Adaptations in Basal and Hypothalamic-Pituitary-Adrenal-Activated Deoxycorticosterone Responses Following Ethanol Self-administration in Cynomolgus Monkeys.
Acute ethanol activates the hypothalamic-pituitary-adrenal (HPA) axis, while long-term exposure results in a blunted neuroendocrine state, particularly with regards to the primary endpoint, cortisol, the primary glucocorticoid produced in the adrenal cortex. However, it is unknown if this dampened neuroendocrine status also influences other adrenocortical steroids. Plasma concentration of the mineralocorticoid and neuroactive steroid precursor deoxycorticosterone (DOC) is altered by pharmacological challenges of the HPA axis in cynomolgus monkeys. The present study investigated HPA axis regulation of circulating DOC concentration over the course of ethanol (4% w/v) induction and self-administration in non-human primates (Macaca fasciculata, n = 10). Plasma DOC, measured by radioimmunoassay, was compared at baseline (ethanol naïve), during schedule-induced polydipsia, and following 6-months of 22 h/day access to ethanol and water. The schedule induction of ethanol drinking did not alter basal DOC levels but selectively dampened the DOC response to pharmacological challenges aimed at the anterior pituitary (ovine corticotrophin-releasing hormone) and adrenal gland (post-dexamethasone adrenocorticotropin hormone), while pharmacological inhibition of central opioid receptors with naloxone greatly enhanced the DOC response during induction. Following 6 months of ethanol self-administration, basal DOC levels were increased more than twofold, while responses to each of the challenges normalized somewhat but remained significantly different than baseline. These data show that HPA axis modulation of the neuroactive steroid precursor DOC is markedly altered by the schedule induction of ethanol drinking and long-term voluntary ethanol self-administration. The consequences of chronic ethanol consumption on HPA axis regulation of DOC point toward allostatic modification of hypothalamic and adrenal function.


Testosterone improves antidepressant-emergent loss of libido in women: findings from a randomized, double-blind, placebo-controlled trial.
Female sexual dysfunction is a side effect of selective serotonin reuptake inhibitor (SSRI)/serotonin noradrenalin reuptake inhibitor (SNRI) therapy. The aim of this study is to investigate the efficacy of transdermal testosterone (TT) as a treatment for SSRI/SNRI-emergent loss of libido. This was a double-blind, randomized, placebo-controlled study. Forty-four women, aged 35-55 years, on a stable dose of SSRI or SNRI with treatment-emergent loss of libido were randomly allocated to treatment with a TT patch delivering 300 mcg of testosterone/day or an identical placebo patch (Pl) for 12 weeks. The primary outcome measure was the change in the Sabbatsberg Sexual Self-rating Scale (SSS) total score over 12 weeks. The 4-week frequency of Satisfactory Sexual Events (SSEs) and the Female Sexual Distress Scale-Revised (FSDS-R) were also measured. At baseline, there were no differences between the treatment groups. At week 12, the change in the SSS score did not differ between the two groups. The increase in the 4-week frequency of SSEs was significantly greater for the TT group than for the Pl group (an increase of 2.3 events vs. 0.1, P = 0.02). The between-group difference in the change in the FSDS-R score approached statistical significance (P = 0.06). The mean total serum testosterone level at 12 weeks in the TT group was 2.1 nmol/L. No women withdrew because of androgenic adverse events. TT therapy resulted in a significant increase in the number of SSEs compared with Pl therapy in women with SSRI/SNRI-emergent loss of libido. The lack of improvement in the SSS total score may reflect lack of sensitivity of this instrument for the measurement of change in sexual function. This provides the first evidence that TT therapy may be a treatment option for women with SSRI/SNRI-emergent loss of libido who need to remain on their antidepressant therapy.


CD1c bypasses lysosomes to present a lipopeptide antigen with 12 amino acids.
The recent discovery of dideoxymycobactin (DDM) as a ligand for CD1a demonstrates how a nonribosomal lipopeptide antigen is presented to T cells. DDM contains an unusual acylation motif and a peptide sequence present only in mycobacteria, but its discovery raises the possibility that ribosomally produced viral or mammalian proteins that commonly undergo lipidation might also function as antigens. To test this, we measured T cell responses to synthetic acylpeptides that mimic lipoproteins produced by cells and viruses. CD1c presented an N-acyl glycine dodecamer peptide (lipo-12) to human T cells, and the response was specific for the acyl linkage as well as the peptide length and sequence. Thus, CD1c represents the second member of the CD1 family to present lipopeptides. lipo-12 was efficiently recognized when presented by intact cells, and unlike DDM, it was inactivated by proteases and augmented by protease inhibitors. Although lysosomes often promote antigen presentation by CD1, rerouting CD1c to lysosomes by mutating CD1 tail sequences caused reduction in lipo-12 presentation. Thus, although certain antigens require antigen processing in lysosomes, others are destroyed there, providing a hypothesis for the evolutionary conservation of large CD1 families containing isoforms that survey early endosomal pathways.


Attentional Selection in a Cocktail Party Environment Can Be Decoded from Single-Trial EEG.
How humans solve the cocktail party problem remains unknown. However, progress has been made recently thanks to the realization that cortical activity tracks the amplitude envelope of speech. This has led to the development of regression methods for studying the neurophysiology of continuous speech. One such method, known as stimulus-reconstruction, has been successfully utilized with cortical surface recordings and magnetoencephalography (MEG). However, the former is invasive and gives a relatively restricted view of processing along the auditory hierarchy, whereas the latter is expensive and rare. Thus it would be extremely useful for research in many populations if stimulus-reconstruction was effective using electroencephalography (EEG), a widely available and inexpensive technology. Here we show that single-trial (≈60 s) unaveraged EEG data can be decoded to determine attentional selection in a naturalistic multispeaker environment. Furthermore, we show a significant correlation between our EEG-based measure of attention and performance on a high-level attention task. In addition, by attempting to decode attention at individual latencies, we identify neural processing at ∼200 ms as being critical for solving the cocktail party problem. These findings open up new avenues for studying the ongoing dynamics of cognition using EEG and for developing effective and natural brain-computer interfaces.


Sensitivity enhancement of surface plasmon resonance biosensing of small molecules.
Surface plasmon resonance (SPR) biosensor formats using gold nanoparticle or protein signal amplification for the sensitive assay of small molecules were developed using progesterone as a model compound. Progesterone was immobilized to a dextran surface in the Biacore biosensor through in situ covalent immobilization using an oligoethylene glycol linker attached to the 4 position of the steroid. This surface produced stable antibody binding for in excess of 1100 assay cycles. Using this surface, assays were developed for progesterone using 10- and 20-nm gold-streptavidin labels attached to biotinylated monoclonal antibody in both label prebinding and sequential binding formats. Prelabeling formats gave no signal enhancement but produced assays with limits of detection of 143 pg/ml, compared with approximately 1 ng/ml in previous studies. Sequential binding formats gave signal enhancements of 2.2-fold over the monoclonal antibody and a limit of detection of 23.1 pg/ml. It was found that secondary antibody labeling gave 8.1-fold signal enhancements and a limit of detection of 20.1 pg/ml, whereas use of secondary antibody-25 nm gold complexes provided more signal enhancement (13-fold) and a further improvement in limit of detection of 8.6 pg/ml.


A Case of Malignant Melanoma with In-Transit Metastasis That Responded to Intravenous Infusion of Interferon-β.
A 77-year-old man with a history of surgical resection of malignant melanoma involving the fifth toe of his left foot 14 years ago presented at the Kariya Toyota General Hospital with a 3-month history of skin ulcer at the same site and red nodules on the lower left leg. Malignant melanoma was suspected, and the patient was referred to our department. On examination, a skin ulcer measuring 25 × 20 mm was observed at the amputation site on the left foot. In addition, multiple red nodules were observed on the lower left leg. Skin biopsies of the ulcer and nodules revealed recurrent malignant melanoma with in-transit metastasis. Two weeks later, he developed acute myocardial infarction and was hospitalized at the Kariya Toyota General Hospital. One month later, the myocardial infarction ameliorated, and he was transferred to our department. As the myocardial infarction had decreased the patient's tolerance to surgery, interferon-β was administered by intravenous infusion. The skin ulcer and red nodules on the lower left leg disappeared 26 weeks after infusion had been initiated. The patient's progress has been satisfactory, with no evidence of recurrence or metastasis at 1 year and 9 months after the initiation of intravenous infusion.


Chondroprotective effects of zoledronic acid on articular cartilage in dogs with experimentally induced osteoarthritis.
To assess effects of zoledronic acid on biomarkers, radiographic scores, and gross articular cartilage changes in dogs with induced osteoarthritis. 21 purpose-bred hound-type dogs. The left stifle joint of each dog was examined arthroscopically to determine initial articular cartilage status, which was followed by cranial cruciate ligament (CrCL) transection to induce osteoarthritis. Dogs were assigned to 3 groups (control group, low dose [10 μg of zoledronic acid/kg], or high dose [25 μg of zoledronic acid/kg). Treatments were administered SC every 3 months for 1 year beginning the day after CrCL transection. Serum and synovial fluid samples and radiographs were obtained 0, 1, 3, 6, 9, and 12 months after transection. At 12 months, each joint was scored for cartilage defects. Serum and synovial fluid biomarkers of bone and cartilage turnover (bone-specific alkaline phosphatase, type I and II collagen, carboxy-propeptide of type II collagen, and chondroitin sulfate 846) were analyzed with ELISAs. The high-dose group had fewer total articular defects and lower severity scores in CrCL-transected stifle joints than did the control group. In addition, the high-dose group had significantly less change in collagenase cleavage of type I or II collagen in the synovial fluid at 1 and 3 months after CrCL transection than did the control group and also had greater changes in bone-specific alkaline phosphatase in synovial fluid at 3 months after CrCL transection than did the control group. Zoledronic acid had a chondroprotective effect in dogs with a transected CrCL.


