Detecting drug interactions from adverse-event reports: interaction between paroxetine and pravastatin increases blood glucose levels.
The lipid-lowering agent pravastatin and the antidepressant paroxetine are among the most widely prescribed drugs in the world. Unexpected interactions between them could have important public health implications. We mined the US Food and Drug Administration's (FDA's) Adverse Event Reporting System (AERS) for side-effect profiles involving glucose homeostasis and found a surprisingly strong signal for comedication with pravastatin and paroxetine. We retrospectively evaluated changes in blood glucose in 104 patients with diabetes and 135 without diabetes who had received comedication with these two drugs, using data in electronic medical record (EMR) systems of three geographically distinct sites. We assessed the mean random blood glucose levels before and after treatment with the drugs. We found that pravastatin and paroxetine, when administered together, had a synergistic effect on blood glucose. The average increase was 19 mg/dl (1.0 mmol/l) overall, and in those with diabetes it was 48 mg/dl (2.7 mmol/l). In contrast, neither drug administered singly was associated with such changes in glucose levels. An increase in glucose levels is not a general effect of combined therapy with selective serotonin reuptake inhibitors (SSRIs) and statins.


Liquid state DNP of water at 9.2 T: an experimental access to saturation.
We have performed liquid state ("Overhauser") Dynamic Nuclear Polarization (DNP) experiments at high magnetic field (9.2 T, corresponding to 260 GHz EPR and 400 MHz (1)H-NMR resonance frequency) on aqueous solutions of (14)N-TEMPOL nitroxide radicals. Integrated signal enhancements exceeding -80 were observed for the water protons at microwave superheated temperatures (160 °C) and still -14 at ambient temperatures (45 °C) relevant to biological applications. Different contributions contributing to the DNP enhancement such as saturation factor, leakage factor and sample temperature under microwave irradiation could be determined independently for a high spin concentration of 1 M, allowing the calculation of the coupling factors as a function of temperature and a quantitative comparison of this parameter with values derived from field dependent relaxation measurements or predictions from MD simulation.


The importance of being expert: the quest for cancer information on the Internet.
To people with the necessary technology the internet can provide vast amounts of health information. However, there are concerns about the quality of the information and how it may affect relationships between patients and doctors. Little empirical research has examined how the internet is used by those diagnosed with a serious illness. This study uses in-depth interviews collected for DIPEx projects with men and women with cancer. The analysis is informed by Radley and Billig's (Sociol. Health Illness 2 (1996) 220) observation that accounts of illness require patients to simultaneously display themselves as 'worthy individuals, as more or less fit participants in the social world' and on Arthur Frank's work on quest narratives. Drawing on in-depth interviews with a woman with inflammatory breast cancer and a man with prostate cancer I will demonstrate how the internet has been used not only to gather information and gain support from others but also to make sense of the experience of cancer. The ability to access health information on the internet may provide patients with an opportunity to display a particularly modern marker of competence and social fitness. However, one of the consequences of easier access to health information may be the emergence of a felt imperative to be (or present oneself as) an expert and critical patient, able to question advice and locate effective treatments for oneself.


Real-space observation of molecular motion induced by femtosecond laser pulses.
Femtosecond laser irradiation is used to excite adsorbed CO molecules on a Cu110 surface; the ensuing motion of individual molecules across the surface is characterized on a site-to-site basis by in situ scanning tunneling microscopy. Adsorbate motion both along and perpendicular to the rows of the Cu110 surface occurs readily, in marked contrast to the behavior seen for equilibrium diffusion processes. The experimental findings for the probability and direction of the molecular motion can be understood as a manifestation of strong coupling between the adsorbates' lateral degrees of freedom and the substrate electronic excitation produced by the femtosecond laser radiation.


Aiding diagnosis of childhood attention-deficit/hyperactivity disorder of the inattentive presentation: Discriminant function analysis of multi-domain measures including EEG.
We developed a neurocognitive assessment tool (NCAT) in consultation with mental health professionals working with children with AD/HD as a diagnostic aid and screening tool. This study examines the predictive utility of NCAT in the classification of children with AD/HD Inattentive presentation. Fifty three children with AD/HD Inattentive presentation and 161 typically-developing children completed an NCAT assessment. Discriminant function analyses examined group membership prediction for separate components of NCAT and for the components combined. The combined model correctly classified 93.4 % of participants, with 91.4 % sensitivity and 93.9 % specificity. Contributions to classification were from SNAP-IV, psychological needs satisfaction, self-regulation, executive function performance, and EEG. The combined model resulted in a 9.3 % increase in specificity and 5.9 % increase in sensitivity compared to SNAP-IV alone. NCAT provides good discrimination between children with and without AD/HD of the Inattentive presentation, and further investigation including other subtypes and comorbidities is warranted.


Amisulpride Prevents Postoperative Nausea and Vomiting in Patients at High Risk: A Randomized, Double-blind, Placebo-controlled Trial.
Postoperative nausea and vomiting causes distress for patients and can prolong care requirements. Consensus guidelines recommend use of multiple antiemetics from different mechanistic classes as prophylaxis in patients at high risk of postoperative nausea and vomiting. The prophylactic efficacy of the dopamine D2/D3 antagonist amisulpride in combination with other antiemetics was investigated. This double-blind, randomized, placebo-controlled, international, multicenter trial was conducted in 1,147 adult surgical patients having three or four postoperative nausea and vomiting risk factors. Patients were randomized to receive either intravenous amisulpride (5 mg) or matching placebo at induction of general anesthesia, in addition to one standard, nondopaminergic antiemetic, most commonly ondansetron or dexamethasone. Vomiting/retching, nausea, and use of rescue medication were recorded for 24 h after wound closure. The primary endpoint was complete response, defined as no emesis or rescue medication use in the 24-h postoperative period. Complete response occurred in 330 of 572 (57.7%) of the amisulpride group and 268 of 575 (46.6%) of the control group (difference 11.1 percentage points; 95% CI, 5.3 to 16.8; P < 0.001). The incidences of emesis (13.8% vs. 20.0%, P = 0.003), any nausea (50.0% vs. 58.3%, P = 0.002), significant nausea (37.1% vs. 47.7%, P < 0.001), and rescue medication use (40.9% vs. 49.4%, P = 0.002) were significantly lower in the amisulpride group. Adverse events and laboratory and electrocardiogram abnormalities occurred no more frequently with amisulpride than with placebo. Intravenous amisulpride was safe and effective as prophylaxis of postoperative nausea and vomiting when given in combination with an antiemetic from another class to adult patients at high risk for suffering postoperative nausea and vomiting undergoing elective surgery under inhalational general anesthesia. An online visual overview is available for this article at http://links.lww.com/ALN/B727.


Immune functions of proteinase 3.
The primary function of neutrophil-derived serine proteases, neutrophil elastase, cathepsin G, and proteinase 3 (PR3) are thought to be the degradation of extracellular proteins at sites of inflammation, but excessive, prolonged, or inappropriate proteolytic activity causes harmful effects in the body. Although there are strong structural similarities among these proteases, PR3 has unique properties in many respects. In particular, PR3 is a major target antigen of autoantibodies, anti-neutrophil cytoplasmic antibodies (ANCA). Recent findings also revealed that PR3 is critically involved in the regulation of immune function. This review discusses the expression of PR3 in non-hemopoietic cells and focuses on the immune functions of PR3 with respect to direct modulation of cell signaling by PR3, PR3-mediated cell activation, and the possible involvement of protease-activated receptors, modulation of the cytokine network, innate immunity and PR3, and recent findings about the generation and function of ANCA.


Isolation by environment in the highly mobile olive ridley turtle (Lepidochelys olivacea) in the eastern Pacific.
Spatial and temporal scales at which processes modulate genetic diversity over the landscape are usually overlooked, impacting the design of conservation management practices for widely distributed species. We examine processes shaping population divergence in highly mobile species by re-assessing the case of panmixia in the iconic olive ridley turtle from the eastern Pacific. We implemented a biophysical model of connectivity and a seascape genetic analysis based on nuclear DNA variation of 634 samples collected from 27 nesting areas. Two genetically distinct populations largely isolated during reproductive migrations and mating were detected, each composed of multiple nesting sites linked by high connectivity. This pattern was strongly associated with a steep environmental gradient and also influenced by ocean currents. These findings relate to meso-scale features of a dynamic oceanographic interface in the eastern tropical Pacific (ETP) region, a scenario that possibly provides different cost-benefit solutions and selective pressures for sea turtles during both the mating and migration periods. We reject panmixia and propose a new paradigm for olive ridley turtles where reproductive isolation due to assortative mating is linked to its environment. Our study demonstrates the relevance of integrative approaches for assessing the role of environmental gradients and oceanographic currents as drivers of genetic differentiation in widely distributed marine species. This is relevant for the conservation management of species of highly mobile behaviour, and assists the planning and development of large-scale conservation strategies for the threatened olive ridley turtles in the ETP.


Coalesced poly(ε-caprolactone) fibers are stronger.
Melt-spun fibers were made from poly(ε-caprolactone) (PCL) coalesced from stoichiometric inclusion complex crystals formed with host urea. Melting and crystallization behaviors, mechanical properties, and the birefringence of undrawn and cold-drawn fibers were investigated. Undrawn coalesced PCL fibers were observed to have 500-600% higher moduli than undrawn as-received (asr) PCL fibers and a modulus comparable to drawn asr PCL fibers. Drawn coalesced PCL fibers have the highest crystallinity, orientation, and 65% higher moduli than drawn asr PCL fibers. Drawn coalesced PCL fibers have only a 5% higher crystallinity than drawn asr PCL fibers, yet they have 65% higher moduli and lower elongation at break values. Clearly, the intrinsic alignment of the coalesced polymers is the reason for their higher moduli and lower elongation, as confirmed by the birefringence observed in drawn coalesced and asr-PCL fibers. The improved mechanical properties of coalesced PCL fibers make them a better candidate for use in tissue engineering as scaffolds.


