatment he noted an intense itching sensation in his scalp after eating a piece of chocolate cake. The itch spread to the arms, abdomen and legs and the patient treated himself with clemastine and the itch disappeared. He now realised that he had eaten a chocolate cake before this episode and remembered that before the first episode he had had a chocolate mousse dessert. He had never had any reaction from eating chocolate before and therefore reported this observation to his doctor. This case report suggests that there may be individuals that are very sensitive to increases in serotonin concentrations. Dermal side reactions to SSRI-drugs in these patients may be due to high activity in the serotonergic system at the dermal and epidermo-dermal junctional area rather than a hypersensitivity to the drug molecule itself.


Activation of Pyk2 by stress signals and coupling with JNK signaling pathway.
The c-Jun amino-terminal kinase (JNK) is activated by various heterotrimeric guanine nucleotide-binding protein (G protein)-coupled receptors, inflammatory cytokines, and stress signals. Yet, upstream mediators that link extracellular signals with the JNK signaling pathway are currently unknown. The tyrosine kinase Pyk2 was activated by tumor necrosis factor alpha, by ultraviolet irradiation, and by changes in osmolarity. Overexpression of Pyk2 led to activation of JNK, and a dominant-negative mutant of Pyk2 interfered with ultraviolet light- or osmotic shock-induced activation of JNK. Pyk2 represents a cell type-specific, stress-sensitive mediator of the JNK signaling pathway.


The relationship between bottle usage/content, age, and number of teeth with mutans streptococci colonization in 6-24-month-old children.
Mutans streptococci (MS) are the primary pathogens involved in the development of early childhood caries. However, factors that may affect their acquisition in the mouths of young children are not well understood, and the period of initial colonization remains controversial. This study investigated the relationship of age, number of teeth, and bottle usage/content with regard to the isolation of MS in 6-24-month-old children. A total of 122 children from low-income families attending a nutritional supplement program, and their mothers, participated in this study. Children were examined for dental caries and number of erupted teeth and were sampled for MS. Mothers were administered a questionnaire to obtain details of baby bottle use, including what food items were put in the bottle during the last week. MS was detected in more than one-third of the 6-24-month-olds. Unlike some studies that suggest a later period of infectivity, approximately 20% of children under 14 months of age, including 4 of 22 infants aged 6-9 months, were colonized with MS. When examined separately, age, number of teeth, and bottle usage/content were each found to be related to the presence of MS. Mutans streptococci colonization was more likely with increasing age and number of teeth, and children whose bottles contained sweetened beverages were more likely to be colonized than children whose bottles contained milk. Logistic regression models that controlled for both age and number of teeth indicated that children who consumed sweetened beverages in their baby bottle had a statistically significant, four-fold increase in the odds of colonization by MS relative to children who consumed milk. The finding that approximately 20% of the children under 14 months of age were infected with MS indicates that colonization in this sample of low-income preschool children may begin earlier than suggested by some investigations. Additionally, the risk of MS colonization appears lower among infants who consume milk rather than sweetened beverages in the bottle.


Improved metabolic regulation is associated with retinoblastoma protein gene haploinsufficiency in mice.
Retinoblastoma protein (pRb) is involved in the control of energy metabolism, and its inactivation protects mice against high-fat diet-induced diabesity. Here, we tested the hypothesis that partial deficiency in the Rb gene could confer metabolic advantages in front of acute challenges to metabolism and as mice age on a regular diet. Rb haploinsufficient (Rb(+/-)) mice and wild-type (WT) littermates were studied from weaning and characterized at 1.5-2.5 mo of age (young adults) and 6-7.5 mo of age (mature adults). Whereas no differences in body weight or composition were observed at young age, mature adult Rb(+/-) mice were leaner than WT littermates, displaying 36% reduced body fat content. At both ages studied, Rb(+/-) mice displayed improved blood lipids, enhanced sensitivity to the blood glucose-lowering effect of insulin and to the anorectic effect of leptin, and a reduced respiratory exchange ratio, indicative of an increased use of fatty acids as a fuel. Insulin sensitivity and oral fat tolerance were better maintained with age in the Rb(+/-) than the WT mice. Mature adult Rb(+/-) mice displayed gene expression changes consistent with increased fatty acid oxidation in white adipose tissue and skeletal muscle and paramount signs of browning in the inguinal white adipose tissue. In conclusion, Rb haploinsufficiency provides metabolic advantages in front of acute metabolic stressors and ameliorates body fat gain and metabolic impairments that normally accompany transition from young to mature adult age.