Phosphatidylinositol (3,4,5)-trisphosphate specifically interacts with the phox homology domain of phospholipase D1 and stimulates its activity.
Phospholipase D (PLD), which catalyzes the hydrolysis of phosphatidylcholine to phosphatidic acid and choline, plays key roles in cellular signal transduction by mediating extracellular stimuli including hormones, growth factors, neurotransmitters, cytokines and extracellular matrix molecules. The molecular mechanisms by which domains regulate the activity of PLD--especially the phox homology (PX) domain--have not been fully elucidated. In this study, we have examined the properties of the PX domains of PLD1 and PLD2 in terms of phosphoinositide binding and PLD activity regulation. Interestingly, the PX domain of PLD1, but not that of PLD2, was found to specifically interact with phosphatidylinositol (3,4,5)-trisphosphate (PtdIns(3,4,5)P3). We found that mutation of the conserved arginine at position 179 of the PLD1 PX domain to lysine or to alanine (R179A or R179K, respectively) disrupts PtdIns(3,4,5)P3 binding. In NIH-3T3 cells, the EGFP-PLD1 PX wild-type domain, but not the two mutants, localized to the plasma membrane after 5-minute treatment with platelet-derived growth factor (PDGF). The enzymatic activity of PLD1 was stimulated by adding PtdIns(3,4,5)P3 in vitro. Treatment with PDGF resulted in the significant increase of PLD1 activity and phosphorylation of the downstream extracellular signal-regulated kinases (ERKs), which was blocked by pre-treatment of HEK 293 cells with phosphoinositide 3-kinase (PI3K) inhibitor after the endogenous PLD2 had been depleted by siRNA specific for PLD2. Nevertheless, both PLD1 mutants (which cannot interact with PtdIns(3,4,5)P3) did not respond to treatment with PDGF. Moreover, PLD1 was activated in HepG2 cells stably expressing the Y40/51 mutant of PDGF receptor that is required for the binding with PI3K. Our results suggest that the PLD1 PX domain enables PLD1 to mediate signal transduction via ERK1/2 by providing a direct binding site for PtdIns(3,4,5)P3 and by activating PLD1.


Marked reduction of alcohol dehydrogenase in keratoconus corneal fibroblasts.
To identify differentially expressed genes in keratoconus (KC) corneal fibroblasts. Stromal keratocytes (having a fibroblast morphology) from KC keratoplasty specimens and eye bank donor corneas were isolated and expanded using a serum containing medium. RNA was isolated from three KC fibroblast cultures and five eye bank donor cornea fibroblast cultures. The targets from the cultured fibroblasts were hybridized to the Affymetrix U133 Plus 2.0 microarrays. Western blot analyses of cell lysates were performed to examine protein levels of interest in the two groups. Protein levels of select differentially expressed genes were further examined by immunohistochemistry. Keratocyte staining of archived KC keratoplasty specimens were graded using a 0 to 3+ scale and compared to five archived whole globes having normal corneas as well as to 10 Fuchs' dystrophy keratoplasty specimens. Microarray analysis revealed up to a 212 fold reduction in the mRNA levels of alcohol dehydrogenase (class 1) beta polypeptide (ADH1B) in KC fibroblasts (p=0.04). Decreased alcohol dehydrogenase in KC fibroblasts was confirmed by western blot analysis of early passage primary keratocyte cell lysates. Immunohistochemistry using a monoclonal mouse immunoglobulin G (IgG) against human liver alcohol dehydrogenase revealed a dramatic difference in protein staining in the keratocytes of the KC group compared to the normal cornea group. Immunohistochemistry also showed decreased immunostaining against alcohol dehydrogenase in the KC stromal sections compared to those obtained from Fuchs' endothelial corneal dystrophy samples. Decreased alcohol dehydrogenase in KC corneal fibroblasts represents a strong marker and possible mediator of keratoconus.


Posttranslational processing of an Epstein-Barr virus-encoded membrane protein expressed in cells transformed by Epstein-Barr virus.
The BamHI Nhet fragment of the B958 strain of Epstein-Barr virus (EBV) encodes a membrane protein (BNLF-1) that is present in cells transformed by EBV. We made a hybrid protein in which a polypeptide sequence from the carboxyl-terminal part of BNLF-1 is fused to Escherichia coli beta-galactosidase. This hybrid protein was used to immunize rabbits, and the resulting antiserum was purified by immunoaffinity chromatography. The antiserum was able to immunoprecipitate BNLF-1 from cell lysates. We found that BNLF-1 is phosphorylated at serines in EBV genome-positive B-cell lines. Pulse-chase analyses with [35S]methionine indicated that BNLF-1 is turned over in lymphoblasts with a half-life of approximately 5 h. Protein immunoblots of EBV genome-positive B-cell lines revealed both a 62,000-molecular-mass band corresponding to BNLF-1 and a myriad of lower-molecular-mass bands. We postulate that these lower-molecular-mass bands are degradation products resulting from the turnover of BNLF-1 in cells. The BNLF-1 gene was expressed in COS cells, and the protein was both phosphorylated and turned over in these cells.


Liposuction-assisted posterior brachioplasty: technical refinements in upper arm contouring.
Brachioplasty has increased 4059 percent from 2000 to 2008 in the United States, with 14,059 upper arm lift procedures performed in 2008. Numerous variations in the evolution of brachioplasty have been described to improve on complications and outcomes. Liposuction-assisted posterior brachioplasty is the next step in the series of refinements in upper arm contouring. The authors present a series of 21 patients who underwent upper arm contouring with liposuction-assisted posterior brachioplasty, and include an operative video detailing the enhancements. After anatomical analysis of the posterior arm, noting skin and fat redundancy, appropriate patients were selected for this procedure. Operative markings, liposuction technique, and the unique excisional technique are presented with intraoperative video footage. Patients tolerate liposuction-assisted posterior brachioplasty very well, with minimal complications and good results. One post-bariatric surgery patient experienced a small wound dehiscence, and one non-bariatric surgery patient developed a slight hypertrophic scar in one arm. No other complications were noted. No revisions were performed. Liposuction-assisted posterior brachioplasty is an efficient and reproducible procedure in selected patients with generalized inferior arm skin and fat redundancy. It simplifies the markings and resection. It provides a safe procedure by preserving lymphatics, blood loss, and nerves. It produces reliable and predictable results with optimal outcomes. This technique offers another refinement in the evolution of upper arm contouring.


Arboviruses and Muscle Disorders: From Disease to Cell Biology.
Infections due to arboviruses (arthropod-borne viruses) have dramatically increased worldwide during the last few years. In humans, symptoms associated with acute infection of most arboviruses are often described as "dengue-like syndrome", including fever, rash, conjunctivitis, arthralgia, and muscular symptoms such as myalgia, myositis, or rhabdomyolysis. In some cases, muscular symptoms may persist over months, especially following flavivirus and alphavirus infections. However, in humans the cellular targets of infection in muscle have been rarely identified. Animal models provide insights to elucidate pathological mechanisms through studying viral tropism, viral-induced inflammation, or potential viral persistence in the muscle compartment. The tropism of arboviruses for muscle cells as well as the viral-induced cytopathic effect and cellular alterations can be confirmed in vitro using cellular models. This review describes the link between muscle alterations and arbovirus infection, and the underlying mechanisms.


Impact of malaria during pregnancy on low birth weight in sub-Saharan Africa.
Malaria during pregnancy can result in low birth weight (LBW), an important risk factor for infant mortality. This article reviews the pathological effects of malaria during pregnancy and the implications for the newborn's development and survival. Empirical data from throughout Africa on associations between placental malaria and birth weight outcome, birth weight outcome and infant mortality, and the rates of LBW in areas with various levels of malaria transmission are evaluated to assess the increased risks of LBW and infant mortality associated with malaria. It is estimated that in areas where malaria is endemic, around 19% of infant LBWs are due to malaria and 6% of infant deaths are due to LBW caused by malaria. These estimates imply that around 100,000 infant deaths each year could be due to LBW caused by malaria during pregnancy in areas of malaria endemicity in Africa.


Macrophage-based anti-cancer therapy: modelling different modes of tumour targeting.
Tumour hypoxia is associated with poor drug delivery and low rates of cell proliferation, factors that limit the efficacy of therapies that target proliferating cells. Since macrophages localise within hypoxic regions, a promising way to target hypoxic tumour cells involves engineering macrophages to express therapeutic genes under hypoxia. In this paper we develop mathematical models to compare the responses of avascular tumour spheroids to two modes of action: either the macrophages deliver an enzyme that activates an externally applied prodrug (bystander model), or they deliver cytotoxic factors directly (local model). The models we develop comprise partial differential equations for a multiphase mixture of tumour cells, macrophages and extracellular fluid, coupled to a moving boundary representing the spheroid surface. Chemical constituents, such as oxygen and drugs, diffuse within the multiphase mixture. Simulations of both models show the spheroid evolving to an equilibrium or to a travelling wave (multiple stable solutions are also possible). We uncover the parameter dependence of the wave speed and steady-state tumour size, and bifurcations between these solution forms. For some parameter sets, adding extra macrophages has a counterintuitive deleterious effect, triggering a bifurcation from bounded to unbounded tumour growth. While these features are common to the bystander and local models, the crucial difference is where cell death occurs. The bystander model is comparable to traditional chemotherapy, with poor targeting of hypoxic tumour cells; however, the local mode of action is more selective for hypoxic regions. We conclude that effective targeting of hypoxic tumour cells may require the use of drugs with limited mobility or whose action does not depend on cell proliferation.


Prognostic implications of coronary artery calcium in the absence of coronary artery luminal narrowing.
Coronary artery calcium (CAC) scoring is a predictor of future adverse clinical events, and a surrogate measure of overall coronary artery plaque burden. Coronary computed tomographic angiography (CCTA) is a contrast-enhanced method that allows for visualization of plaque as well as whether that plaque causes luminal narrowing. To date, the prognosis of individuals with CAC but without stenosis has not been reported. We explored the prevalence of CAC>0 and its prognostic utility for future mortality for patients without luminal narrowing by CCTA. From 17 sites in 9 countries, we identified patients without known coronary artery disease, who underwent CAC scoring and CCTA, and were followed for >3 years. CCTA was graded for % stenosis according to a modified American Heart Association 16-segment model. We calculated hazard ratios (HR) with 95% confidence intervals (95% CI) for incident mortality and compared risk of death for patients as a function of presence or absence of CAC and presence or absence of luminal narrowing by CCTA. Among 6656 patients who underwent CCTA and CAC scoring, 399 patients (6.0%) had no coronary luminal narrowing but CAC>0. During a median follow-up of 5.1 years (IQR: 3.9-5.9 years), 456 deaths occurred. Compared to individuals without luminal narrowing or CAC, individuals without luminal narrowing but CAC>0 were older, more likely to be male and had higher rates of diabetes, hypertension, and dyslipidemia. Individuals without luminal narrowing but CAC experienced a 2-fold increased risk of mortality, with increasing risk of mortality with higher CAC score. Following adjustment, incident death persisted (HR, 1.8; 95% CI, 1.1-2.9, p = 0.02) among patients without luminal narrowing but with CAC>0 compared with patients whose CACS = 0. Individuals without luminal narrowing but CAC ≥100 had mortality risks similar to individuals with non-obstructive CAD (0 < stenosis<50%) by CCTA [HR 2.5 (95% CI 1.3-4.9) and 2.2 (95% CI 1.6-3.0), respectively]. Patients without luminal narrowing but with CAC experience greater risk of 5-year mortality. Patients with CAC score ≥100 and no coronary luminal narrowing experience death rates similar to those with non-obstructive CAD.