Obvious temperature difference along a pb cluster-decorated carbon nanowire.
Pb nanoclusters were deposited onto a suspended carbon nanowire (CNW), where in situ temperature variable observation was carried out by a transmission electron microscope. The heating temperature was up to 450 °C. Both the melting and evaporation of the Pb nanoparticles on the CNW were retarded when compared to the particles on the support frame. The obvious temperature difference of up to 10 K along the CNW of less than 1 μm was demonstrated. It was attributed to the irradiating dissipation-dependent on the surface area of the decorating Pb particle by calculation.(See supplementary material 1).


The upper limb of Homo naledi.
The evolutionary transition from an ape-like to human-like upper extremity occurred in the context of a behavioral shift from an upper limb predominantly involved in locomotion to one adapted for manipulation. Selection for overarm throwing and endurance running is thought to have further shaped modern human shoulder girdle morphology and its position about the thorax. Homo naledi (Dinaledi Chamber, Rising Star Cave, Cradle of Humankind, South Africa) combines an australopith-like cranial capacity with dental characteristics akin to early Homo. Although the hand, foot, and lower limb display many derived morphologies, the upper limb retains many primitive traits. Here, we describe the H. naledi upper extremity (excluding the hand) in detail and in a comparative context to evaluate the diversity of clavicular, scapular, humeral, radial, and ulnar morphology among early hominins and later Homo. Homo naledi had a scapula with a markedly cranially-oriented glenoid, a humerus with extremely low torsion, and an australopith-like clavicle. These traits indicate that the H. naledi scapula was situated superiorly and laterally on the thorax. This shoulder girdle configuration is more similar to that of Australopithecus and distinct from that of modern humans, whose scapulae are positioned low and dorsally about the thorax. Although early Homo erectus maintains many primitive clavicular and humeral features, its derived scapular morphology suggests a loss of climbing adaptations. In contrast, the H. naledi upper limb is markedly primitive, retaining morphology conducive to climbing while lacking many of the derived features related to effective throwing or running purported to characterize other members of early Homo.


A preliminary study of the effects of repeated massage on hypothalamic-pituitary-adrenal and immune function in healthy individuals: a study of mechanisms of action and dosage.
This study gathers preliminary data about the biologic effects of repeated Swedish massage therapy compared to a light-touch control condition. The study design was a 5-week comparison of repeated Swedish massage and light touch on oxytocin (OT), arginine-vasopressin (AVP), adrenal corticotropin hormone (ACTH), cortisol (CORT), circulating phenotypic lymphocyte markers, and mitogen-stimulated cytokine function. The setting was an outpatient research unit in an academic medical center. The study subjects were medically and psychiatrically healthy young adults. The study comprised 45 minutes of Swedish massage or light touch, using highly specified and identical protocols, either weekly or twice weekly for 5 weeks. The outcome measures were mean differences between massage and light touch on OT, AVP, ACTH, CORT, lymphocyte markers, and cytokine levels. Compared to the touch control condition, weekly Swedish massage stimulated a sustained pattern of increased circulating phenotypic lymphocyte markers and decreased mitogen-stimulated cytokine production, similar to what was previously reported for a single massage session, while having minimal effect on hypothalamic-pituitary-adrenal function. Twice-weekly massage produced a different response pattern with increased OT levels, decreased AVP, and decreased CORT but little effect on circulating lymphocyte phenotypic markers and a slight increase in mitogen-stimulated interferon-γ, tumor necrosis factor-α, interleukin (IL)-1b and IL-2 levels, suggesting increased production of pro-inflammatory cytokines. There are sustained cumulative biologic actions for the massage and touch interventions that persist for several days or a week, and these differ profoundly depending on the dosage (frequency) of sessions. Confirmatory studies in larger samples are needed.


The role of exercise in the management of adverse effects of androgen deprivation therapy for prostate cancer: a rapid review.
Prostate cancer (PCa) is the most commonly diagnosed cancer in Australia, accounting for one quarter of all new cancer diagnoses for males. Androgen deprivation therapy (ADT) is the standard first-line therapy for metastatic PCa but is also used across much of the spectrum of disease. Unfortunately, debilitating adverse effects are a significant and largely unavoidable feature of ADT. A recent systematic review of adverse effects of ADT identified 19 sub-groups classified according to the Common Terminology Criteria for Adverse Events (CTCAE) Version 5.0. The potential for multiple simultaneous adverse effects, their associated management and the impact of adverse effects on cancer outcomes and quality of life are important considerations in the treatment and supportive care of men with PCa. Exercise is increasingly being recognized as an efficacious strategy in managing these adverse effects. A rapid review was undertaken to examine the role of exercise in the management of the most commonly reported ADT adverse effects classified according to the CTCAE sub-groups. A systematic search was conducted in Medline, PsycINFO, Google Scholar and Google for the years 2010 to September 2019 to identify the benefits of exercise in managing the adverse effects of ADT for PCa. There is strong evidence for exercise as medicine in addressing several of the adverse effects of PCa such as loss of muscle mass and strength, fatigue and declining physical function. Moderate level evidence for PCa exists for exercise-induced improvements in depression and anxiety, bone loss, and sexual dysfunction. While evidence of the effectiveness of exercise is lacking for many adverse effects of ADT for PCa, evidence in the cancer population as a whole or other clinical populations is strong, and many clinical guidelines recommend exercise as a fundamental part of their clinical management. With the exception of gynaecomastia and breast pain, there is increasing evidence (PCa, cancer or other clinical populations) to suggest that exercise has the potential to reduce and even prevent many of the adverse effects of ADT, thus improving survivorship outcomes for men with PCa. Exercise has the potential to reduce and even prevent many of the adverse effects of ADT, thus improving survivorship outcomes for men with PCa. The use of exercise for PCa management has the potential to translate into health and economic benefits in improved quality of life and fewer complications, resulting in savings to the health care system, enhanced productivity and reduced patient and carer burden. Exercise thus has the potential to improve quality of life for this population as well as generate significant cost savings.


Factors associated with intention to engage in self-protective behavior: The case of over-the-counter acetaminophen products.
Inappropriate use of acetaminophen products is a concern due to the severe liver damage associated with intentional or accidental overdose of these products. In 2009, the U.S. Food and Drug Administration (FDA) issued more severe organ-specific warnings for the acetaminophen Drug Facts label to improve protective behavior among patients. However, it is not clear how patients react to such interventions by the FDA. The objective of this study was to evaluate the factors influencing patients' intention to engage in protective behavior while using acetaminophen products after reading the Drug Facts label. The study specifically looked at the relationship between four Protection Motivation Theory-based risk cognition factors and the intention to engage in protective behavior. An experimental, cross-sectional, field study was conducted using self-administered questionnaires at four community pharmacies in Houston, TX. Two hundred surveys were collected from adults visiting the selected pharmacy stores. Participants were exposed to a simulated label (i.e. Drug Facts label) containing organ-specific warnings for over-the-counter (OTC) acetaminophen products. Risk cognition measures (i.e. measures of perceived severity, perceived vulnerability, response efficacy, and self-efficacy) and measures of intention to engage in protective behavior (always reading warnings, using products with more caution, and consulting a pharmacist/physician) were recorded. Pearson correlation and multiple linear regression analyses, controlling for demographic and behavioral characteristics of the participants, were performed. Bivariate analyses indicated that an increase in perceived severity, perceived vulnerability and response efficacy were associated with a higher intention to engage in protective behavior. Findings from the multiple regression indicated that increase in perceived severity of liver damage, belonging to a non-healthcare occupation, no history of acetaminophen use and no history of alcohol consumption were associated with a higher intention to engage in protective behavior. Higher risk cognition of liver damage associated with inappropriate use of OTC acetaminophen products leads to greater intention to engage in protective behavior while using such products. Developing interventions targeted towards improving reading and adhering to the Drug Facts label could improve risk cognition, and thus improve patients' intention to engage in protective behavior. Regular acetaminophen users, heavy alcohol consumers and healthcare professionals might need other interventions apart from the Drug Facts label to improve their likelihood to engage in protective behavior.


Practical oxazole synthesis mediated by iodine from α-bromoketones and benzylamine derivatives.
The reagent system of I2/K2CO3 could efficiently promote the oxazole synthesis from α-bromoketones and benzylamine derivatives in DMF. This method was not only suitable for 2,5-diaryl oxazole synthesis but also for 2,4,5-trisubstituted oxazole and 5-alkyl/alkenyl oxazole synthesis. Furthermore, this method was successfully applied to a one-step synthesis of a natural product halfordinol in 62% yield.


Conformational changes in the Ebola virus membrane fusion machine induced by pH, Ca2+, and receptor binding.
The Ebola virus (EBOV) envelope glycoprotein (GP) is a membrane fusion machine required for virus entry into cells. Following endocytosis of EBOV, the GP1 domain is cleaved by cellular cathepsins in acidic endosomes, removing the glycan cap and exposing a binding site for the Niemann-Pick C1 (NPC1) receptor. NPC1 binding to cleaved GP1 is required for entry. How this interaction translates to GP2 domain-mediated fusion of viral and endosomal membranes is not known. Here, using a bulk fluorescence dequenching assay and single-molecule Förster resonance energy transfer (smFRET)-imaging, we found that acidic pH, Ca2+, and NPC1 binding synergistically induce conformational changes in GP2 and permit virus-liposome lipid mixing. Acidic pH and Ca2+ shifted the GP2 conformational equilibrium in favor of an intermediate state primed for NPC1 binding. Glycan cap cleavage on GP1 enabled GP2 to transition from a reversible intermediate to an irreversible conformation, suggestive of the postfusion 6-helix bundle; NPC1 binding further promoted transition to the irreversible conformation. Thus, the glycan cap of GP1 may allosterically protect against inactivation of EBOV by premature triggering of GP2.