A fibrous membrane suspends the multifocal lens in the eyes of lampreys and African lungfishes.
The sharpness and thus information content of the retinal image in the eye depends on the optical quality of the lens and its accurate positioning in the eye. Multifocal lenses create well-focused color images and are present in the eyes of all vertebrate groups studied to date (mammals, reptiles including birds, amphibians, and ray-finned fishes) and occur even in lampreys, i.e., the most basal vertebrates with well-developed eyes. Results from photoretinoscopy obtained in this study indicate that the Dipnoi (lungfishes), i.e., the closest piscine relatives to tetrapods, also possess multifocal lenses. Suspension of the lens is complex and sophisticated in teleosts (bony fishes) and tetrapods. We studied lens suspension using light and electron microscopy in one species of lamprey (Lampetra fluviatilis) and two species of African lungfish (Protopterus aethiopicus aethiopicus and Protopterus annectens annectens). A fibrous and highly transparent membrane suspends the lens in both of these phylogenetically widely separated vertebrate groups. The membrane attaches to the lens approximately along the lens equator, from where it extends to the ora retinalis. The material forming the membrane is similar in ultrastructure to microfibrils in the zonule fibers of tetrapods. The membrane, possibly in conjunction with the cornea, iris, and vitreous body, seems suitable for keeping the lens in the correct position for well-focused imaging. Suspension of the lens by a multitude of zonule fibers in tetrapods may have evolved from a suspensory membrane similar to that in extant African lungfishes, a structure that seems to have appeared first in the lamprey-like ancestors of allextant vertebrates.


Improvement in mycobacterial yield and reduced time to detection in pediatric samples by use of a nutrient broth growth supplement.
There is an urgent need to improve the methods used for the bacteriological diagnosis of childhood mycobacterial disease. This study compared the mycobacterial yields and the times to detection (in days) of mycobacteria in pediatric clinical specimens by using Mycobacterial Growth Indicator Tubes (MGITs) and solid Löwenstein-Jensen (LJ) slants with and without a nutrient broth supplement. A total of 801 specimens from 493 patients were processed: 82.8% were gastric aspirate specimens, 15.6% were sputum specimens, and 1.6% were fine-needle-aspiration biopsy specimens. The mycobacterial yield obtained with MGITs (with and without nutrient broth) was 11.0%, and that obtained with LJ slants was 1.6% (P < 0.001). Of the 88 positive cultures, 62 were detected in MGITs and 73 were detected in MGITs supplemented with nutrient broth (P = 0.11). The mean time to detection in MGITs (without nutrient broth) was 18.5 days, whereas it was 12.4 days in MGITs with nutrient broth (P < 0.001). Supplementation of standard MGITs improved the mycobacterial yield and significantly reduced the time to detection of mycobacteria in pediatric samples.


Pancreatic β-cell identity, glucose sensing and the control of insulin secretion.
Insulin release from pancreatic β-cells is required to maintain normal glucose homoeostasis in man and many other animals. Defective insulin secretion underlies all forms of diabetes mellitus, a disease currently reaching epidemic proportions worldwide. Although the destruction of β-cells is responsible for Type 1 diabetes (T1D), both lowered β-cell mass and loss of secretory function are implicated in Type 2 diabetes (T2D). Emerging results suggest that a functional deficiency, involving de-differentiation of the mature β-cell towards a more progenitor-like state, may be an important driver for impaired secretion in T2D. Conversely, at least in rodents, reprogramming of islet non-β to β-cells appears to occur spontaneously in models of T1D, and may occur in man. In the present paper, we summarize the biochemical properties which define the 'identity' of the mature β-cell as a glucose sensor par excellence. In particular, we discuss the importance of suppressing a group of 11 'disallowed' housekeeping genes, including Ldha and the monocarboxylate transporter Mct1 (Slc16a1), for normal nutrient sensing. We then survey the changes in the expression and/or activity of β-cell-enriched transcription factors, including FOXO1, PDX1, NKX6.1, MAFA and RFX6, as well as non-coding RNAs, which may contribute to β-cell de-differentiation and functional impairment in T2D. The relevance of these observations for the development of new approaches to treat T1D and T2D is considered.