Exploring assessment of medical students' competencies in pain medicine-A review.
Considering the continuing high prevalence and public health burden of pain, it is critical that medical students are equipped with competencies in the field of pain medicine. Robust assessment of student expertise is integral for effective implementation of competency-based medical education. The aim of this review was to describe the literature regarding methods for assessing pain medicine competencies in medical students. PubMed, Medline, EMBASE, ERIC, and Google Scholar, and BEME data bases were searched for empirical studies primarily focusing on assessment of any domain of pain medicine competencies in medical students published between January 1997 and December 2016. A total of 41 studies met the inclusion criteria. Most assessments were performed for low-stakes summative purposes and did not reflect contemporary theories of assessment. Assessments were predominantly undertaken using written tests or clinical simulation methods. The most common pain medicine education topics assessed were pain pharmacology and the management of cancer and low-back pain. Most studies focussed on assessment of cognitive levels of learning as opposed to more challenging domains of demonstrating skills and attitudes or developing and implementing pain management plans. This review highlights the need for more robust assessment tools that effectively measure the abilities of medical students to integrate pain-related competencies into clinical practice. A Pain Medicine Assessment Framework has been developed to encourage systematic planning of pain medicine assessment at medical schools internationally and to promote continuous multidimensional assessments in a variety of clinical contexts based on well-defined pain medicine competencies.


cDNA cloning of a wounding-inducible gene encoding a plastid omega-3 fatty acid desaturase from tobacco.
A cDNA encoding the plastid omega-3 fatty acid desaturase was isolated from a tobacco (Nicotiana tabacum cv. SR1) leaf cDNA library. The amino terminal extension of the deduced amino acid sequence of this clone had a characteristic feature of the transit peptides of plastid-destined proteins. Northern analysis indicated that the mRNA corresponding to this cDNA was present in leaves, but was not detected in roots. Responses to wounding of the plastid and microsome omega-3 desaturase genes were investigated in tobacco leaves. The mRNA level of the plastid omega-3 desaturase gene increased to about 2-fold that of unwounded controls at 12 h after a wounding treatment. On the other hand, the mRNA level of the microsome omega-3 desaturase gene remained constant in the wounded leaves. Linolenic acid contents of major leaf polar lipids increased by wounding. These results indicate that wounding enhances the accumulation of the plastid omega-3 desaturase mRNA, and increases the conversion of linoleic acid to linolenic acid in leaf polar lipids.


Analysis of astigmatic focusing and push-pull tracking error signals in magnetooptical disk systems.
Methods of geometrical optics are used in deriving a simple expression for the focus error signal in terms of the parameters of an astigmatic system. The extent of validity of this result is then examined by comparison with diffraction calculations. Diffraction analysis also permits the study of push-pull tracking error signal from pregrooved disk surfaces.


Quantitative Frequency-Domain Passive Cavitation Imaging.
Passive cavitation detection has been an instrumental technique for measuring cavitation dynamics, elucidating concomitant bioeffects, and guiding ultrasound therapies. Recently, techniques have been developed to create images of cavitation activity to provide investigators with a more complete set of information. These techniques use arrays to record and subsequently beamform received cavitation emissions, rather than processing emissions received on a single-element transducer. In this paper, the methods for performing frequency-domain delay, sum, and integrate passive imaging are outlined. The method can be applied to any passively acquired acoustic scattering or emissions, including cavitation emissions. To compare data across different systems, techniques for normalizing Fourier transformed data and converting the data to the acoustic energy received by the array are described. A discussion of hardware requirements and alternative imaging approaches is additionally outlined. Examples are provided in MATLAB.


SIRT2 regulates NF-κB dependent gene expression through deacetylation of p65 Lys310.
NF-κB regulates the expression of a large number of target genes involved in the immune and inflammatory response, apoptosis, cell proliferation, differentiation and survival. In this study, we identified SIRT2 as a deacetylase of the transcription factor p65. SIRT2 is a member of the family of sirtuins, which are NAD(+)-dependent deacetylases involved in several cellular processes. SIRT2 interacts with p65 in the cytoplasm and deacetylates p65 in vitro and in vivo at Lys310. Moreover, p65 is hyperacetylated at Lys310 in Sirt2(-/-) cells after TNFα stimulation, which results in the increase in expression of a subset of p65 acetylation-dependent target genes. Our work provides evidence that p65 is deacetylated by SIRT2 in the cytoplasm to regulate the expression of specific NF-κB-dependent genes.


Mycoplasma inhibition of phytohemagglutinin stimulation of lymphocytes.
Goat lymphocytes were cultured in vitro with phytohemagglutinin and nonviable mycoplasmas. Addition of the mycoplasmas, even as late as 45 hours after adding phytohemagglutinin, completely inhibited the increase in synthesis of DNA and RNA normally induced in lymphocytes by the mitogen. The suppression of synthesis did not result from killing of the cells by the mycoplasmas, combination of the organisms with phytohemagglutinin, or competition for combining sites on the cell surface, which indicates that some other mechanism of inhibition was operative. A similar depression of response to phytohemagglutinin in lymphocytes in culture has been observed in human diseases associated with an immune defect. The present demonstration that at least certain mycoplasmas can profoundly affect lymphocyte function in vitro suggests that thay may alter the immune response in vivo.


Long-term growth comparison studies of FBS and FBS alternatives in six head and neck cell lines.
Fetal bovine serum (FBS) is depended upon by investigators as an indispensable supplement in cell and tissue culture systems. Due to increased demand and limited availability, the price of FBS has increased by greater than 300% in the past few years. In addition, there are ethical and scientific controversies about the collection and use of FBS in culture systems. In response to the shortage of FBS, many FBS alternative serum products have been developed. Although many have claimed comparable performance to FBS, their support of long-term cell growth and effects on cell phenotype have not been revealed. In this study, we examined the performances of six bovine calf serum-based FBS alternatives in six head and neck cell lines and compared them with FBS. The results indicate that some of these sera had growth promoting capabilities comparable or superior to that of FBS. Additionally, these alternative sera supported long-term (30 passages) growth of tested cells and exhibited plating efficiencies comparable to that of FBS. Cells cultured in alternative sera also exhibited comparable anchorage-independent growth and similar drug inhibition responses in FBS. Still, caution should be taken in choosing suitable sera given that changes in cell morphology and variations in chemotactic responses were noted for cells maintained in certain sera. These FBS alternatives are more readily available, cost less, and are associated with less ethical concerns, thus making them attractive alternatives to FBS in cell culture systems.


Work characteristics and upper extremity disorders in female dental health workers.
Many dental health workers suffer from musculoskeletal disorders in the upper extremities. In addition to ergonomic factors, psychosocial work characteristics have been linked to musculoskeletal disorders. The present cross-sectional study aimed at investigating how musculoskeletal disorders in the upper extremities (UED) and occupational position are related to work characteristics and general health problems in female dental health workers. Questionnaire data from dentists, dental hygienists and dental nurses (N=945) showed that 81% reported UED. Multivariate analysis of variance showed that dentists reported the highest levels of physical load and fatigue whereas dental nurses reported the lowest levels of influence at work. Irrespective of position, those with UED considered their physical and psychosocial work environment and their own health to be significantly poorer than did those without UED. A hierarchical multiple regression showed that the physical load of dentistry was most strongly related to UED. Despite improvements to the ergonomics and physical work environment of dentistry, it is concluded that female dental health workers are still at high risk of developing UED.


Double reduction of plastoquinone to plastoquinol in photosystem 1.
In Photosystem 1 (PS1), phylloquinone (PhQ) acts as a secondary electron acceptor from chlorophyll ec(3) and also as an electron donor to the iron-sulfur cluster F(X). PS1 possesses two virtually equivalent branches of electron transfer (ET) cofactors from P(700) to F(X), and the lifetime of the semiquinone intermediate displays biphasic kinetics, reflecting ET along the two different branches. PhQ in PS1 serves only as an intermediate in ET and is not normally fully reduced to the quinol form. This is in contrast to PS2, in which plastoquinone (PQ) is doubly reduced to plastoquinol (PQH(2)) as the terminal electron acceptor. We purified PS1 particles from the menD1 mutant of Chlamydomonas reinhardtii that cannot synthesize PhQ, resulting in replacement of PhQ by PQ in the quinone-binding pocket. The magnitude of the stable flash-induced P(700)(+) signal of menD1 PS1, but not wild-type PS1, decreased during a train of laser flashes, as it was replaced by a ~30 ns back-reaction from the preceding radical pair (P(700)(+)A(0)(-)). We show that this process of photoinactivation is due to double reduction of PQ in the menD1 PS1 and have characterized the process. It is accelerated at lower pH, consistent with a rate-limiting protonation step. Moreover, a point mutation (PsaA-L722T) in the PhQ(A) site that accelerates ET to F(X) ~2-fold, likely by weakening the sole H-bond to PhQ(A), also accelerates the photoinactivation process. The addition of exogenous PhQ can restore activity to photoinactivated PS1 and confer resistance to further photoinactivation. This process also occurs with PS1 purified from the menB PhQ biosynthesis mutant of Synechocystis PCC 6803, demonstrating that it is a general phenomenon in both prokaryotic and eukaryotic PS1.