Extracellular secretion of Escherichia coli heat-stable enterotoxin I across the outer membrane.
Escherichia coli heat-stable enterotoxin Ip (STIp) is an extracellular toxin consisting of 18 amino acid residues that is synthesized as a precursor of pre (amino acid residues 1 to 19), pro (amino acid residues 20 to 54), and mature (amino acid residues 55 to 72) regions. The precursor synthesized in the cytoplasm is translocated across the inner membrane by the general export pathway consisting of Sec proteins. The pre region functions as a leader peptide and is cleaved during translocation. However, it remains unknown how the resulting peptide (pro-mature peptide) translocates across the outer membrane. In this study, we investigated the structure of the STIp that passes through the outer membrane to determine how it translocates through the outer membrane. The results showed that the pro region is cleaved in the periplasmic space. The generated peptide becomes the mature form of STIp, which happens to have disulfide bonds, which then passes through the outer membrane. We also showed that STIp with a carboxy-terminal peptide consisting of 3 amino acid residues passes through the outer membrane, whereas STIp with a peptide composed of 37 residues does not. Amino acid analysis of mutant STIp purified from culture supernatant revealed that the peptide composed of 37 amino acid residues was cleaved into fragments of 5 amino acid residues. In addition, analyses of STIps with a mutation at the cysteine residue and the dsbA mutant strain revealed that the formation of an intramolecular disulfide bond within STIp is not absolutely required for the mature region of STIp to pass through the outer membrane.


Effects of citral, a naturally occurring antiadipogenic molecule, on an energy-intense diet model of obesity.
Retinaldehyde inhibits adipogenesis, increases metabolic rate, reduces weight gain, and improves tolerance to a glucose load. We assessed the effects of citral - an inhibitor of retinaldehyde dehydrogenase (the primary enzyme metabolizing retinaldehyde), on body weight, glucose tolerance, fasting plasma glucose and insulin levels, metabolic rate, adipocyte size, and morphology in a diet-induced model of obesity. Out of the 5 groups of 6-week-old male Sprague-Dawley rats, 4 were maintained on an energy-intense, palatable, diet for a period of - 42 days, while 1 served as the control. After obesity had been induced, 3 groups were treated with daily doses of citral (10, 15, and 20 mg/kg body weight) for a period of 28 days. They were then subjected to metabolic experiments. Body weight, fasting plasma glucose, glucose tolerance to an intraperitoneal glucose load, metabolic rate, and adipocyte size were assessed. Citral-treated groups showed a dose-dependent reduction in body weight gain. They significantly had lower fasting glucose levels, improved glucose tolerance, lower fasting plasma glucose, higher metabolic rate, and smaller adipocytes after drug administration. The findings suggest that citral increased energy dissipation (and also reduced lipid accumulation) consequently preventing and ameliorating diet-induced obesity. In addition it improved insulin sensitivity and glucose tolerance. In the current scenario of increasing prevalence of obesity and diabetes, citral may prove as novel agent in its management.


Antioxidant, antibacterial and ACE-inhibitory activity of four monofloral honeys in relation to their chemical composition.
Different monofloral honeys from Castilla-La Mancha (Spain) have been studied in order to determine their main functional and biological properties. Thyme honey and chestnut honey possess the highest antioxidant capacity, which is due to their high vitamin C (in thyme honey) and total polyphenolic content (in chestnut honey). On the other hand, chestnut honey showed high antimicrobial activity against Staphylococcus aureus and Escherichia coli, whilst others had no activity against S. aureus and showed very small activity against E. coli. Moreover it was found that the antimicrobial activity measured in chestnut honey was partly due to its lysozyme content. In addition the angiotensin I-converting enzyme (ACE) inhibitory activity was measured, and the ACE inhibition is one mechanism by which antihypertensive activity is exerted in vivo. All the types of honey showed some activity but chestnut honey had the highest ACE inhibitory activity.


Using insurance standards and policy levers to build a high performance health system.
This issue brief examines an unprecedented use of state health insurance regulatory authority to promote health system reform. In 2004, the Rhode Island legislature created the Office of the Health Insurance Commissioner (OHIC) with authority not granted to state health insurance regulatory agencies in other states. Specifically, the legislation instructed OHIC to direct insurers toward policies that promote improved accessibility, quality, and affordability for the Rhode Island health system. In 2009, OHIC used this authority to implement a set of standards to promote increased affordability through a series of requirements aimed at strengthening and expanding the state's primary care infrastructure. Insurers are required to increase their investments in primary care on a cost-neutral basis, expand use of the chronic care model medical home, and support implementation of electronic medical records. Rhode Island is testing whether state insurance regulation can foster a profound transformation in health care delivery.


Two types of thalamocortical projections from the motor thalamic nuclei of the rat: a single neuron-tracing study using viral vectors.
The axonal arborization of single motor thalamic neurons was examined in rat brain using a viral vector expressing membrane-targeted palmitoylation site-attached green fluorescent protein (palGFP). We first divided the ventral anterior-ventral lateral motor thalamic nuclei into 1) the rostromedial portion, which was designated inhibitory afferent-dominant zone (IZ) with intense glutamate decarboxylase immunoreactivity and weak vesicular glutamate transporter 2 immunoreactivity, and 2) the caudolateral portion, named excitatory subcortical afferent-dominant zone (EZ) with the reversed immunoreactivity profile. We then labeled 38 motor thalamic neurons in 29 hemispheres by injecting a diluted palGFP-Sindbis virus solution and isolated 10 IZ and EZ neurons for reconstruction. All the reconstructed IZ neurons widely projected not only to the cerebral cortex but also to the neostriatum, whereas the EZ neurons sent axons almost exclusively to the cortex. More interestingly, 47-66% of axon varicosities of IZ neurons were observed in layer I of cortical areas. In contrast, only 2-15% of varicosities of EZ neurons were found in layer I, most varicosities being located in middle layers. These results suggest that 2 forms of information from the basal ganglia and cerebellum are differentially supplied to apical and basal dendrites, respectively, of cortical pyramidal neurons and integrated to produce a motor execution command.


Echocardiographic evidence of left atrial mechanical dysfunction after conversion of atrial fibrillation to sinus rhythm in 5 horses.
Atrial contractile dysfunction occurs in some species after conversion of atrial fibrillation (AF) to normal sinus rhythm (NSR) but has not been reported in horses with naturally occurring AF. Transthoracic echocardiography allows detection of left atrial (LA) mechanical dysfunction in horses after conversion of AF to NSR. Five Standardbreds with AF and 6 healthy Standardbreds of similar age, weight, and athletic condition were included in this study. Four horses were treated pharmacologically (quinidine), and 1 horse was treated by means of transvenous electrical cardioversion. Echocardiographic examinations were performed in normal horses (once) and in AF horses (24 hours and 72 hours after conversion to NSR) by means of 2-dimensional echocardiography (2DE), transmitral flow Doppler, and tissue Doppler imaging (TDI) techniques. Echocardiographic indices of LA mechanical function were compared between normal horses and AF horses. Two-dimensional echocardiography and TDI indices of LA mechanical function revealed significant decreases in LA contractile function and LA reservoir function 24 hours after cardioversion. This decrease was no longer statistically significant 72 hours after cardioversion, but changes in echocardiographic variables between 24 and 72 hours varied among horses. LA contractile dysfunction can be evaluated in horses by use of 2DE, transmitral Doppler flow velocity profiles, and analyses of LA wall motion by TDI. The results of this study are consistent with AF-induced atrial remodeling, although residual treatment effects or influence of underlying primary myopathy cannot be excluded.


Mental health nurses' experiences of caring for patients suffering from self-harm.
The aim of this study was to explore mental health nurses' experiences of caring for inpatients who self-harm during an acute phase. The setting was four psychiatric clinics in Norway. Fifteen mental health nurses (MHNs) were recruited. Semistructured interviews comprised the method for data collection, with content analysis used for data analysis. Two main categories emerged: challenging and collaborative nurse-patient relationship and promoting well-being through nursing interventions. The underlying meaning of the main categories was interpreted and formulated as a latent theme: promoting person-centered care to patients suffering from self-harm. How MHNs promote care for self-harm patients can be described as a person-centered nursing process. MHNs, through the creation of a collaborative nurse-patient relationship, reflect upon nursing interventions and seek to understand each unique patient. The implication for clinical practice is that MHNs are in a position where they can promote patients' recovery processes, by offering patients alternative activities and by working in partnership with patients to promote their individual strengths and life knowledge. MHNs strive to help patients find new ways of living with their problems. The actual study highlighted that MHNs use different methods and strategies when promoting the well-being of self-harm patients.