Who benefits from psychotherapies for adult depression? A meta-analytic update of the evidence.
It is not clear whether specific target groups for psychotherapies in adult depression benefit as much from these treatments as other patients. We examined target groups that have been examined in randomized trials, including women, older adults, students, minorities, patients with general medical disorders, and specific types of depression, and we examined where patients were recruited. We conducted subgroup and multivariate metaregression analyses in a sample of 256 trials (with 332 comparisons) comparing psychotherapy with an inactive control condition. Only 22% of the studies had low risk of bias (RoB), heterogeneity was high and there was a considerable risk of publication bias. A meta-regression analysis among low RoB studies showed that effect sizes found for studies among women, older adults, patients with general medical disorders, patients recruited from primary care, and patients scoring above a cut-off on a self-rating depression scale, did not differ significantly from effect sizes from other studies. For other target groups, the number of low RoB studies was too small to draw any conclusion. We found few indications that psychotherapies for adult depression are more or less effective in women, older adults, patients with comorbid general medical disorders, and primary care patients.


Importance of Decision Support Implementation in Emergency Department Vancomycin Dosing.
The emergency department (ED) plays a critical role in the management of life-threatening infection. Prior data suggest that ED vancomycin dosing is frequently inappropriate. The objective is to assess the impact of an electronic medical record (EMR) intervention designed to improve vancomycin dosing accuracy, on vancomycin dosing and clinical outcomes in critically ill ED patients. Retrospective before-after cohort study of all patients (n=278) treated with vancomycin in a 60,000-visit Midwestern academic ED (March 2008 and April 2011) and admitted to an intensive care unit. The primary outcome was the proportion of vancomycin doses defined as "appropriate" based on recorded actual body weight. We also evaluated secondary outcomes of mortality and length of stay. The EMR dose calculation tool was associated with an increase in mean vancomycin dose ([14.1±5.0] vs. [16.5±5.7] mg/kg, p<0.001) and a 10.3% absolute improvement in first-dose appropriateness (34.3% vs. 24.0%, p=0.07). After controlling for age, gender, methicillin-resistant staphylococcus aureus infection, and Acute Physiology and Chronic Health Evaluation II score, 28-day in-hospital mortality (odds ratio OR 1.72; 95% CI [0.76-3.88], p=0.12) was not affected. A computerized decision-support tool is associated with an increase in mean vancomycin dose in critically ill ED patients, but not with a statistically significant increase in therapeutic vancomycin doses. The impact of decision-support tools should be further explored to optimize compliance with accepted antibiotic guidelines and to potentially affect clinical outcome.


Diversity of bacteria associated with grassland soil nematodes of different feeding groups.
The bacterial diversity associated with soil nematodes and its relationship with their feeding habits are as yet poorly understood. In the present study the diversity and abundance of bacteria from nematodes and their surrounding soil were analysed and compared. The nematodes were collected from a grassland soil and sorted into bacterial, fungal, plant, predatory and omnivore feeding groups and assigned to taxonomic groups. Total DNA was extracted from the nematodes and partial bacterial 16S rRNA genes were PCR amplified, cloned and sequenced. The abundance and composition of bacterial taxa differed between and within feeding groups. The lowest bacterial diversity was found in the predatory nematodes Prionchulus sp., whereas the highest bacterial diversity was associated with the bacterial-feeding nematode Acrobeles sp. The soil had a more diverse bacterial community than the communities found in the nematode groups. The 16S rRNA gene sequences of bacteria associated with nematodes did not overlap with those detected in soil as determined using the cloning screening approach. However, bacterial sequences identified from nematodes could be detected in the soil with targeted PCR. Our data suggest that the nematodes do not feed on the most abundant bacteria present in soil. Furthermore, several nematodes contained suspected bacterial symbionts and parasites.