Endometrial alterations in endometriosis: a systematic review of putative biomarkers.
Endometriosis is usually diagnosed by an invasive procedure such as a laparoscopy. Great interest therefore lies in the potential to identify biomarkers which may be surrogates of disease presence or activity, especially relating to the effects of therapy. We have reviewed the existing literature on endometrial differences in women with endometriosis, and assess their potential use as disease biomarkers. We used QUADAS (Quality Assessment of Diagnostic Accuracy Studies) criteria to conduct a systematic review of published papers over the past 25 years on the subject of endometrial differences in endometriosis. We searched for all studies assessing differences between eutopic endometrium of women with and without endometriosis. We identified 182 relevant articles that are summarized in the review. These studies assess over 200 potential biomarkers, including hormones and their receptors (n = 29), cytokines (n = 25), factors identified using proteomics (n = 8) and histological analysis (n = 10) of endometrial tissue. Sensitivity and specificity were reported or could be calculated for only 32 articles, and ranged from 0 to 100%. Of the nine highest quality studies, six identified putative biomarkers related to nerve fibre growth or cell cycle control, highlighting these areas as promising candidates for future biomarker research. This systematic review identified several reports of endometrial differences which have the potential to be biomarkers of endometriosis. However, larger studies in well-defined populations are clearly required to determine their true usefulness.


Regional and social differences in body mass index, and the prevalence of overweight and obesity among 18 year old men in Austria between the years 1985 and 2000.
The BMI data of four birth cohorts of totally 180,716 male 18 year old Austrian conscripts were documented in 5 year intervals starting 1985 and ending 2000 in order to analyze regional and social variety and a time trend of stature, body weight and BMI as well as the prevalence rates of overweight (defined as a BMI > 25.00) and obesity (BMI > 30.00). At first a marked time trend in BMI and overweight/obesity prevalence rates was found. Over the 15 years of investigation BMI increased significantly and the variation of BMI distribution increased too. The impact of educational level on BMI and the prevalence of overweight and obesity was also statistically significant. With increasing educational level BMI and the prevalence rates of overweight and obesity decreased significantly. Furthermore a significant regional variety in BMI and the prevalence rates of overweight and obesity were found. BMI and the prevalence rates of obesity and overweight decreased significantly from the Eastern part of Austria to the Western part. Both observations, e.g. the social and regional variety of obesity and overweight prevalence, were true of all 4 birth cohorts. From these results we can conclude that obesity and overweight represent an increasing health problem among young Austrian males. This is especially true of young men of low social status living in the eastern part of Austria.


Late Systolic Myocardial Loading Is Associated With Left Atrial Dysfunction in Hypertension.
Late systolic load has been shown to cause diastolic dysfunction in animal models. Although the systolic loading sequence of the ventricular myocardium likely affects its coupling with the left atrium (LA), this issue has not been investigated in humans. We aimed to assess the relationship between the myocardial loading sequence and LA function in human hypertension. We studied 260 subjects with hypertension and 19 normotensive age- and sex-matched controls. Time-resolved central pressure and left ventricular geometry were measured with carotid tonometry and cardiac magnetic resonance imaging, respectively, for computation of time-resolved ejection-phase myocardial wall stress (MWS). The ratio of late/early ejection-phase MWS time integrals was computed as an index of late systolic myocardial load. Atrial mechanics were measured with cine-steady-state free-precession magnetic resonance imaging using feature-tracking algorithms. Compared with normotensive controls, hypertensive participants demonstrated increased late/early ejection-phase MWS and reduced LA function. Greater levels of late/early ejection-phase MWS were associated with reduced LA conduit, reservoir, and booster pump LA function. In models that included early and late ejection-phase MWS as independent correlates of LA function, late systolic MWS was associated with lower, whereas early systolic MWS was associated with greater LA function, indicating an effect of the relative loading sequence (late versus early MWS) on LA function. These relationships persisted after adjustment for multiple potential confounders. A myocardial loading sequence characterized by prominent late systolic MWS was independently associated with atrial dysfunction. In the context of available experimental data, our findings support the deleterious effects of late systolic loading on ventricular-atrial coupling.


Revisiting Lynam's notion of the "fledgling psychopath": are HIA-CP children truly psychopathic-like?
In his developmental model of emerging psychopathy, Lynam proposed that the "fledgling psychopath" is most likely to be located within a subgroup of children elevated in both hyperactivity/inattention/impulsivity (HIA) and conduct problems (CP). This approach has garnered some empirical support. However, the extent to which Lynam's model captures children who resemble psychopathy with regard to the core affective and interpersonal features remains unclear. In the present study, we investigated this issue within a large community sample of youth (N = 617). Four groups (non-HIA-CP, HIA-only, CP-only, and HIA-CP), defined on the basis of teacher reports of the Strengths and Difficulties Questionnaire (SDQ), were compared with respect to parent-reported psychopathic-like traits and subjective emotional reactivity in response to unpleasant, emotionally-laden pictures from the International Affective Pictures System (IAPS). Results did not support Lynam's model. HIA-CP children did not appear most psychopathic-like on dimensions of callous-unemotional and narcissistic personality, nor did they report reduced emotional reactivity to the IAPS relative to the other children. Post-hoc regression analyses revealed a significant moderation such that elevated HIA weakened the association between CP and emotional underarousal. Implications of these findings with regard to the development of psychopathy are discussed.


Determinants of participation in worksite health promotion programmes: a systematic review.
The workplace has been identified as a promising setting for health promotion, and many worksite health promotion programmes have been implemented in the past years. Research has mainly focused on the effectiveness of these interventions. For implementation of interventions at a large scale however, information about (determinants of) participation in these programmes is essential. This systematic review investigates initial participation in worksite health promotion programmes, the underlying determinants of participation, and programme characteristics influencing participation levels. Studies on characteristics of participants and non-participants in worksite health promotion programmes aimed at physical activity and/or nutrition published from 1988 to 2007 were identified through a structured search in PubMed and Web of Science. Studies were included if a primary preventive worksite health promotion programme on PA and/or nutrition was described, and if quantitative information was present on determinants of participation. In total, 23 studies were included with 10 studies on educational or counselling programmes, 6 fitness centre interventions, and 7 studies examining determinants of participation in multi-component programmes. Participation levels varied from 10% to 64%, with a median of 33% (95% CI 25-42%). In general, female workers had a higher participation than men (OR = 1.67; 95% CI 1.25-2.27]), but this difference was not observed for interventions consisting of access to fitness centre programmes. For the other demographic, health- and work-related characteristics no consistent effect on participation was found. Pooling of studies showed a higher participation level when an incentive was offered, when the programme consisted of multiple components, or when the programme was aimed at multiple behaviours. In this systematic review, participation levels in health promotion interventions at the workplace were typically below 50%. Few studies evaluated the influence of health, lifestyle and work-related factors on participation, which hampers the insight in the underlying determinants of initial participation in worksite health promotion. Nevertheless, the present review does provide some strategies that can be adopted in order to increase participation levels. In addition, the review highlights that further insight is essential to develop intervention programmes with the ability to reach many employees, including those who need it most and to increase the generalizability across all workers.


Antithrombotic drug use, cerebral microbleeds, and intracerebral hemorrhage: a systematic review of published and unpublished studies.
Cerebral microbleeds (MB) are potential risk factors for intracerebral hemorrhage (ICH), but it is unclear if they are a contraindication to using antithrombotic drugs. Insights could be gained by pooling data on MB frequency stratified by antithrombotic use in cohorts with ICH and ischemic stroke (IS)/transient ischemic attack (TIA). We performed a systematic review of published and unpublished data from cohorts with stroke or TIA to compare the presence of MB in: (1) antithrombotic users vs nonantithrombotic users with ICH; (2) antithrombotic users vs nonusers with IS/TIA; and (3) ICH vs ischemic events stratified by antithrombotic use. We also analyzed published and unpublished follow-up data to determine the risk of ICH in antithrombotic users with MB. In a pooled analysis of 1460 ICH and 3817 IS/TIA, MB were more frequent in ICH vs IS/TIA in all treatment groups, but the excess increased from 2.8 (odds ratio; range, 2.3-3.5) in nonantithrombotic users to 5.7 (range, 3.4-9.7) in antiplatelet users and 8.0 (range, 3.5-17.8) in warfarin users (P difference=0.01). There was also an excess of MB in warfarin users vs nonusers with ICH (OR, 2.7; 95% CI, 1.6-4.4; P<0.001) but none in warfarin users with IS/TIA (OR, 1.3; 95% CI, 0.9-1.7; P=0.33; P difference=0.01). There was a smaller excess of MB in antiplatelet users vs nonusers with ICH (OR, 1.7; 95% CI, 1.3-2.3; P<0.001), but findings were similar for antiplatelet users with IS/TIA (OR, 1.4; 95% CI, 1.2-1.7; P<0.001; P difference=0.25). In pooled follow-up data for 768 antithrombotic users, presence of MB at baseline was associated with a substantially increased risk of subsequent ICH (OR, 12.1; 95% CI, 3.4-42.5; P<0.001). The excess of MB in warfarin users with ICH compared to other groups suggests that MB increase the risk of warfarin-associated ICH. Limited prospective data corroborate these findings, but larger prospective studies are urgently required.


Microvascular Brain Disease Progression and Risk of Stroke: The ARIC Study.
Data on the significance of combined white matter hyperintensities (WMH)/lacunar brain infarcts, and their progression over time for the prediction of stroke are scarce. We studied associations between the progression in combined measures of microvascular brain disease and risk of stroke in the ARIC study (Atherosclerosis Risk in Communities). Prospective analysis of 907 stroke-free ARIC participants who underwent a brain magnetic resonance imaging (MRI) in 1993 to 1995, a second brain MRI in 2004 to 2006, and were subsequently followed for stroke incidence through December 31, 2017 (median [25%-75%] follow-up 12.6 [8.9-13.4] years). A combined measure of microvascular brain disease was defined at each visit and categorized by progression from first to second brain MRI as no progression; mild progression (increase of ≥1 unit in WMH grade or new lacune), and moderate progression (increase of ≥1 unit in WMH grade and new lacune). All definite/probable ischemic or hemorrhagic incident strokes occurring after this second MRI, and through 2017, were included. Associations between microvascular brain disease, progression in the combined measures, and stroke incidence were studied with Cox proportional hazard models, adjusting for age, sex, race, education level, time from first to second MRI, body mass index, smoking, hypertension, diabetes mellitus, and coronary heart disease. At the second brain MRI (mean age 72), the distribution of the combined measure was 37% WMH grade <2 and no lacune; 57% WMH grade ≥2 or lacune; and 6% WMH grade ≥2 and lacune. No progression in the combined measures was observed in 38% of participants, 57% showed mild progression and 5% showed moderate progression. Sixty-four incident strokes occurred during the follow-up period. Compared with no change in the combined measure, moderate progression of microvascular brain disease was significantly associated with higher risk of stroke (adjusted hazard ratio, 3.00 [95% CI, 1.30-6.94]). Progression of microvascular brain disease, manifesting as both new lacunes and increase in WMHs grade, is related to substantial increase in long-term risk of stroke.