Target protectors reveal dampening and balancing of Nodal agonist and antagonist by miR-430.
MicroRNAs (miRNAs) repress hundreds of target messenger RNAs (mRNAs), but the physiological roles of specific miRNA-mRNA interactions remain largely elusive. We report that zebrafish microRNA-430 (miR-430) dampens and balances the expression of the transforming growth factor-beta (TGF-beta) Nodal agonist squint and the TGF-beta Nodal antagonist lefty. To disrupt the interaction of specific miRNA-mRNA pairs, we developed target protector morpholinos complementary to miRNA binding sites in target mRNAs. Protection of squint or lefty mRNAs from miR-430 resulted in enhanced or reduced Nodal signaling, respectively. Simultaneous protection of squint and lefty or absence of miR-430 caused an imbalance and reduction in Nodal signaling. These findings establish an approach to analyze the in vivo roles of specific miRNA-mRNA pairs and reveal a requirement for miRNAs in dampening and balancing agonist/antagonist pairs.


Understanding the contribution of habitats and regional variation to long-term population trends in tricolored blackbirds.
Population trends represent a minimum amount of information required to assess the conservation status of a species. However, understanding and detecting trends can be complicated by variation among habitats and regions, and by dispersal connecting habitats through source-sink dynamics. We analyzed trends in breeding populations between habitats and regions to better understand the overall dynamics of a species' decline. Specifically, we analyzed historical trends in breeding populations of tricolored blackbirds (Agelaius tricolor) using breeding records from 1907 to 2009. The species breeds itinerantly and ephemerally uses multiple habitat types and breeding areas, which make interpretation of trends complex. We found overall abundance declines of 63% between 1935 and 1975. Since 1980 overall declines became nonsignificant and obscure despite large amounts of data from 1980 to 2009. Temporal trends differed between breeding habitat types and were associated with regional differences in population declines. A new habitat, triticale crops (a wheat-rye hybrid grain) produced colonies 40× larger, on average, than other breeding habitats, and contributed to a change in regional distribution since it primarily occurred in a single region. The mechanism for such an effect is not clear, but could represent the local availability of foodstuffs in the landscape rather than something specific to triticale crops. While variation in trends among habitats clearly occurred, they could not easily be ascribed to source-sink dynamics, ecological traps, habitat selection or other detailed ecological mechanisms. Nonetheless, such exchanges provide valuable information to guide management of dynamic systems.


Different pathophysiology underlying animal models of fibromyalgia and neuropathic pain: comparison of reserpine-induced myalgia and chronic constriction injury rats.
The reserpine-induced myalgia (RIM) rat manifests fibromyalgia-like chronic pain symptoms. The present study explored the pathophysiology underlying the pain symptoms in the RIM rat and the chronic constriction injury (CCI) rat, an animal model of neuropathic pain as a reference. Nerve tissue samples were collected from the nociception-tested animals for pathological examinations. Additionally, the therapeutic efficacy of a sodium channel blocker mexiletine was assessed in both rats. A slight vacuolization in the substantia nigra (SN) occurred in some of the RIM rats without any other histopathological changes in the brain or peripheral neurons. All the RIM rats, with or without vacuolization, showed hypersensitivity to tactile, muscle pressure, and cold stimuli. In the CCI rat, neurodegenerative changes were apparent in the sciatic nerve and the spinal cord only. CCI rats displayed muscle hyperalgesia in addition to tactile and cold allodynia. Pharmacotherapy with mexiletine did not attenuate the pain in the RIM rat, although it was effective in the CCI rat. Taken together, it is not likely that pain symptoms in RIM rats are caused by degenerative changes at the level of primary afferents and spinal cord, as is the case for CCI rats. The significance of the vacuolization in the SN is less clear at present because of the minor extent of the change and the lack of correlation with nociceptive sensitivity. The pain symptoms in RIM rats could be associated with dysfunction of biogenic amines-mediated CNS pain control even without apparent pathologies in the nervous system.


A new hero emerges: another exceptional mammalian spine and its potential adaptive significance.
The hero shrew's (Scutisorex somereni) massive interlocking lumbar vertebrae represent the most extreme modification of the vertebral column known in mammals. No intermediate form of this remarkable morphology is known, nor is there any convincing theory to explain its functional significance. We document a new species in the heretofore monotypic genus Scutisorex; the new species possesses cranial and vertebral features representing intermediate character states between S. somereni and other shrews. Phylogenetic analyses of DNA sequences support a sister relationship between the new species and S. somereni. While the function of the unusual spine in Scutisorex is unknown, it gives these small animals incredible vertebral strength. Based on field observations, we hypothesize that the unique vertebral column is an adaptation allowing these shrews to lever heavy or compressive objects to access concentrated food resources inaccessible to other animals.


Inconsistent reporting of female genital cutting status in northern Ghana: explanatory factors and analytical consequences.
Although many cross-sectional social surveys have included questions about female genital cutting status and correlated personal characteristics, no longitudinal studies have been launched that permit investigation of response biases associated with such surveys. This study draws upon the findings of a longitudinal study of women aged 15 to 49 in rural northern Ghana. The self-reported circumcision status of women interviewed in 1995 was compared with the status they reported when they were interviewed again in 2000 after the government began enforcing a law banning the practice and public information campaigns against it were launched. In all, 13 percent of respondents who reported in 1995 that they had been circumcised stated that they had not been circumcised in the 2000 reinterview; this inconsistency reached 50 percent for the youngest age group. Analysis shows that women who said they had not been circumcised are significantly younger, more likely to be educated, and less likely to practice traditional religion than are women who reported that they were circumcised. Factors that may explain these correlates of denial are discussed, and implications for research are reviewed.


The epidemiology of oro-nasal haemorrhage and suffocation in infants admitted to hospital in Scotland over 10 years.
To estimate the incidence of oro-nasal haemorrhage (ONH) and suffocation in infancy and to investigate their aetiology and overlap. Setting A 10-year retrospective hospital based study from Scotland, UK. The hospital notes of all infants presenting with ONH or suffocation identified through the Information Services Division of the Scottish Health Service were reviewed by three paediatric consultants, two with child protection expertise. The hospital-based incidences of haemorrhages from different sites were calculated and the causes ascertained. When trauma was involved, a decision was made whether it was likely to have been accidental. 7 cases of suffocation and 88 of ONH were recorded at hospital discharge over 10 years. This gives an incidence of ONH of 1.62 (1.30 to 1.99)/10,000 live births (95% CI) which consists of haemorrhage arising from nose or mouth (N/M), n=65 (1.19/10,000 (0.92 to 1.52)); haematemesis, n=11 (0.20/10,000 (0.10 to 0.36)); haemoptysis, n=3 (0.06/10,000 (0.1 to 0.16)) and pulmonary haemorrhage, n=9 (0.17/10,000 (0.08 to 0.31)). No suffocation cases were recorded as having a coincident ONH, but five ONH cases were probably caused by airway obstruction. 40 of 65 cases of N/M were associated with trauma, which in 15 cases were thought to be probable abuse; four were associated with coagulation abnormalities. 2/3 haemoptysis cases, 2/11 haematemesis cases and 8/65 N/M cases were associated with a coincident respiratory tract infection, though in 4/8 of these cases, there was an associated apparent life-threatening event. Haemorrhage from the N/M is rare in infancy. Trauma is commonly involved and child protection concerns often poorly explored. Pulmonary haemorrhage and several cases of ONH were associated with probable airway obstruction. Information, in cases of ONH, is in general recorded badly, and an investigation and management plan are suggested.


Epigenetically reprogramming of human embryonic stem cells by 3-deazaneplanocin a and sodium butyrate.
Infertility affects about 6.1 million women aged 15-44 in the United States. The leading cause of infertility in women is quantitative and qualitative defects in human germ-cell development (these sentences are not mentioned in introduction so it is not correct to mention in abstract, you can omit). Human embryonic stem cell (hESC) lines are derived from the inner cell mass (ICM) of developing blastocysts and have a broad clinical potential. hESCs have been classified into three classes based on their epigenetic state. The goal of this study was to epigenetically reprogram Class II and Class III cell lines to Class I (naïve state), and to in vitro differentiation of potent hESCs to primordial germ cells (PGCs). Recent evidence suggests that 3-deazaneplanocin A (DZNep) is a global histone methylation inhibitor which selectively inhibits trimethylation of lysine 27 on histone H3K27, and it is an epigenetic therapeutic for cancer. The characteristics of DZNep lead us to hypothesize that it is a good candidate to epigenetically reprogram hESCs to the Class I. Additionally, we used sodium butyrate (NaBu) shown in previous studies to up-regulate the expression of germ cell specific markers (these sentences should be come in introduction). We used these two drugs to produce epigenetically stable hESC lines. hESC lines are an appropriate system for disease modeling and understanding developmental stages, therefore producing stable stem cell lines may have an outstanding impact in different research fields such as preventive medicine. X-Chromosome inactivation has been used as a tool to follow the reprogramming process. We have used immunostaining and western blot as methods to follow this reprogramming qualitatively and quantitatively.