Living and dying in East Africa: implementing the End-of-Life Nursing Education Consortium curriculum in Tanzania.
Education about palliative care is crucial for oncology nurses, particularly in the developing world, where most patients present with advanced disease and heavy symptom burden. The End-of-Life Nursing Education Consortium-International training program was implemented in Tanzania to provide nurses with the knowledge, expertise, and tools to better care for the dying and to educate others. The curriculum was presented to 39 participants over three days, including didactic presentations, small group discussions, and role play. None of the participants had received previous formal palliative care training. The participants rated their impression of the course as excellent. Follow-up at eight months yielded similar findings regarding the effectiveness of the curriculum. International education regarding palliative care for those with cancer requires an understanding of different disease patterns and clinical practices, along with cultural humility and empathy. These experiences are extraordinarily enriching, giving nurses a unique perspective on palliative care that ultimately informs their own practice.


Dexmedetomidine Sedation for Paroxysmal Supraventricular Tachycardia Ablation Is Not Associated With Alteration of Arrhythmia Inducibility.
Dexmedetomidine (Dex) is an attractive agent for procedural sedation due to its unique pharmacodynamic profile, specifically affording predictable sedation without concurrent respiratory depression. However, Dex has previously been reported to prevent or terminate arrhythmias. The purpose of this study was to investigate paroxysmal supraventricular tachycardia (PSVT) inducibility and homeostatic stability during electrophysiology studies (EPSs) and ablation when a standardized Dex protocol was used as the primary sedation agent. We performed a retrospective review of 163 consecutive procedures for PSVT ablation that received Dex as the primary sedative with adjunct fentanyl and midazolam boluses (DEX-FENT-MIDAZ). This cohort was compared to 163 consecutive control procedures wherein strictly fentanyl and midazolam were used for sedation. The primary outcome reviewed was PSVT inducibility assessed before ablation. Reviewed secondary outcomes included level of sedation and intraprocedure hemodynamics and oxygenation. The arrhythmia profiles of the DEX-FENT-MIDAZ and control cohorts were very similar. The overall incidence of a "negative" EPSs in which arrhythmia was not induced was 24% in the DEX-FENT-MIDAZ group and 26% in the control group (P = .7). Unintended deep sedation was significantly less with DEX-FENT-MIDAZ (4.3% vs 27%; P ≤ .0001). However, DEX-FENT-MIDAZ use was associated with a higher incidence of intraprocedure hypotension. Dex sedation during EPSs is not associated with a reduction in PSVT inducibility. The therapeutic utility of Dex during EPS arises from the predictable sedation Dex affords but is associated with an increased incidence of intraprocedure hypotension.


The effect of dopamine agonists on metabolic variables in adults with type 2 diabetes: A systematic review with meta analysis and trial sequential analysis of randomized clinical trials.
To assess the metabolic effects of dopamine agonists compared with placebo in randomized controlled trials (RCTs) including adults with type 2 diabetes. Eligible trials were identified by searching PubMed, Embase and CENTRAL. The primary outcomes were HbA1c and serious adverse events (SAEs) assessed at longest available follow-up. Secondary outcomes were fasting plasma glucose, adverse events, body weight, hypoglycaemia and triglycerides. We assessed risk of bias and evaluated the certainty of the evidence with the Grading of Recommendations Assessment, Development and Evaluation (GRADE). Nine RCTs enrolling 3456 participants were included, six of which assessed the effect of bromocriptine, and the other three the effect of cabergoline. Dopamine agonists reduced HbA1c with 0.69 standardized mean difference (95% CI = 0.28 to 1.09; P = .0008; I2 = 80%; GRADE: low) compared with placebo. There was no difference in the effect between bromocriptine and cabergoline. Heterogeneity was partly explained by dosage and study duration, both of which were inversely associated with effect size. Only one large trial reported SAEs and no difference was reported for the risk of an SAE (RR = 0.89; 95% CI = 0.70 to 1.12; P = .32) between active intervention and placebo. Secondary outcomes suggested a decrease in fasting plasma glucose and triglycerides and no effect on the remaining outcomes. Dopamine agonists reduce HbA1c as well as fasting plasma glucose and triglycerides in patients with type 2 diabetes without causing SAEs. These data are based on moderate to low quality evidence thus our confidence in the effect estimates is limited.