Snake venomics of African spitting cobras: toxin composition and assessment of congeneric cross-reactivity of the pan-African EchiTAb-Plus-ICP antivenom by antivenomics and neutralization approaches.
Venomic analysis of the venoms of Naja nigricollis, N. katiensis, N. nubiae, N. mossambica, and N. pallida revealed similar compositional trends. The high content of cytotoxins and PLA(2)s may account for the extensive tissue necrosis characteristic of the envenomings by these species. The high abundance of a type I α-neurotoxin in N. nubiae may be responsible for the high lethal toxicity of this venom (in rodents). The ability of EchiTAb-Plus-ICP antivenom to immunodeplete and neutralize the venoms of African spitting cobras was assessed by antivenomics and neutralization tests. It partially immunodepleted 3FTx and PLA(2)s and completely immunodepleted SVMPs and CRISPs in all venoms. The antivenom neutralized the dermonecrotic and PLA(2) activities of all African Naja venoms, whereas lethality was eliminated in the venoms of N. nigricollis, N. mossambica, and N. pallida but not in those of N. nubiae and N. katiensis. The lack of neutralization of lethality of N. nubiae venom may be of medical relevance only in relatively populous areas of the Saharan region. The impaired activity of EchiTAb-Plus-ICP against N. katiensis may not represent a major concern. This species is sympatric with N. nigricollis in many regions of Africa, although very few bites have been attributed to it.


[BARRIERS ASSOCIATED WITH BREAST SELF-EXAMINATION AMONG WOMEN 18 TO 50 YEARS OLD: A CROSS SECTIONAL STUDY].
breast cancer is the most common female malignancy and is commonly associated with high levels of morbidity and mortality in developing countries. Breast self-examination (BSE) it is a fundamental method for early detection of the disease. The purpose of the study was to determine the prevalence and barriers associated to BSE in women between 18 to 50 years old. a cross sectional study in 627 healthy women between 18 50 years old from Pamplona, Colombia, was performed. Barriers to the practice of BSE (age, marital status, religion, family history for cancer disease, healthcare professionals counseling and social media), were collected by a structured questionnaire. Prevalence was calculated for each factor and associated barriers were established through a multivariate logistic regression models. at the time of the survey, about 72% of women stated they did not practice BSE in the last month and 45% in the last six months. Regression models show that women who belong to the age group between 30 and 39 years old, women who did not have any counseling from a health professional and those who did not report breast cancer in their family history; were associated as barriers for practicing BSE. the BSE in the study population was low. These data could be used to provide educational intervention programs about the importance of early diagnosis of breast cancer.


Limiting dilution analysis of helper T-cell function.
Limiting dilution analysis has been applied to the study of T-cell 'helper' function in vitro. Using the microculture system one can estimate the numbers of (a) 'helper' T cells involved in specific collaboration with B cells and (b) those T cells which are able, on being activated by their specific antigen, to facilitate the response of B cells to another antigen. Such studies have enabled us to demonstrate that: (1) a single 'helper' T cell was able to activate a single B-cell precursor to detectable antibody production; (2) the 'helper' function of primed T cells was radio-resistant; (3) a minimal estimate of 'helper' frequencies could be obtained in defined cell populations; (4) nonspecific facilitation was directed towards virtually all available B cells of a given specificity if these were challenged with their appropriate particulate antigen; (5) the microculture system offers the opportunity to determine whether specific and non-specific T-cell 'helper' effects are a consequence of the activity of one T-cell type or of differenct subpopulations of T cells.


Chemotherapy and radiation for prostate cancer.
Over the past few decades there is growing appreciation for the role of chemotherapy in treatment of prostate cancer. Initial successful phase III randomized trials in castration resistant prostate cancer (CRPC) have led to additional successful trials in earlier presentations of disease. Given the established role of radiation in management of locally advanced disease and demonstrated efficacy of taxanes in treatment of prostate cancer, optimal combination of radiation and chemotherapy in this patient population has garnered increased attention. Successful phase III trials in this space have given additional stimulus to further exploring combination therapy with radiation. New directions include assessment of additional chemotherapeutic agents including cabazitaxel and PARP inhibitors as well as personalization of therapy with use of genomic testing and other emerging markers to guide therapeutic decisions.


Multiplicity of biliary excretion mechanisms for irinotecan, CPT-11, and its metabolites in rats.
We have reported previously that a canalicular multispecific organic anion transporter (cMOAT) is responsible for the biliary excretion of carboxylate forms of irinotecan, 7-ethyl-10-[4-(1-piperidino)-1 piperidino] carbonyloxy camptothecin (CPT-11), its active metabolite SN-38, and glucuronide conjugate (SN38-Glu) and the lactone form of SN38-Glu in rats. In this paper, the multiplicity of biliary excretion mechanisms for these four anionic compounds was investigated using isolated liver bile canalicular membrane vesicles (CMVs) obtained from Sprague Dawley rats. For the carboxylate form of CPT-11 and the lactone and carboxylate forms of SN38-Glu, ATP-dependent uptake consisted of both high- and low-affinity components in CMVs. Mutual inhibition studies with S-(2,4-dinitrophenyl)glutathione, a representative substrate for cMOAT, and the uptake study using CMVs from Eisai hyperbilirubinemic rats revealed that cMOAT is responsible for the biliary excretion of the low-affinity component of the carboxylate form of CPT-11 and the high-affinity component of both the lactone and carboxylate forms of SN38-Glu, whereas the high-affinity component for CPT-11 and the low-affinity component for SN38-Glu, which are expressed in Eisai hyperbilirubinemic rats, are governed by a transporter different from cMOAT. The carboxylate form of SN-38 was found to be transported by cMOAT alone. We conclude that multiple transporters, including cMOAT, are responsible for the biliary excretion of CPT-11 and its metabolites (anionic forms), and the contribution of each transporter differs greatly, depending on the substrates.


Scoring Functions for Protein-Ligand Binding Affinity Prediction using Structure-Based Deep Learning: A Review.
The rapid and accurate in silico prediction of protein-ligand binding free energies or binding affinities has the potential to transform drug discovery. In recent years, there has been a rapid growth of interest in deep learning methods for the prediction of protein-ligand binding affinities based on the structural information of protein-ligand complexes. These structure-based scoring functions often obtain better results than classical scoring functions when applied within their applicability domain. Here we review structure-based scoring functions for binding affinity prediction based on deep learning, focussing on different types of architectures, featurization strategies, data sets, methods for training and evaluation, and the role of explainable artificial intelligence in building useful models for real drug-discovery applications.


Moderate-Load Muscular Endurance Strength Training Did Not Improve Peak Power or Functional Capacity in Older Men and Women.
The present study determined the effects of muscular endurance strength training on maximum strength and power, functional capacity, muscle activation and hypertrophy in older men and women. Eighty-one men and women acted as an intervention group while 22 acted as non-training controls (age range 64-75 y). Intervention training included super-sets (i.e., paired exercises, immediately performing the second exercises following completion of the first) with short rest intervals (30-60 s between sets) at an intensity of 50-60% one-repetition maximum (1-RM) for 15-20 repetitions. Concentric leg press actions measured maximum strength (1-RM) and concentric peak power. Functional capacity was assessed by maximum speed walking tests (i.e., forward walk, backward walk, timed-up-and-go, and stair climb tests). Quadriceps muscle activation was assessed by surface electromyogram and twitch interpolation technique. Vastus lateralis cross-sectional area was measured by panoramic ultrasound. Compared to control, the intervention groups increased maximum strength (1-RM; men: 10 ± 7% vs. 2 ± 3%, women: 14 ± 9% vs. 1 ± 6% both P < 0.01) and vastus lateralis cross-sectional area (men: 6 ± 7% vs. -3 ± 6%, women: 10 ± 10% vs. 0 ± 4% both P < 0.05). But there were no between-group differences in peak power, muscle activation or functional capacity (e.g., stair climb; men: -5 ± 7% vs. -4 ± 3%, women: -5 ± 6% vs. -2 ± 5% both P > 0.05). While benefits occurred during muscular endurance strength training, specific stimuli are probably needed to target all aspects of age-related health.


Expression of the foraging gene in adult Drosophila melanogaster.
The foraging gene in Drosophila melanogaster, which encodes a cGMP-dependent protein kinase, is a highly conserved, complex gene with multiple pleiotropic behavioral and physiological functions in both the larval and adult fly. Adult foraging expression is less well characterized than in the larva. We characterized foraging expression in the brain, gastric system, and reproductive systems using a T2A-Gal4 gene-trap allele. In the brain, foraging expression appears to be restricted to multiple sub-types of glia. This glial-specific cellular localization of foraging was supported by single-cell transcriptomic atlases of the adult brain. foraging is extensively expressed in most cell types in the gastric and reproductive systems. We then mapped multiple cis-regulatory elements responsible for parts of the observed expression patterns by a nested cloned promoter-Gal4 analysis. The mapped cis-regulatory elements were consistently modular when comparing the larval and adult expression patterns. These new data using the T2A-Gal4 gene-trap and cloned foraging promoter fusion GAL4's are discussed with respect to previous work using an anti-FOR antibody, which we show here to be non-specific. Future studies of foraging's function will consider roles for glial subtypes and peripheral tissues (gastric and reproductive systems) in foraging's pleiotropic behavioral and physiological effects.


How does tetraphenylethylene relax from its excited states?
Tetraphenylethylene is a prototypical example of a molecule displaying aggregation-induced emission. Despite many studies on the optical properties of TPE and its derivatives, the origin of the non-emissive behavior in the gas phase or in dilute solutions has yet to be unravelled. Here, we identify the ultrafast deactivation mechanisms responsible for the fluorescence quenching in isolated TPE.