The relative efficiency of beta adrenoceptor coupling to myocardial inotropy and diastolic relaxation: organ-selective treatment for diastolic dysfunction.
The relative effects of drugs which elevate cytosolic cyclic AMP on inotropy and diastolic relaxation (lusitropy) of guinea pig atria were quantified in vitro. There was a temporal difference between these responses in that inotropy reached peak response considerably faster than lusitropy. Also, although the relaxation response was sustained to an elevated steady state, the inotropic responses to beta adrenoceptor agonists were transient and returned to base line over 90 min. However, the inotropic responses to forskolin and dibutyryl cyclic AMP (cAMP) were sustained. For all of the drugs tested, the lusitropic response was at least 4 times more sensitive than the inotropic response (i.e., the concentration response curve for relaxation was shifted to the left of the curve for inotropy). In the case of beta adrenoceptor agonists, these differences were greater, presumably because of the fading inotropic response over 90 min. It was found that although high efficacy beta adrenoceptor agonists such as isoproterenol (and the direct activator of adenylate cyclase forskolin) produced both inotropy and lusitropy, lower efficacy agonists produced predominant lusitropy. The low efficacy agonist prenalterol produced insignificant inotropy but 60% maximal lusitropy. These data were modeled mathematically by a "differential coupling model" which assumed that a uniform cytosolic level of elevated cAMP activated two biochemical processes of differing sensitivity. Thus, the lusitropic response (phosphorylation of phospholamban) was coupled more efficiently to the cAMP response than the inotropic response (phosphorylation of calcium channels). A second model ("differential messenger concentration model") which calculated the effects of a compartmentalization of cAMP concentration within the cardiac cell by restricted diffusion and/or selective degradation by phosphodiesterases also was used.(ABSTRACT TRUNCATED AT 250 WORDS)


An analysis of policies for cotrimoxazole, amoxicillin and azithromycin use in Namibia's public sector: Findings and therapeutic implications.
Despite Namibia's robust medicine use systems and policies, antibiotic use indicators remain suboptimal. Recent medicine use surveys rank cotrimoxazole, amoxicillin and azithromycin (CAA) among the most used medicines. However, there is rising resistance to CAA (55.9%-96.7%). Unfortunately, to date, there have been limited studies evaluating policies to improve antibiotic use in Namibia. To evaluate public sector pharmaceutical policies and guidelines influencing the therapeutic use of CAA antibiotics in Namibia. Evaluate Namibia's pharmaceutical policies and guidelines for CAA use through quantitative text analysis. The main outcome variables were the existence of antibiotic policies, therapeutic indications per antibiotic and the type/level of healthcare facility allowed to use the antibiotic. Policies for antibiotic use were limited, with only the draft Namibia Medicines Policy having a statement on antibiotic use. Several essential antibiotics had no therapeutic indications mentioned in the guidelines. Twenty-nine antibiotics were listed for 69 therapeutic indications; CAA (49.3%) antibiotics and ATC J01C/J01D (48%) having the highest indications per antibiotic. For CAA antibiotics, this suggested use was mainly for acute respiratory infections (n=22, 37.2%). Published policies (58.6%-17/29) recommended antibiotics for use at the primary healthcare (PHC) level, with CAA antibiotics recommended mostly for respiratory tract infections and genitourinary infections. Policy and guidelines for antibiotic use in Namibia are not comprehensive and are skewed towards PHCs. Existing policies promote the wide use of CAA antibiotics, which may inadvertently result in their inappropriate use enhancing resistance rates. This calls for the development of more comprehensive antibiotic guidelines and essential medicine lists in tandem with local antimicrobial resistance patterns. In addition, educational initiatives among all key stakeholder groups.


Root morphology and seed and leaf ionomic traits in a Brassica napus L. diversity panel show wide phenotypic variation and are characteristic of crop habit.
Mineral nutrient uptake and utilisation by plants are controlled by many traits relating to root morphology, ion transport, sequestration and translocation. The aims of this study were to determine the phenotypic diversity in root morphology and leaf and seed mineral composition of a polyploid crop species, Brassica napus L., and how these traits relate to crop habit. Traits were quantified in a diversity panel of up to 387 genotypes: 163 winter, 127 spring, and seven semiwinter oilseed rape (OSR) habits, 35 swede, 15 winter fodder, and 40 exotic/unspecified habits. Root traits of 14 d old seedlings were measured in a 'pouch and wick' system (n = ~24 replicates per genotype). The mineral composition of 3-6 rosette-stage leaves, and mature seeds, was determined on compost-grown plants from a designed experiment (n = 5) by inductively coupled plasma-mass spectrometry (ICP-MS). Seed size explained a large proportion of the variation in root length. Winter OSR and fodder habits had longer primary and lateral roots than spring OSR habits, with generally lower mineral concentrations. A comparison of the ratios of elements in leaf and seed parts revealed differences in translocation processes between crop habits, including those likely to be associated with crop-selection for OSR seeds with lower sulphur-containing glucosinolates. Combining root, leaf and seed traits in a discriminant analysis provided the most accurate characterisation of crop habit, illustrating the interdependence of plant tissues. High-throughput morphological and composition phenotyping reveals complex interrelationships between mineral acquisition and accumulation linked to genetic control within and between crop types (habits) in B. napus. Despite its recent genetic ancestry (<10 ky), root morphology, and leaf and seed composition traits could potentially be used in crop improvement, if suitable markers can be identified and if these correspond with suitable agronomy and quality traits.


Cyclic Change of Fatty Acid Composition in Meibum During the Menstrual Cycle.
To evaluate the fatty acid (FA) composition in the meibum of pre- and postmenopausal women and age-matched men. This prospective study involved 24 healthy subjects; six premenopausal women in their 30s with a regular menstrual cycle (young-female [yF] group), six postmenopausal women in their 60s (elderly-female [eF] group), and 12 age-matched men (i.e., young-male [yM] and elderly-male [eM] groups, respectively). The menstrual cycle was divided into six phases (phase I-VI). Meibum was obtained from the meibomian gland orifices via a Daviel spoon, and its FA composition was then analyzed via gas chromatography mass spectrometry (GC-MS). Principal component analysis (PCA) was performed on the GC-MS results. The mean FA composition of all subjects was 40% saturated FAs (SFA) and 60% unsaturated FAs (UFAs). The PCA results of all groups indicated two categories (PC1 [77.5%] and PC2 [12.4%]); one consisting of yF-group samples of mainly phase II and III and the other consisting of the yF-group samples of the rest of the cycle, as well as from eF-group, yM-group, and eM-group samples. Each group had a distinctive nature. The FAs that most contributed to PC1 were C14:0, C16:0, and C18:0 in a positive correlation, and C18:1n9 in a negative correlation. FA composition noticeably changes during the menstrual cycle and is somewhat affected by sex and age. The ratio of SFAs (C16:0, C18:0) to mono-UFAs (C18:1n9) in the FA composition might have an impact on the lipid quality of meibum, thus suggesting alteration of its melting temperature and viscosity.


Heat Stress and Lipopolysaccharide Stimulation of Chicken Macrophage-Like Cell Line Activates Expression of Distinct Sets of Genes.
Acute heat stress requires immediate adjustment of the stressed individual to sudden changes of ambient temperatures. Chickens are particularly sensitive to heat stress due to development of insufficient physiological mechanisms to mitigate its effects. One of the symptoms of heat stress is endotoxemia that results from release of the lipopolysaccharide (LPS) from the guts. Heat-related cytotoxicity is mitigated by the innate immune system, which is comprised mostly of phagocytic cells such as monocytes and macrophages. The objective of this study was to analyze the molecular responses of the chicken macrophage-like HD11 cell line to combined heat stress and lipopolysaccharide treatment in vitro. The cells were heat-stressed and then allowed a temperature-recovery period, during which the gene expression was investigated. LPS was added to the cells to mimic the heat-stress-related endotoxemia. Semi high-throughput gene expression analysis was used to study a gene panel comprised of heat shock proteins, stress-related genes, signaling molecules and immune response genes. HD11 cell line responded to heat stress with increased mRNA abundance of the HSP25, HSPA2 and HSPH1 chaperones as well as DNAJA4 and DNAJB6 co-chaperones. The anti-apoptotic gene BAG3 was also highly up-regulated, providing evidence that the cells expressed pro-survival processes. The immune response of the HD11 cell line to LPS in the heat stress environment (up-regulation of CCL4, CCL5, IL1B, IL8 and iNOS) was higher than in thermoneutral conditions. However, the peak in the transcriptional regulation of the immune genes was after two hours of temperature-recovery. Therefore, we propose the potential influence of the extracellular heat shock proteins not only in mitigating effects of abiotic stress but also in triggering the higher level of the immune responses. Finally, use of correlation networks for the data analysis aided in discovering subtle differences in the gene expression (i.e. the role of the CASP3 and CASP9 genes).


Scalable exfoliation and dispersion of two-dimensional materials - an update.
The preparation of dispersions of single- and few-sheet 2D materials in various solvents, as well as the characterization methods applied to such dispersions, is critically reviewed. Motivating factors for producing single- and few-sheet dispersions of 2D materials in liquids are briefly discussed. Many practical applications are expected for such materials that do not require high purity formulations and tight control of donor and acceptor concentrations, as required in conventional Fab processing of semiconductor chips. Approaches and challenges encountered in exfoliating 2D materials in liquids are reviewed. Ultrasonication, mechanical shearing, and electrochemical processing approaches are discussed, and their respective limitations and promising features are critiqued. Supercritical and more conventional liquid and solvent processing are then discussed in detail. The effects of various types of stabilizers, including surfactants and other amphiphiles, as well as polymers, including homopolymeric electrolytes, nonionic polymers, and nanolatexes, are discussed. Consideration of apparent successes of stabilizer-free dispersions indicates that extensive exfoliation in the absence of dispersing aids results from processing-induced surface modifications that promote stabilization of 2D material/solvent interactions. Also apparent paradoxes in "pristineness" and optical extinctions in dispersions suggest that there is much we do not yet quantitatively understand about the surface chemistry of these materials. Another paradox, emanating from modeling dilute solvent-only exfoliation by sonication using polar components of solubility parameters and surface tension for pristine graphene with no polar structural component, is addressed. This apparent paradox appears to be resolved by realizing that the reactivity of graphene to addition reactions of solvent radicals produced by sonolysis is accompanied by unintended polar surface modifications that promote attractive interactions with solvent. This hypothesis serves to define important theoretical and experimental studies that are needed. We conclude that the greatest promise for high volume and high concentration processing lies in applying methods that have not yet been extensively reported, particularly wet comminution processing using small grinding media of various types.