New concepts of obstetric and gynecologic infection.
Multiple organisms are usually recovered from patients with acute pelvic inflammatory disease (PID). The sexually transmitted organisms Neisseria gonorrhoeae, Chlamydia trachomatis, and genital mycoplasmas frequently initiate infection. Although aerobic and anaerobic bacteria that constitute the normal vaginal flora are frequent secondary invaders following an initial sexually transmitted infection, these bacteria can also primarily cause PID. Early postpartum endometritis, the most common postpartum infection, amniotic fluid infection, and infection following hysterectomy are usually caused by the aerobic and anaerobic bacteria that constitute the normal vaginal flora. Because the organisms that cause the various obstetric-gynecologic infections are often found in combination, it is necessary to begin therapy with broad-spectrum antibiotics. Many of these infections are caused by anaerobic bacteria and it is often necessary to select antibiotics that inhibit anaerobes.


Improvement of NADPH bioavailability in Escherichia coli through the use of phosphofructokinase deficient strains.
NADPH-dependent reactions play important roles in production of industrially valuable compounds. In this study, we used phosphofructokinase (PFK)-deficient strains to direct fructose-6-phosphate to be oxidized through the pentose phosphate pathway (PPP) to increase NADPH generation. pfkA or pfkB single deletion and double-deletion strains were tested for their ability to produce lycopene. Since lycopene biosynthesis requires many NADPH, levels of lycopene were compared in a set of isogenic strains, with the pfkA single deletion strain showing the highest lycopene yield. Using another NADPH-requiring process, a one-step reduction reaction of 2-chloroacrylate to 2-chloropropionic acid by 2-haloacrylate reductase, the pfkA pfkB double-deletion strain showed the highest yield of 2-chloropropionic acid product. The combined effect of glucose-6-phosphate dehydrogenase overexpression or lactate dehydrogenase deletion with PFK deficiency on NADPH bioavailability was also studied. The results indicated that the flux distribution of fructose-6-phosphate between glycolysis and the pentose phosphate pathway determines the amount of NAPDH available for reductive biosynthesis.


Q-switched Erbium-doped fiber laser at 1600 nm for photoacoustic imaging application.
We present a nanosecond Q-switched Erbium-doped fiber (EDF) laser system operating at 1600 nm with a tunable repetition rate from 100 kHz to 1 MHz. A compact fiber coupled, acousto-optic modulator-based EDF ring cavity was used to generate a nanosecond seed laser at 1600 nm, and a double-cladding EDF based power amplifier was applied to achieve the maximum average power of 250 mW. In addition, 12 ns laser pulses with the maximum pulse energy of 2.4 μJ were obtained at 100 kHz. Furthermore, the Stokes shift by Raman scattering over a 25 km long fiber was measured, indicating that the laser can be potentially used to generate the high repetition rate pulses at the 1.7 μm region. Finally, we detected the photoacoustic signal from a human hair at 200 kHz repetition rate with a pulse energy of 1.2 μJ, which demonstrates that a Q-switched Er-doped fiber laser can be a promising light source for the high speed functional photoacoustic imaging.


Mannose receptor expression and function define a new population of murine dendritic cells.
In vitro the mannose receptor (MR) mediates Ag internalization by dendritic cells (DC) and favors the presentation of mannosylated ligands to T cells. However, in vivo MR seems to play a role not in Ag presentation but in the homeostatic clearance of endogenous ligands, which could have the secondary benefit of reducing the levels of endogenous Ag available for presentation to the adaptive immune system. We have now observed that while MR(+) cells are consistently absent from T cell areas of spleen and mesenteric lymph nodes (LN), peripheral LN of untreated adult mice contain a minor population of MR(+)MHCII(+) in the paracortex. This novel MR(+) cell population can be readily identified by flow cytometry and express markers characteristic of DC. Furthermore, these MR(+) DC-like cells located in T cell areas can be targeted with MR ligands (anti-MR mAb). Numbers of MR(+)MHCII(+) cells in the paracortex are increased upon stimulation of the innate immune system and, accordingly, the amount of anti-MR mAb reaching MR(+)MHCII(+) cells in T cell areas is dramatically enhanced under these conditions. Our results indicate that the MR can act as an Ag-acquisition system in a DC subpopulation restricted to lymphoid organs draining the periphery. Moreover, the effect of TLR agonists on the numbers of these MR(+) DC suggests that the immunogenicity of MR ligands could be under the control of innate stimulation. In accordance with these observations, ligands highly specific for the MR elicit enhanced humoral responses in vivo only when administered in combination with endotoxin.