Shared medical appointments for patients with diabetes mellitus: a systematic review.
Shared medical appointments (SMAs) are an increasingly used system-redesign strategy for improving access to and quality of chronic illness care. We conducted a systematic review of the existing literature on SMA interventions for patients with diabetes in order to understand their impact on outcomes. MEDLINE, EMBASE, CINAHL, PsycINFO, and Web of Science from January 1996 through April 2012. PubMed search updated June 2013. English-language peer-reviewed publications of randomized controlled trials (RCTs), nonrandomized cluster controlled trials, controlled before-and-after studies, or interrupted time-series designs conducted among adult patients with diabetes. Two independent reviewers used prespecified criteria to screen titles and abstracts for full text review. Two different reviewers abstracted data and rated study quality and strength of evidence. When possible, we used random-effects models to synthesize the effects quantitatively, reporting by a weighted difference of the means when the same scale was used across studies, and a standardized mean difference when the scales differed. We measured heterogeneity in study effects using Forest Plots, Cochran's Q, and I(2), and explored heterogeneity by using subgroup analyses for categorical variables and meta-regression analyses for continuous or discrete variables. Outcomes not suitable to meta-analysis were summarized qualitatively. Twenty-five articles representing 17 unique studies compared SMA interventions with usual care. Among patients with diabetes, SMAs improved hemoglobin A1c (∆ = -0.55 percentage points [95 % CI, -0.11 to -0.99]); improved systolic blood pressure (∆ = -5.2 mmHg [95 % CI, -3.0 to -7.4]); and did not improve LDL cholesterol (∆ = -6.6 mg/dl [95 % CI, 2.8 to -16.1]). Nonbiophysical outcomes, including economic outcomes, were reported too infrequently to meta-analyze, or to draw conclusions from. The A1c result had significant heterogeneity among studies, likely secondary to the heterogeneity among included SMA interventions. Heterogeneity among the components of diabetes SMAs leads to uncertainty about what makes a particular SMA successful. SMA interventions improve biophysical outcomes among patients with diabetes. There was inadequate literature to determine SMA effects on patient experience, utilization, and costs.


Mutations that alter the timing and pattern of cubitus interruptus gene expression in Drosophila melanogaster.
The cubitus interruptus (ci) gene is a member of the Drosophila segment polarity gene family and encodes a protein with a zinc finger domain homologous to the vertebrate Gli genes and the nematode tra-1 gene. Three classes of existing mutations in the ci locus alter the regulation of ci expression and can be used to examine ci function during development. The first class of ci mutations causes interruptions in wing veins four and five due to inappropriate expression of the ci product in the posterior compartment of imaginal discs. The second class of mutations eliminates ci protein early in embryogenesis and causes the deletion of structures that are derived from the region including and adjacent to the engrailed expressing cells. The third class of mutations eliminates ci protein later in embryogenesis and blocks the formation of the ventral naked cuticle. The loss of ci expression at these two different stages in embryonic development correlates with the subsequent elimination of wingless expression. Adults heterozygous for the unique ciCe mutation have deletions between wing veins three and four. A similar wing defect is present in animals mutant for the segment polarity gene fused that encodes a putative serine/threonine kinase. In ciCe/+ and fused mutants, the deletions between wing veins three and four correlate with increased ci protein levels in the anterior compartment. Thus, proper regulation of both the ci mRNA and protein appears to be critical for normal Drosophila development.


Influence of 2 Digital Exercise Modules of a Multimodular System on Balance and Leg Strength Under Consideration of Use Adherence: Prospective Cohort Study.
To empower healthy aging, digital solutions embed multiple modules for physical activity, cognitive health promotion, and social engagement. Integrating new empowering technologies such as digital exercise monitoring requires assessment measures and analysis procedures, considering variable compliance of users with different modules. This study aims to assess the influence of a tablet-based and a feedback system-based exercise module on balance and leg strength by considering use adherence instead of the use of the entire multimodular system. In the prospective cohort study within the fit4AAL project, 83 users (n=67, 81% women; n=16, 19% men; mean age 66.2, SD 2.3 years) used the 2 digital exercise modules of a multimodular physical activity promotion system for >18 weeks. A data-driven clustering method based on the average use frequency of the exercise modules determined the number of user types that met the World Health Organization-recommended training frequency of at least twice per week. On the basis of this use adherence, statistical analysis was performed with features of functional performance tests (unipedal stance, 30-second chair rise, Y-balance, and hurdle step tests). The tests were conducted 6 months before the intervention, immediately before the intervention, and after the intervention, comparing the baseline phase with the 3 feedback use groups of the study (using only the tablet, the tablet and the feedback system, or only the feedback system). Of the 83 users, 43 (52%) met the World Health Organization-recommended frequency of muscle-strengthening activities. Overall, the feedback use groups achieved, on average, more chair rises in 30 seconds than the baseline group (P=.01; moderate effect size of 0.07). Of the 43 users, 26 (60%) additionally used the feedback system-based exercise module. They improved in balance compared with the users using either the tablet or the feedback system (P=.02). In addition, they improved their leg strength within the group (P=.04) and compared with the baseline (P=.01). The additional use of a feedback system showed a tendency to positively maintain and influence the already exceptionally high functional performance of older adults. Considering use adherence in future multimodular system studies is crucial to assess the influence of single and combined use of exercise modules on functional performance.


Universal test-and-treat in Zambian and South African correctional facilities: a multisite prospective cohort study.
Despite the global scale-up of antiretroviral therapy (ART), incarcerated people have not benefited equally from test-and-treat recommendations for HIV. To improve access to ART for incarcerated people with HIV, we introduced a universal test-and-treat (UTT) intervention in correctional facilities in South Africa and Zambia, and aimed to assess UTT feasibility and clinical outcomes. Treatment as Prevention (TasP) was a multisite, mixed methods, implementation research study done at three correctional complexes in South Africa (Johnannesburg and Breede River) and Zambia (Lusaka). Here, we report the clinical outcomes for a prospective cohort of incarcerated individuals who were offered the TasP UTT intervention. Incarcerated individuals were eligible for inclusion if they were aged 18 years or older, with new or previously diagnosed HIV, not yet on ART, and were expected to remain incarcerated for 30 days or longer. To enable the implementation of UTT at the included correctional facilities, we first strengthened on-site HIV service delivery. All participants were offered same-day ART initiation, and had two study-specific follow-up visits scheduled to coincide with routine clinic visits at 6 and 12 months. The main outcomes were ART uptake, time from cohort enrolment to ART initiation, and retention in care and viral suppression at 6 and 12 months. We estimated the association between baseline demographic characteristics and time to ART initiation using Cox proportional hazard models, and, in a post-hoc analysis, we used logistic regression models to assess the association between demographic and clinical variables, including time to ART initiation, and the proportion of participants with a composite poor outcome (defined as viral load >50 copies per mL, or for participants with a missing viral load, lack of retention in care in the on-site ART programme) at 6 months. This study is registered at ClinicalTrials.gov, NCT02946762. Between June 23, 2016, and Dec 31, 2017, we identified 1562 incarcerated people with HIV, of whom 1389 (89%) were screened, 1021 (74%) met eligibility criteria, and 975 (95%) were enrolled and followed up to March 31, 2018. At the end of follow-up, 835 (86%) of 975 participants had started ART. Median time from enrolment to ART initiation was 0 days (IQR 0-8). Of 346 participants who remained incarcerated at 6 months, 327 (95%) were retained in care and 269 (78%) had a documented viral load, of whom 262 (97%) achieved viral suppression (<1000 copies per mL). The mortality rate among the 835 participants who had initiated ART was 1·9 per 100 person-years (95% CI 0·9-3·9). No statistically significant associations were identified between any baseline characteristics and time to ART initiation or composite poor outcome. UTT implementation is feasible in correctional settings, and can achieve levels of same-day ART uptake, retention in care, and viral suppression among incarcerated people with HIV that are comparable to those observed in community settings. UK Department for International Development, Swedish International Development Cooperation Agency, Norwegian Agency for Development Cooperation.


Predicting condom use in adolescents: a test of three socio-cognitive models using a structural equation modeling approach.
The theory of planned behavior (TPB), socio-cognitive model (SCM), and information-motivation-behavioral skills (IMB) model are effective in predicting condom use. However, the adequacy of these three theoretical models in predicting the frequency of condom use (FCU) among young people has not been compared. This cross-sectional study tested the applicability and suitability of these three models in predicting the FCU, and analyzed the relationships among the postulated constructs. Sexually experienced adolescents (n = 410) aged 13-18 completed a survey assessing the TPB, SCM, and IMB model constructs. Participants were students recruited from 18 high schools, randomly selected from the north, south, east, and southeast of Spain. A structural equation modelling (SEM) analysis was applied to test TPB, SCM and IBM and constructs relationships of each model using R. The results of SEM demonstrated that behavioral skills predict behavior via motivation as hypothesized by the IMB model, but not directly via knowledge about condom use and sexually transmitted infections (STIs). Cognitive factors, such knowledge about condom use and STIs as well as condom use self-efficacy, directly predicted the FCU when modeled as per the SCM. According to the TPB, condom use intention was the best predictor of the FCU, and condom use intention was predicted by attitudes toward condom use and subjective norms related to condom use, but perceived control was not directly or indirectly related to the FCU. Based on the data, the TPB becomes the best-fit model for predicting the FCU among young people compared to the SCM and IMB model. From a statistical perspective, the TPB seems to be the most suitable model for predicting the FCU among young people compared to the other models. Overall, key direct predictors of the FCU in adolescents included condom use intention, behavioral skills and cognitive factors, such as STIs knowledge and condom use self-efficacy. The next step should be to test integrative models that include personal, contextual, environmental, and social factors.


Study protocol: safety and efficacy of propranolol in newborns with Retinopathy of Prematurity (PROP-ROP): ISRCTN18523491.
Despite new therapeutic approaches have improved the prognosis of newborns with retinopathy of prematurity (ROP), an unfavourable structural and functional outcome still remains high. There is high pressure to develop new drugs to prevent and treat ROP. There is increasing enthusiasm for anti-VEGF drugs, but angiogenic inhibitors selective for abnormal blood vessels would be considered as an optimal treatment.In an animal experimental model of proliferative retinopathy, we have recently demonstrated that the pharmacological blockade of beta-adrenoreceptors improves retinal neovascularization and blood retinal barrier breakdown consequent to hypoxia. The purpose of this study is to evaluate the propranolol administration in preterm newborns suffering from a precocious phase of ROP in terms of safety and efficacy in counteracting the progression of retinopathy. Preterm newborns (gestational age at birth lower than 32 weeks) with stage 2 ROP (zone II-III without plus) will be randomized, according to their gestational age, to receive propranolol added to standard treatment (treatment adopted by the ETROP Cooperative Group) or standard treatment alone. Propranolol will be administered until retinal vascularization will be completely developed, but not more than 90 days. Forty-four participants will be recruited into the study. To evaluate the safety of propranolol administration, cardiac and respiratory parameters will be continuously monitored. Blood samplings will be performed to check renal, liver and metabolic balance. To evaluate the efficacy of propranolol, the progression of the disease, the number of laser treatments or vitrectomies, the incidence of retinal detachment or blindness, will be evaluated by serial ophthalmologic examinations. Visual function will be evaluated by means of behavioural standardized tests. This pilot study is the first research that explores the possible therapeutic role of beta blockers in ROP. The objective of this research is highly ambitious: to find a treatment simple, inexpensive, well tolerated and with few adverse effects, able to counteract one of the major complications of the prematurity. Any favourable results of this research could open new perspectives and original scenarios about the treatment or the prevention of this and other proliferative retinopathies. Current Controlled Trials ISRCTN18523491; ClinicalTrials.gov Identifier NCT01079715; EudraCT Number 2010-018737-21.