Functional coassembly of KCNQ4 with KCNE-beta- subunits in Xenopus oocytes.
The KCNQ gene family comprises voltage-gated potassium channels expressed in epithelial tissues (KCNQ1, KCNQ5), inner ear structures (KCNQ1, KCNQ4) and the brain (KCNQ2-5). KCNQ4 is expressed in inner and outer hair cells of the inner ear where it influences electrical excitability and cell survival. Accordingly, loss of function mutations of the KCNQ4 gene cause hearing loss in humans and functional k.o.-mice show progressive degeneration of outer hair cells (OHCs). However, characteristic electrophysiological features of the native KCNQ4- carried current I(K,n) in OHCs are not recapitulated by expression of KCNQ4 channels in heterologous expression systems. This might suggest modulation of KCNQ4 by interacting KCNE Beta-subunits, which are known to modify the properties of the closely related KCNQ1. The present study explored whether transcripts of the KCNE isoforms could be identified in OHC mRNA and whether the subunits modulate KCNQ4 function. RT-PCR indeed yielded transcripts of all five KCNEs in OHCs. Coexpression of the KCNE- Beta-subunits with human KCNQ4 in the Xenopus laevis oocyte expression system revealed that all KCNEs modulate KCNQ4 voltage dependence, protein stability and ion selectivity of hKCNQ4 in Xenopus oocytes. The deafness-associated Jervell and Lange- Nielsen syndrome (JLNS) mutation KCNE1(D76N) impairs KCNQ4-function whereas the Romano-Ward syndrome (RWS) mutant KCNE1(S74L), which shows normal hearing in patients, does not impair KCNQ4 channel function. In conclusion, KCNEs are presumably coexpressed with KCNQ4 in hair cells from the organ of Corti and might regulate KCNQ4 functional properties, effects that could be important under physiological and pathophysiological conditions.


A nationwide randomized, double-blind, placebo-controlled physicians' trial of loxoprofen for the treatment of fatigue, headache, and nausea after hangovers.
Hangovers are associated with negative economic consequences due to decreased job performance or frequent visits to physicians. Thus, a new strategy for the alleviation of hangover-related symptoms is needed to avoid this detriment to society. The purpose of this nationwide randomized, double-blind, placebo-controlled physicians' trial was to evaluate the efficacy of loxoprofen sodium for the alleviation of fatigue, headache, and nausea after hangover. A total of 229 participants were randomized to receive loxoprofen sodium (60 mg once orally) or placebo. The study was closed when the first 150 participants (n = 74 in the loxoprofen vs. n = 76 in the placebo groups) experienced hangovers. The primary endpoint was set as the difference in severity of general fatigue before and 3 h after taking the test drugs and was evaluated using a visual analogue scale. Secondary endpoints included difference in severity of headache, nausea, and incidence of adverse events. The study participants were 34 (interquartile range; 30-39) years old, 92.0% were men, and both groups were comparable for baseline characteristics. The alleviation of general fatigue did not differ statistically between the loxoprofen and placebo groups (24 [14-49] vs. 19 [9-35], p = 0.07). However, the alleviation of headache was statistically greater in the loxoprofen group (25 [10-50] vs. 10 [2-30], adjusted difference 14, 95% confidence interval 8-21, p < 0.001), whereas, there was no difference in nausea (7 [0-27] vs. 10 [0-24], p = 0.68). The incidence of adverse symptoms such as epigastric discomfort was also comparable between groups (2.7% vs. 3.9%, p = 0.25). Loxoprofen sodium was effective for relieving headaches after hangovers but did not alleviate general fatigue or nausea.


Use of differential scanning fluorimetry as a high-throughput assay to identify nuclear receptor ligands.
Identification of ligands that interact with nuclear receptors is both a major biological problem and an important initial step in drug discovery. Several in vitro and in vivo techniques are commonly used to screen ligand candidates against nuclear receptors; however, none of the current assays allow screening without modification of either the protein and/or the ligand in a high-throughput fashion. Differential scanning fluorimetry (DSF) allows unmodified potential ligands to be screened as 10µL reactions in 96-well format against partially purified protein, revealing specific interactors. As a proof of principle, we used a commercially-available nuclear receptor ligand candidate chemical library to identify interactors of the human estrogen receptor α ligand binding domain (ERα LBD). Compounds that interact specifically with ERα LBD stabilize the protein and result in an elevation of the thermal denaturation point, as monitored by the environmentally-sensitive dye SYPRO orange. We successfully identified all three compounds in the library that have previously been identified to interact with ERα, with no false positive results.


Differential regulation of cystic fibrosis transmembrane conductance regulator and Na+,K+ -ATPase in gills of striped bass, Morone saxatilis: effect of salinity and hormones.
Effects of salinity and hormones on cystic fibrosis transmembrane conductance regulator (CFTR) and alpha-subunit Na(+),K(+) -ATPase (alpha-NKA) mRNA (analysed by semi-quantitative PCR) and protein expression (analysed by western blotting and immunocytochemistry) were investigated in gills of striped bass. Freshwater (FW) to seawater (SW) transfer induced a disturbance in serum [Na(+)]. Gill CFTR protein, mRNA level and Na(+),K(+) -ATPase activity were unaffected by SW transfer, whereas alpha-NKA mRNA increased after transfer. CFTR immunoreactivity was observed in large cells in FW and SW gill filaments at equal intensity. Cortisol decreased serum [Na(+)] in FW fish, but had no effect on gill Na(+),K(+) -ATPase activity, alpha-NKA and CFTR mRNA levels. Incubation of gill tissue with cortisol (24 h, >0.01 micro g/ml) and epidermal growth factor (EGF 10 micro g/ml) decreased CFTR mRNA levels relative to pre-incubation and control levels. CFTR expression was unaffected by IGF-I (10 micro g/ml). alpha-NKA mRNA levels decreased by 50% after 24 h control incubation; it was slightly stimulated by cortisol and unaffected by IGF-I and EGF. In isolated gill cells, phosphorylation of extracellular-regulated kinase (ERK) 1/2 was stimulated by EGF but not affected by IGF-I. This study is the first to report a branchial EGF response and to demonstrate a functional ERK 1/2 pathway in the teleost gill. In conclusion, CFTR and Na(+),K(+) -ATPase are differentially regulated by salinity and hormones in gills of striped bass, despite the putative involvement of both in salt excretion.


MicroRNAs reprogram normal fibroblasts into cancer-associated fibroblasts in ovarian cancer.
Cancer-associated fibroblasts (CAF) are a major constituent of the tumor stroma, but little is known about how cancer cells transform normal fibroblasts into CAFs. microRNAs (miRNA) are small noncoding RNA molecules that negatively regulate gene expression at a posttranscriptional level. Although it is clearly established that miRNAs are deregulated in human cancers, it is not known whether miRNA expression in resident fibroblasts is affected by their interaction with cancer cells. We found that in ovarian CAFs, miR-31 and miR-214 were downregulated, whereas miR-155 was upregulated when compared with normal or tumor-adjacent fibroblasts. Mimicking this deregulation by transfecting miRNAs and miRNA inhibitors induced a functional conversion of normal fibroblasts into CAFs, and the reverse experiment resulted in the reversion of CAFs into normal fibroblasts. The miRNA-reprogrammed normal fibroblasts and patient-derived CAFs shared a large number of upregulated genes highly enriched in chemokines, which are known to be important for CAF function. The most highly upregulated chemokine, CCL5, (C-C motif ligand 5) was found to be a direct target of miR-214. These results indicate that ovarian cancer cells reprogram fibroblasts to become CAFs through the action of miRNAs. Targeting these miRNAs in stromal cells could have therapeutic benefit. The mechanism by which quiescent fibroblasts are converted into CAFs is unclear. The present study identifies a set of 3 miRNAs that reprogram normal fibroblasts to CAFs. These miRNAs may represent novel therapeutic targets in the tumor microenvironment.


Three-dimensional microsurgical and tractographic anatomy of the white matter of the human brain.
We sought to investigate the three-dimensional structure of the white matter of the brain by means of the fiber-dissection technique and diffusion-tensor magnetic resonance imaging to assess the usefulness of the combination of both techniques, compare their results, and review the potential functional role of fiber tracts. Fifteen formalin-fixed human hemispheres were dissected according to Klingler's fiber-dissection technique with the aid of 36 to 340 magnification. Three-dimensional anatomic images were created with the use of specific software. Two hundred patients with neurological symptoms and five healthy volunteers were studied with diffusion-tensor magnetic resonance imaging (3 T) and tractographic reconstruction. The most important association, projection, and commissural fasciculi were identified anatomically and radiologically. Analysis of their localization, configuration, and trajectory was enhanced by the combination of both techniques. Three-dimensional anatomic reconstructions provided a better perception of the spatial relationships among the white matter tracts. Tractographic reconstructions allowed for inspection of the relationships between the tracts as well as between the tracts and the intracerebral lesions. The combination of topographical anatomic studies of human fiber tracts and neuroanatomic research in experimental animals, with data from the clinicoradiological analysis of human white matter lesions and intraoperative subcortical stimulation, aided in establishing the potential functional role of the tracts. The fiber-dissection and diffusion-tensor magnetic resonance imaging techniques are reciprocally enriched not only in their application to the study of the complex intrinsic architecture of the brain, but also in their practical use for diagnosis and surgical planning.