Divalent cation ionophores stimulate resorption and inhibit DNA synthesis in cultured fetal rat bone.
Two divalent cation ionophores, A23187 and Ionomycin, which are selective for calcium, stimulated the resorption of fetal rat long bones in organ culture at 0.1 to 1 micromolar but not at higher concentrations. Both agents inhibited DNA synthesis at concentrations that stimulated resorption. These results might explain the differences in ionophore effects on bone previously reported, and they imply that cell replication is not required for osteoclast formation in fetal rat long bone cultures.


Combined effects of noise and gentamicin on hearing in the guinea pig.
The last ten years, the use of gentamicin has increased due to antibiotic resistance among bacterial pathogens. One of the side effects of gentamicin is its toxicity on hearing. Several authors had even pointed out synergistic effects of gentamicin and noise on hearing. It was therefore reasonable to think that the damaging effects of noise could be emphasized by a gentamicin treatment of the subjects. In order to test the applicability of the Leq8h for estimating the hazard of noise on animals treated with a non-ototoxic dose of gentamicin (40 mg/kg for 8 days), two experiments were carried out with guinea pigs. The animals were exposed to octave band noises centred at 8 kHz and treated with gentamicin either simultaneously or sequentially with regard to the noise exposure. Two noise exposures having different acoustic energy, respectively Leq8h = 85 dB and 98.8 dB SPL, were tested. The auditory function of the guinea pigs was tested by recording auditory-evoked potentials. The electrophysiological findings were completed by histological data. The gentamicin treatment tested in the current studies did not cause any auditory permanent threshold shift neither cochlear disruptions, although the treatment could be considered as approximately ten times the therapeutic dose used in human. The auditory deficit induced by the mixed exposures to noise and gentamicin did not worsen the noise effect alone in our experimental conditions. As a result, the European value recommended for noise exposure (Leq8h=85 dB) seems to be robust enough to protect gentamicin-treated workers.


Device and circuit-level performance of carbon nanotube field-effect transistor with benchmarking against a nano-MOSFET.
The performance of a semiconducting carbon nanotube (CNT) is assessed and tabulated for parameters against those of a metal-oxide-semiconductor field-effect transistor (MOSFET). Both CNT and MOSFET models considered agree well with the trends in the available experimental data. The results obtained show that nanotubes can significantly reduce the drain-induced barrier lowering effect and subthreshold swing in silicon channel replacement while sustaining smaller channel area at higher current density. Performance metrics of both devices such as current drive strength, current on-off ratio (Ion/Ioff), energy-delay product, and power-delay product for logic gates, namely NAND and NOR, are presented. Design rules used for carbon nanotube field-effect transistors (CNTFETs) are compatible with the 45-nm MOSFET technology. The parasitics associated with interconnects are also incorporated in the model. Interconnects can affect the propagation delay in a CNTFET. Smaller length interconnects result in higher cutoff frequency.


Making Common Sense of Vaccines: An Example of Discussing the Recombinant Attenuated Salmonella Vaccine with the Public.
Researchers have iterated that the future of synthetic biology and biotechnology lies in novel consumer applications of crossing biology with engineering. However, if the new biology's future is to be sustainable, early and serious efforts must be made towards social sustainability. Therefore, the crux of new applications of synthetic biology and biotechnology is public understanding and acceptance. The RASVaccine is a novel recombinant design not found in nature that re-engineers a common bacteria (Salmonella) to produce a strong immune response in humans. Synthesis of the RASVaccine has the potential to improve public health as an inexpensive, non-injectable product. But how can scientists move forward to create a dialogue of creating a 'common sense' of this new technology in order to promote social sustainability? This paper delves into public issues raised around these novel technologies and uses the RASVaccine as an example of meeting the public with a common sense of its possibilities and limitations.