A Co-Expression Network in Hexaploid Wheat Reveals Mostly Balanced Expression and Lack of Significant Gene Loss of Homeologous Meiotic Genes Upon Polyploidization.
Polyploidization has played an important role in plant evolution. However, upon polyploidization, the process of meiosis must adapt to ensure the proper segregation of increased numbers of chromosomes to produce balanced gametes. It has been suggested that meiotic gene (MG) duplicates return to a single copy following whole genome duplication to stabilize the polyploid genome. Therefore, upon the polyploidization of wheat, a hexaploid species with three related (homeologous) genomes, the stabilization process may have involved rapid changes in content and expression of MGs on homeologous chromosomes (homeologs). To examine this hypothesis, sets of candidate MGs were identified in wheat using co-expression network analysis and orthology informed approaches. In total, 130 RNA-Seq samples from a range of tissues including wheat meiotic anthers were used to define co-expressed modules of genes. Three modules were significantly correlated with meiotic tissue samples but not with other tissue types. These modules were enriched for GO terms related to cell cycle, DNA replication, and chromatin modification and contained orthologs of known MGs. Overall, 74.4% of genes within these meiosis-related modules had three homeologous copies which was similar to other tissue-related modules. Amongst wheat MGs identified by orthology, rather than co-expression, the majority (93.7%) were either retained in hexaploid wheat at the same number of copies (78.4%) or increased in copy number (15.3%) compared to ancestral wheat species. Furthermore, genes within meiosis-related modules showed more balanced expression levels between homeologs than genes in non-meiosis-related modules. Taken together, our results do not support extensive gene loss nor changes in homeolog expression of MGs upon wheat polyploidization. The construction of the MG co-expression network allowed identification of hub genes and provided key targets for future studies.


Bloodstream and central line isolates from hematopoietic stem cell transplant recipients: data from a developing country.
Bloodstream infections (BSIs) and central line infections remain among the major causes of morbidity and mortality in transplant recipients because of prolonged neutropenia and mucosal damage. The objective of this study was to determine the frequency and outcome of bacterial and fungal isolates from patients undergoing allogeneic hematopoietic stem cell transplant. This study was conducted at the Aga Khan University and Hospital's bone marrow transplant unit. All patients who underwent an allogeneic stem cell transplant with matched sibling/parent donor were included. The study period ranged from April 2004 to December 2012. Transplantation was performed according to institutional protocols. All patients were admitted in single rooms with positive pressure and high-efficiency particulate air filters. Ciprofloxacin, fluconazole, and valaciclovir were used for standard prophylaxis, which was started at the time of conditioning. All blood cultures were obtained at clinical suspicion of systemic infection, mainly documented as fever (temperature of >38.5°C). BSIs and line infections were defined as isolation of bacterial or fungal pathogen from at least one blood/central line culture. In total, 101 of 108 patients developed febrile neutropenia. In the 101 patients, 245 documented febrile episodes occurred. There were 40 culture-positive episodes and 205 culture-negative episodes. Of these 40 culture-positive episodes, 22 patients had bloodstream isolates and 18 had central line isolates. The median ± standard deviation time of febrile neutropenia was day 7 ± 2 days (range: day -3 to day +13). The most common bloodstream isolate was Escherichia coli (n = 9) followed by Staphylococcus epidermidis (n = 5). One patient developed Fusarium infection. In central line infections, S. epidermidis was the most common organism (n = 8). In 2 patients with central venous catheters, Candida albicans was the isolate. Transplant-related mortality from sepsis occurred in 9.2%. E.coli was mainly responsible for BSI, while gram-positive organisms dominated catheter-related febrile episodes. Transplant-related mortality due to sepsis was 9%.


A very early-branching Staphylococcus aureus lineage lacking the carotenoid pigment staphyloxanthin.
Here we discuss the evolution of the northern Australian Staphylococcus aureus isolate MSHR1132 genome. MSHR1132 belongs to the divergent clonal complex 75 lineage. The average nucleotide divergence between orthologous genes in MSHR1132 and typical S. aureus is approximately sevenfold greater than the maximum divergence observed in this species to date. MSHR1132 has a small accessory genome, which includes the well-characterized genomic islands, νSAα and νSaβ, suggesting that these elements were acquired well before the expansion of the typical S. aureus population. Other mobile elements show mosaic structure (the prophage ϕSa3) or evidence of recent acquisition from a typical S. aureus lineage (SCCmec, ICE6013 and plasmid pMSHR1132). There are two differences in gene repertoire compared with typical S. aureus that may be significant clues as to the genetic basis underlying the successful emergence of S. aureus as a pathogen. First, MSHR1132 lacks the genes for production of staphyloxanthin, the carotenoid pigment that confers upon S. aureus its characteristic golden color and protects against oxidative stress. The lack of pigment was demonstrated in 126 of 126 CC75 isolates. Second, a mobile clustered regularly interspaced short palindromic repeat (CRISPR) element is inserted into orfX of MSHR1132. Although common in other staphylococcal species, these elements are very rare within S. aureus and may impact accessory genome acquisition. The CRISPR spacer sequences reveal a history of attempted invasion by known S. aureus mobile elements. There is a case for the creation of a new taxon to accommodate this and related isolates.


Human parainfluenza virus-3 can be targeted by rapidly ex vivo expanded T lymphocytes.
Human parainfluenza virus-3 (HPIV) is a common cause of respiratory infection in immunocompromised patients and currently has no effective therapies. Virus-specific T-cell therapy has been successful for the treatment or prevention of viral infections in immunocompromised patients but requires determination of T-cell antigens on targeted viruses. HPIV3-specific T cells were expanded from peripheral blood of healthy donors using a rapid generation protocol targeting four HPIV3 proteins. Immunophenotyping was performed by flow cytometry. Viral specificity was determined by interferon (IFN)-γ ELISpot, intracellular cytokine staining and cytokine measurements from culture supernatants by Luminex assay. Cytotoxic activity was tested by 51Cr release and CD107a mobilization assays. Virus-specific T cells targeting six viruses were then produced by rapid protocol, and the phenotype of HPIV3-specific T cells was determined by immunomagnetic sorting for IFN-γ-producing cells. HPIV3-specific T cells were expanded from 13 healthy donors. HPIV3-specific T cells showed a CD4+ predominance (mean CD4:CD8 ratio 2.89) and demonstrated specificity for multiple HPIV3 antigens. The expanded T cells were polyfunctional based on cytokine production but only had a minor cytotoxic component. T cells targeting six viruses in a single product similarly showed HPIV3 specificity, with a predominant effector memory phenotype (CD3+/CD45RA-/CCR7-) in responder cells. HPIV3-specific T cells can be produced using a rapid ex vivo protocol from healthy donors and are predominantly CD4+ T cells with Th1 activity. HPIV3 epitopes can also be successfully targeted alongside multiple other viral epitopes in production of six-virus T cells, without loss of HPIV3 specificity. These products may be clinically beneficial to combat HPIV3 infections by adoptive T-cell therapy in immune-compromised patients.


Parenthood--a contributing factor to childhood obesity.
Prevalence of childhood obesity and its complications have increased world-wide. Parental status may be associated with children's health outcomes including their eating habits, body weight and blood cholesterol. The National Health and Nutrition Examination Survey (NHANES) for the years 1988-1994, provided a unique opportunity for matching parents to children enabling analyses of joint demographics, racial differences and health indicators. Specifically, the NHANES III data, 1988-1994, of 219 households with single-parents and 780 dual-parent households were analyzed as predictors for primary outcome variables of children's Body Mass Index (BMI), dietary nutrient intakes and blood cholesterol. Children of single-parent households were significantly (p < 0.01) more overweight than children of dual-parent households. Total calorie and saturated fatty acid intakes were higher among children of single-parent households than dual-parent households (p < 0.05). On average, Black children were more overweight (p < 0.04) than children of other races. The study results implied a strong relationship between single-parent status and excess weight in children. Further studies are needed to explore the dynamics of single-parent households and its influence on childhood diet and obesity. Parental involvement in the development of school- and community-based obesity prevention programs are suggested for effective health initiatives. Economic constraints and cultural preferences may be communicated directly by family involvement in these much needed public health programs.


Feasibility, Safety, and Periprocedural Complications of Pipeline Embolization for Intracranial Aneurysm Treatment Under Conscious Sedation: University at Buffalo Neurosurgery Experience.
Endovascular Pipeline Embolization Device (PED) placement for intracranial aneurysms is performed under general anesthesia at most centers because of perceived improved image quality and patient safety. To report the feasibility, safety, and outcomes associated with the use of the PED for intracranial aneurysms performed in awake patients after the administration of conscious sedation (CS) and a local anesthetic. Between March 2012 and September 2014, 130 patients with 139 intracranial aneurysms (8 ruptured) were treated with the PED under CS at our institution. Procedure details and time (including duration, radiation exposure, and fluoroscopy) and procedure-related complications were retrospectively reviewed. A total of 155 PED deployment procedures were performed under CS. Treatment was successfully completed in all cases. Anesthesia was converted from CS to general anesthesia during 5 procedures. Mean interval from patient entry at the endovascular suite to procedure initiation was 18 minutes (range, 5 minutes-1 hour 10 minutes). Mean procedure length was 1 hour 25 minutes (range, 30 minutes-3 hours 51 minutes). Mean ± SD values for fluoroscopy time and radiation exposure were 36.17 ± 18.4 minutes and 1367 ± 897 mGy, respectively. The mean amount of contrast material administered was 211.37 ± 83.5 mL. Permanent neurological complications were seen in 4 patients (3%). In our experience, CS for PED placement for intracranial aneurysm treatment is feasible and safe. Procedure and fluoroscopy times and amount of radiation exposure are similar to or less than described in reports of PED placement under general anesthesia. CS allows direct neurological evaluation and earlier detection of and response to intraprocedural complications.