Preoperative risk factors for postoperative delirium following hip fracture repair: a systematic review.
Systematically identify preoperative clinical risk factors for incident postoperative delirium in individuals undergoing hip fracture repair in order to guide clinicians in identifying high risk patients at admission. This is a systematic review of prospective observational studies with estimation of association between preoperative risk factors and incident postoperative delirium in multivariate models. Electronic searches were conducted in PubMed, Embase, PsycINFO, CINAHL, Cochrane Library, Proquest Dissertations and Theses, and WorldCatDissertations. Hand searches were conducted in selected journals and their supplements. Search yielded 6380 titles and abstracts from electronic databases and 72 titles from hand searches, and 10 studies met inclusion criteria. The following risk factors were significant in bivariate models: cognitive impairment, age, gender, institutionalization, functional impairment, body mass index (BMI), albumin, comorbidities, American Society of Anesthesiologist classification, acute medical conditions, polypharmacy, and vision impairment. Among all of these risk factors, cognitive impairment most consistently remained statistically significant after adjusting for other risk factors in multivariate models, followed by BMI/albumin and multiple comorbidities. In our systematic review, cognitive impairment was one of the strongest preoperative risk factors for postoperative delirium after hip fracture surgery. Preoperative cognitive assessment may be one of the most useful methods of identifying those who are at high risk for postoperative delirium and prioritizing delivery of delirium prevention measures.


Midinfrared frequency combs from coherent supercontinuum in chalcogenide and optical parametric oscillation.
We observe the coherence of the supercontinuum generated in a nanospike chalcogenide-silica hybrid waveguide pumped at 2 μm. The supercontinuum is shown to be coherent with the pump by interfering it with a doubly resonant optical parametric oscillator (OPO) that is itself coherent with the shared pump laser. This enables coherent locking of the OPO to the optically referenced pump frequency comb, resulting in a composite frequency comb with wavelengths from 1 to 6 μm.


Distribution and phenotype of Phox2a-containing neurons in the adult sprague-dawley rat.
Phox2a is a transcription factor that plays an essential role, with Phox2b, in the specification of the adrenergic and noradrenergic phenotype in developing brain. Localization of Phox2a in developing brainstem has demonstrated a high degree of correspondence between neurons expressing the transcription factor and those involved in the regulation of autonomic function. Although it is well established that the paralogous gene product Phox2b is widely expressed in adult brain, no study has mapped the distribution of Phox2a in the adult. The data reported here address that void. A well-characterized rabbit polyclonal antiserum was used for immunohistochemical localization of the transcription factor in adult rats. Sections through the rostrocaudal extent of brain were processed for dual immunocytochemical localization of Phox2a and catecholamine enzymes. Adjacent sections were used for dual localization of Phox2a and NADPH diaphorase, a marker of nitric oxide-containing neurons. The data demonstrate that Phox2a is present in all brainstem catecholamine neurons, in circumscribed populations of NADPH(+) neurons, and in a subset of neurons that influences sympathetic and parasympathetic outflow. In addition, Phox2a(+) neurons were observed within diencephalic and brainstem nuclei that regulate behavioral state. Considered with data demonstrating that Phox2a is part of the transcriptional complex that drives expression of dopamine-beta-hydroxylase and can also up-regulate expression of other genes, the data support the conclusion that Phox2a plays an important role in brainstem catecholamine neurotransmission and in the regulation of adaptive homeostatic functions in the adult nervous system.


Enteroaggregative Escherichia coli Subclinical Infection and Coinfections and Impaired Child Growth in the MAL-ED Cohort Study.
We evaluated the impact of subclinical enteroaggregative Escherichia coli (EAEC) infection alone and in combination with other pathogens in the first 6 months of life on child growth. Nondiarrheal samples from 1684 children across 8 Multisite Birth Cohort Study, Malnutrition and Enteric Diseases (MAL-ED) sites in Asia, Africa, and Latin America were tested monthly; more than 90% of children were followed-up twice weekly for the first 6 months of life. Children with subclinical EAEC infection did not show altered growth between enrollment and 6 months. Conversely, EAEC coinfection with any other pathogen was negatively associated with delta weight-for-length (P < 0.05) and weight-for-age (P > 0.05) z scores between 0 and 6 months. The presence of 2 or more pathogens without EAEC was not significantly associated with delta weight-for-length and weight-for-age. The most frequent EAEC coinfections included Campylobacter spp, heat-labile toxin-producing enterotoxigenic E coli, Cryptosporidium spp, and atypical enteropathogenic E coli. Myeloperoxidase levels were increased with EAEC coinfection (P < 0.05). EAEC pathogen codetection was associated with lower neopterin levels compared to those of no-pathogen control children (P < 0.05). Mothers of children with EAEC coinfections had lower levels of education, poorer hygiene and sanitation, lower socioeconomic status, and lower breast-feeding rates compared to mothers of children in whom no pathogen was detected (P < 0.05). These data emphasize the public health importance of subclinical EAEC infection in early infancy in association with other pathogens and the need for improved maternal and child care, hygiene, sanitation, and socioeconomic factors.


Vitamin E supplementation in chronically hemodialyzed patients - influence on blood hemoglobin and plasma (anti)oxidant status.
Background: Disturbed oxidant/antioxidant status is involved in pathogenesis of anemia in end stage renal disease. There is evidence that vitamin E supplementation can increase blood hemoglobin in chronically hemodialyzed patients. However, the interindividual variation in response to the supplementation has not been fully addressed. Methods: 24 chronically hemodialyzed patients were supplemented with vitamin E (400 IU/day) in a period of two months. They had already been treated with erythropoiesis stimulating agents (ESA) and iron on a long-term basis, which was continued during the study period. A group of 20 healthy volunteers served as control subjects. Complete blood count, general biochemistry assays, the redox status by total thiols, oxidative stress by reactive oxygen metabolites, antioxidant status by biological antioxidant potential, and vitamin E (α- and γ- tocopherol) were measured before the start of supplementation, one month and two months later. Results: Overall, the vitamin E supplementation did not cause an increase of blood hemoglobin, hematocrit or red blood cells. However, 50 % of the patients with basal blood hemoglobin below 12.0 g/dL (N = 10) responded to the supplementation with its continuous increase. In addition, vitamin E exhibited a slight prooxidant effect only in the subgroup of patients with basal blood hemoglobin of ≥ 12.0 g/dL, two months after the start of supplementation (decreased total thiols: 300 ± 31 vs. 277 ± 36 µmol/L, p < 0.05; increased reactive oxygen metabolites: 183 ± 140 vs. 287 ± 112 CARR U, p > 0.05; decreased biological antioxidant potential: 2278 ± 150 vs. 2171 ± 126 µEq/L, p < 0.025), which coincided with their significantly increased serum α-tocopherol concentrations in comparison to the patients with basal blood hemoglobin below 12.0 g/dL (41.3 ± 7.2 vs. 59.9 ± 19.2 µmol/L, p < 0.025). Conclusions: When treated with ESA and iron on a long-term basis, the response to the vitamin E supplementation in chronically hemodialyzed patients is largely dependent on their basal blood hemoglobin and serum vitamin E concentrations.


Reducing the time complexity of the derandomized evolution strategy with covariance matrix adaptation (CMA-ES).
This paper presents a novel evolutionary optimization strategy based on the derandomized evolution strategy with covariance matrix adaptation (CMA-ES). This new approach is intended to reduce the number of generations required for convergence to the optimum. Reducing the number of generations, i.e., the time complexity of the algorithm, is important if a large population size is desired: (1) to reduce the effect of noise; (2) to improve global search properties; and (3) to implement the algorithm on (highly) parallel machines. Our method results in a highly parallel algorithm which scales favorably with large numbers of processors. This is accomplished by efficiently incorporating the available information from a large population, thus significantly reducing the number of generations needed to adapt the covariance matrix. The original version of the CMA-ES was designed to reliably adapt the covariance matrix in small populations but it cannot exploit large populations efficiently. Our modifications scale up the efficiency to population sizes of up to 10n, where n is the problem dimension. This method has been applied to a large number of test problems, demonstrating that in many cases the CMA-ES can be advanced from quadratic to linear time complexity.


THE REAL McCOIL: A method for the concurrent estimation of the complexity of infection and SNP allele frequency for malaria parasites.
As many malaria-endemic countries move towards elimination of Plasmodium falciparum, the most virulent human malaria parasite, effective tools for monitoring malaria epidemiology are urgent priorities. P. falciparum population genetic approaches offer promising tools for understanding transmission and spread of the disease, but a high prevalence of multi-clone or polygenomic infections can render estimation of even the most basic parameters, such as allele frequencies, challenging. A previous method, COIL, was developed to estimate complexity of infection (COI) from single nucleotide polymorphism (SNP) data, but relies on monogenomic infections to estimate allele frequencies or requires external allele frequency data which may not available. Estimates limited to monogenomic infections may not be representative, however, and when the average COI is high, they can be difficult or impossible to obtain. Therefore, we developed THE REAL McCOIL, Turning HEterozygous SNP data into Robust Estimates of ALelle frequency, via Markov chain Monte Carlo, and Complexity Of Infection using Likelihood, to incorporate polygenomic samples and simultaneously estimate allele frequency and COI. This approach was tested via simulations then applied to SNP data from cross-sectional surveys performed in three Ugandan sites with varying malaria transmission. We show that THE REAL McCOIL consistently outperforms COIL on simulated data, particularly when most infections are polygenomic. Using field data we show that, unlike with COIL, we can distinguish epidemiologically relevant differences in COI between and within these sites. Surprisingly, for example, we estimated high average COI in a peri-urban subregion with lower transmission intensity, suggesting that many of these cases were imported from surrounding regions with higher transmission intensity. THE REAL McCOIL therefore provides a robust tool for understanding the molecular epidemiology of malaria across transmission settings.