Hepatitis B and C virus infection in the hemodialysis population from three romanian regions.
After 10 years of systematically nationwide applied measures for reduction of infection risk, in this national prospective observational study, we reassessed the prevalence of hepatitis virus infection prevalence and its influence on the outcome of end-stage kidney disease (ESKD) patients treated with hemodialysis. Six-hundred ESKD patients (332 men and 268 women, median age 56 years) treated with chronic HD in seven centers from all the historical regions of Romania have been assigned to this study on 1st of November 2010. The aims of this study were to reevaluate the prevalence of the hepatitis B and C virus infection in a HD population from Romania after 10 years of systematically nationwide applied measures for reduction of infection risk and also to assess the impact of these infections on the prognosis of HD patients. HBsAg was positive in 9.5% (n = 57) of the patients, anti-HCV antibodies were detected in 27.3% (n = 164) and 5% (n = 30) were positive for both HBV and HCV infection. The mortality risk was significantly influenced only by age, the presence of coronary artery disease and the 25 OH vitamin D levels. This study shows that the systematically nationwide applied measures for reduction of infection risk significantly decreased HV infection prevalence in HD patients in Romania. The presence of HV infection did not significantly influence the mortality risk in this population.


Salmonella montevideo infection in sheep and cattle in Scotland, 1970-81.
Outbreaks of abortion associated with infection by Salmonella montevideo have affected sheep in the east, especially the south-east, of Scotland each year since 1972. Disease in the north and north-east was usually less severe. Between 1 January 1970 and 31 December 1981, a total of 67 incidents affecting sheep were reported by veterinary laboratories to the Communicable Diseases (Scotland) Unit, 87% of which presented during the main lambing months of February, March and April. Twenty-one episodes of bovine infection were also recorded over the same period, 17 of which involved single animals only, usually an aborted cow or a scouring calf. Despite intensive investigations, neither the origin nor the mode of spread of S. montevideo infection among sheep and cattle in Scotland have been established with any certainty, although there has been considerable evidence indicating the role of scavenging wild birds, particularly seagulls, as vectors transmitting infection to other farms in the same district. Also largely unexplained are the differences in the epidemiology and clinical pattern of disease in the south-east compared to the north and north-east, while sheep in the west of Scotland have remained virtually unaffected throughout.


Regulation of angiogenic factors in angiotensin II infusion model in association with tubulointerstitial injuries.
Among various angiogenic factors, vascular endothelial growth factor (VEGF), angiopoietin-1 (Ang1), and angiopoietin-2 (Ang2) play crucial roles in regulating angiogenesis and vascular integrity. Infusion of angiotensin-II (ang II) induces hypertension and focal renal tubulointerstitial injuries. In the present study we investigated the renal expression of VEGF, Ang1, Ang2, and corresponding receptors in association with tubulointerstitial lesions in a rat ang II infusion model. Male Sprague-Dawley (SD) rats received an infusion of ang II or norepinephrine (NE) through osmotic minipumps for 14 days. Angiotensin II type 1 (AT1) or type 2 (AT2) receptor antagonist (losartan or PD123319, respectively) or hydralazine was co-administered. Interstitial fibrosis, infiltration of monocyte/macrophage, and peritubular capillary rarefaction induced by ang II was significantly attenuated in the losartan- or PD123319-treated groups. Immunoreactivity of VEGF and Ang1 in cortical tubules was increased by ang II and was attenuated by losartan or PD123319. The increase of VEGF induced by ang II was suppressed by losartan, and the increase of Ang1 induced by ang II was inhibited by PD123319 as detected by immunoblot. The increase of flk-1 and flt-1 (VEGF receptors) and tie-2 (Ang1 receptor) induced by ang II was significantly suppressed by PD123319. These alterations were not observed in hydralazine plus ang II or NE-infused animals. These results demonstrate that an infusion of ang II induced the expression of VEGF mainly through AT1 receptors, and increased the expression of VEGF receptors, tie-2, and Ang1/Ang2 ratio mainly through AT2 receptors. The increase of VEGF/flk-1/flt-1 may be associated with vascular permeability, monocyte/macrophage infiltration, and rarefaction of peritubular capillaries, and the increase of the Ang1/Ang2 ratio may be a compensatory mechanism counteracting the permeability inducing effect of VEGF after ang II infusion.


Effects of B Cell Depletion on Early Mycobacterium tuberculosis Infection in Cynomolgus Macaques.
Although recent studies in mice have shown that components of B cell and humoral immunity can modulate the immune responses against Mycobacterium tuberculosis, the roles of these components in human and nonhuman primate infections are unknown. The cynomolgus macaque (Macaca fascicularis) model of M. tuberculosis infection closely mirrors the infection outcomes and pathology in human tuberculosis (TB). The present study used rituximab, an anti-CD20 antibody, to deplete B cells in M. tuberculosis-infected macaques to examine the contribution of B cells and humoral immunity to the control of TB in nonhuman primates during the acute phase of infection. While there was no difference in the overall pathology, disease profession, and clinical outcome between the rituximab-treated and untreated macaques in acute infection, analyzing individual granulomas revealed that B cell depletion resulted in altered local T cell and cytokine responses, increased bacterial burden, and lower levels of inflammation. There were elevated frequencies of T cells producing interleukin-2 (IL-2), IL-10, and IL-17 and decreased IL-6 and IL-10 levels within granulomas from B cell-depleted animals. The effects of B cell depletion varied among granulomas in an individual animal, as well as among animals, underscoring the previously reported heterogeneity of local immunologic characteristics of tuberculous granulomas in nonhuman primates. Taken together, our data clearly showed that B cells can modulate the local granulomatous response in M. tuberculosis-infected macaques during acute infection. The impact of these alterations on disease progression and outcome in the chronic phase remains to be determined.


Maintenance of Aerobic or Resistance Training After an Exercise Intervention Among Breast Cancer Patients After Neoadjuvant Chemotherapy.
Exercise interventions have been shown to be beneficial for cancer patients regarding various treatment-related side effects and quality of life. For sustainable effects, patients should continue the training. Therefore, we investigated the maintenance of an exercise training in breast cancer patients, reasons for (dis)continuation, and explored possible influencing factors. The investigation is based on a 3-arm randomized intervention trial comparing aerobic and resistance training (19 [4]) during or after neoadjuvant chemotherapy among breast cancer patients. About 2 years after breast surgery, 68 patients (age 52 [11] y) provided information about training continuation, self-reported reasons of (dis)continuation, sociodemographics, employment status, age, and body mass index. Training continuation was investigated with Kaplan-Meier analyses. The intervention was rated as good or very good by 88.1% of participants. Nevertheless, 52.9% discontinued the training directly, but half of them changed to different types of exercise. Reasons for discontinuation included lack of time and long travel distance to the training facility. The median continuation was 19.0 months (Q1, Q3: 5.5, 36.0) with no statistically significant difference between the intervention groups. Younger, better educated, partnered patients tended toward longer training continuation. The majority of patients continued exercising after the end of intervention. However, a nonnegligible number discontinued training immediately or after few months. Practical, social, and financial support for a transition to an adequate training that is affordable and feasible in the patient's daily life might foster training maintenance. Especially patients who are less educated, elderly, or living alone may need more support to continue exercising.


A problem shared is a problem reduced: seeking efficiency in the conservation of felids and primates.
Threats faced by mammalian species can be grouped into one of a handful of categories, such as habitat loss, unsustainable hunting and persecution. Insofar as they face common threats, diverse species may benefit from the same conservation intervention, thereby offering efficiencies in conservation action. We explore this proposition for primates and felids by examining coarse scale overlaps in geographical distributions, using IUCN Red List assessments of the primary threats posed to each species. A global analysis of primates and felids that face common threats reveals the greatest overlap is in Central and South Asia, where up to 14 primates and felids co-occur. More than 80% of the land where at least 1 threatened species of either primate or felid occurs also contains at least one threatened species of the other taxon, yet over 60% of these grid cells containing both threatened primates and felids lie outside Conservation International's hot spots. A review of IUCN Action Plans of the threats to felids and primates strongly supports the hypothesis that they are often the same and occur in the same place. In principle, steps to conserve big cats have the potential to benefit several species of threatened primates, and vice versa.


Escherichia coli DNA polymerase I (Klenow fragment) uses a hydrogen-bonding fork from Arg668 to the primer terminus and incoming deoxynucleotide triphosphate to catalyze DNA replication.
Interactions between the minor groove of the DNA and DNA polymerases appear to play a major role in the catalysis and fidelity of DNA replication. In particular, Arg668 of Escherichia coli DNA polymerase I (Klenow fragment) makes a critical contact with the N-3-position of guanine at the primer terminus. We investigated the interaction between Arg668 and the ring oxygen of the incoming deoxynucleotide triphosphate (dNTP) using a combination of site-specific mutagenesis of the protein and atomic substitution of the DNA and dNTP. Hydrogen bonds from Arg668 were probed with the site-specific mutant R668A. Hydrogen bonds from the DNA were probed with oligodeoxynucleotides containing either guanine or 3-deazaguanine (3DG) at the primer terminus. Hydrogen bonds from the incoming dNTP were probed with (1 'R,3 'R,4 'R)-1-[3-hydroxy-4-(triphosphorylmethyl)cyclopent-1-yl]uracil (dcUTP), an analog of dUTP in which the ring oxygen of the deoxyribose moiety was replaced by a methylene group. We found that the pre-steady-state parameter kpol was decreased 1,600 to 2,000-fold with each of the single substitutions. When the substitutions were combined, there was no additional decrease (R668A and 3DG), a 5-fold decrease (3DG and dcUTP), and a 50-fold decrease (R668A and dcUTP) in kpol. These results are consistent with a hydrogen-bonding fork from Arg668 to the primer terminus and incoming dNTP. These interactions may play an important role in fidelity as well as catalysis of DNA replication.


Thymus-dependent lymphocytes: destruction by lymphocytic choriomeningitis virus.
Selective destruction of small lymphocytes in the thymusdependent areas of lymph nodes and thymocytes was observed in mice infected with lymphocytic choriomeningitis virus. These changes were clearly evident in lymphoid and splenic tissue 3 days after infection and in the thymus by day 7. The destructive changes paralleled growth of the virus in these organs. The findings show that infection with lymphocytic choriomeningitis virus can temporarily cause the equivalent effect of neonatal thymectomy, that is, a "viral thymectomy," which appears to be related to the ability of this virus to cause persistent infection.