Loss of Eph-receptor expression correlates with loss of cell adhesion and chondrogenic capacity in Hoxa13 mutant limbs.
Mesenchymal patterning is an active process whereby genetic commands coordinate cell adhesion, sorting and condensation, and thereby direct the formation of morphological structures. In mice that lack the Hoxa13 gene, the mesenchymal condensations that form the autopod skeletal elements are poorly resolved, resulting in missing digit, carpal and tarsal elements. In addition, mesenchymal and endothelial cell layers of the umbilical arteries (UAs) are disorganized, resulting in their stenosis and in embryonic death. To further investigate the role of Hoxa13 in these phenotypes, we generated a loss-of-function allele in which the GFP gene was targeted into the Hoxa13 locus. This allele allowed FACS isolation of mesenchymal cells from Hoxa13 heterozygous and mutant homozygous limb buds. Hoxa13(GFP) expressing mesenchymal cells from Hoxa13 mutant homozygous embryos are defective in forming chondrogenic condensations in vitro. Analysis of pro-adhesion molecules in the autopod of Hoxa13 mutants revealed a marked reduction in EphA7 expression in affected digits, as well as in micromass cell cultures prepared from mutant mesenchymal cells. Finally, antibody blocking of the EphA7 extracellular domain severely inhibits the capacity of Hoxa13(GFP) heterozygous cells to condense and form chondrogenic nodules in vitro, which is consistent with the hypothesis that reduction in EphA7 expression affects the capacity of Hoxa13(-/-) mesenchymal cells to form chondrogenic condensations in vivo and in vitro. EphA7 and EphA4 expression were also decreased in the mesenchymal and endothelial cells that form the umbilical arteries in Hoxa13 mutant homozygous embryos. These results suggest that an important role for Hoxa13 during limb and UA development is to regulate genes whose products are required for mesenchymal cell adhesion, sorting and boundary formation.


Isolation and identification of antifungal peptides from Bacillus BH072, a novel bacterium isolated from honey.
A bacterial strain BH072 isolated from a honey sample showed antifungal activity against mold. Based on morphological, biochemical, physiological tests, and analysis of 16S rDNA sequence, the strain was identified to be a new subspecies of Bacillus sp. It had a broad spectrum of antifungal activity against various mold, such as Aspergillus niger, Pythium, and Botrytis cinerea. Six pairs of antifungal genes primers were designed and synthesized, and ituA, hag, tasA genes were detected by PCR analysis. The remarkable antifungal activity could be associated with the co-production of these three peptides. One of them was purified by 30-40% ammonium sulfate precipitation, Sephadex G-75 gel filtration and anion exchange chromatography on D201 resin. The purified peptide was estimated to be 35.615 kDa and identified to be flagellin by micrOTOF-Q II. By using methanol extraction, another substance was isolated from fermentation liquor, and determined to be iturin with liquid chromatography-mass spectrometry (LC-MS) method. The third possible peptide encoded by tasA was not isolated in this study. The culture liquor displayed antifungal activity in a wide pH range (5.0-9.0) and at 40-100°C. The result of the present work suggested that Bacillus BH072 might be a bio-control bacterium of research value.


Superior mesenteric venous injuries: to ligate or to repair remains the question.
Superior mesenteric vein injuries are rare and incur high mortality. Given their low incidence, little data exist delineating indications for when to institute primary repair versus ligation. The purposes of this study are to review our institutional experience, to determine the additive effect on mortality of associated vascular injuries, to correlate mortality with the American Association for the Surgery of Trauma-Organ Injury Scale (AAST-OIS) for abdominal vascular injury and to examine and define the indications and outcomes for primary repair versus ligation. Retrospective 156 months study (January 1992 through December 2004) in a large Level I urban trauma center of all patients admitted with superior mesenteric vein injuries. Patients were stratified, according to surgical technique employed to deal with their injuries, into those undergoing primary repair versus ligation to determine outcomes and define the surgical indications of these methods. The main outcome measure was overall survival. Cases of survival were stratified according to surgical method: primary repair versus ligation. There were 51 patients with a mean Injury Severity Score of 25 +/- 12. Mechanism of injury was penetrating for 38 (76%), blunt for 13 (24%), and patients undergoing emergency department thoracotomy for 4 (8%). Surgical management was ligation for 30 (59%), primary repair for 16 (31%), and 5 (10%) patients were exsanguinated before repair. The overall survival rate was 24/50 (47%). The survival rate excluding patients undergoing emergency department thoracotomy was 51%. The survival rate excluding patients that sustained greater than 3 to 4 associated vessels injured was 65%. The survival rates of patients with superior mesenteric vein and superior mesenteric artery was 55% and superior mesenteric vein and portal vein (PV) was 40%. The survival rate of patients with isolated superior mesenteric vein injuries was 55%. Mortality stratified to AAST-OIS grade III, 44%; grade IV, 42%; and grade V, 42%. Survival rates stratified to method of management consisted of primary repair (60%) versus ligation (40%). SMV injuries are highly lethal. Multiple associated vessel injuries increase mortality. Mortality correlates well with the American Association for the Surgery of Trauma-Organ Injury Scale for abdominal vascular injuries. Patients undergoing primary repair have higher survival rates (63%) and lesser numbers of associated vascular and nonvascular injuries; whereas those undergoing ligation have a smaller survival rate (40%) and higher number of associated vascular and nonvascular injuries. Ligation appears to be safe and should be selected for hemodynamically unstable patients with a large number of associated injuries.


Clinical benefit of metaphase I oocytes.
We studied the benefit of using in vitro matured metaphase I (MI) oocytes for ICSI in patients with a maximum of 6 mature metaphase II (MII) oocytes at retrieval. In 2004, 187 ICSI cycles were selected in which maximum 6 MII oocytes and at least one MI oocyte were retrieved. MI oocytes were put in culture to mature until the moment of ICSI, which was performed between 2 to 11 hours after oocyte retrieval (day 0). In exceptional cases, when the patient did not have any mature oocyte at the scheduled time of ICSI, MI oocytes were left to mature overnight and were injected between 19 to 26 hours after retrieval (day 1). Embryos from MI oocytes were chosen for transfer only when no other good quality embryos from MII oocytes were available. Outcome parameters were time period of in vitro maturation (IVM), IVM and fertilization rates, embryo development, clinical pregnancy rates, implantation rates and total MI oocyte utilization rate. The overall IVM rate was 43%. IVM oocytes had lower fertilization rates compared to in vivo matured sibling oocytes (52% versus 68%, P < 0.05). The proportion of poor quality embryos was significantly higher in IVM derived oocytes. One pregnancy and live birth was obtained out of 13 transfers of embryos exclusively derived from IVM oocytes. This baby originated from an oocyte that was injected after 22 hrs of IVM. Fertilization of in vitro matured MI oocytes can result in normal embryos and pregnancy, making IVM worthwhile, particularly when few MII oocytes are obtained at retrieval.


Characterization of bovine herpesvirus 1 UL49 homolog gene and product: bovine herpesvirus 1 UL49 homolog is dispensable for virus growth.
The sequence of the bovine herpesvirus 1 (BHV-1) gene that is homologous to the herpes simplex virus UL49 gene was determined. The BHV-1 UL49 homolog open reading frame consists of 774 bp and is capable of encoding 258 amino acids. Northern (RNA) blot analysis showed that the BHV-1 UL49 homolog is transcribed into a 1.1-kb RNA which is coterminal with the transcripts of an upstream UL49.5 homolog gene. Rabbit antisera produced against synthetic peptides of the predicted UL49 homolog gene product recognized a polypeptide of 33 to 35 kDa in both virus-infected cells and isolated virions. Further analysis by unionic-detergent partition of isolated virions suggested that the UL49 homolog gene product is a virion tegument protein. Indirect immunofluorescence assay revealed that the UL49 homolog gene product was predominantly localized in the nuclei of BHV-1-infected cells. A mutant virus with the UL49 homolog gene deleted was produced, and it was able to replicate in noncomplementing cells. Nevertheless, the yield of mutant virus was significantly reduced. The results from this study suggest that the BHV-1 UL49 homolog gene encodes a nuclear protein which constitutes a tegument component in mature virions and that it is dispensable for virus growth in cell culture.


Identification of the citrate exporter Cex1 of Yarrowia lipolytica.
Yarrowia lipolytica is a yeast with many talents, one of them being the production of citric acid. Although the citrate biosynthesis is well studied, little is known about the transport mechanism by which citrate is exported. To gain better insight into this mechanism, we set out to identify a transporter involved in citrate export of Y. lipolytica. A total of five proteins were selected for analysis based on their similarity to a known citrate exporter, but neither a citrate transport activity nor any other phenotypic function could be attributed to them. Differential gene expression analysis of two strains with a distinct citrate productivity revealed another three putative transporters, one of which is YALI0D20196p. Disrupting YALI0D20196g in Y. lipolytica abolished citrate production, while extrachromosomal expression enhanced citrate production 5.2-fold in a low producing wildtype. Furthermore, heterologous expression of YALI0D20196p in the non-citrate secreting yeast Saccharomyces cerevisiae facilitated citrate export. Likewise, expression of YALI0D20196p complemented the ability to secrete citrate in an export-deficient strain of Aspergillus niger, confirming a citrate export function of YALI0D20196p. This report on the identification of the first citrate exporter in Y. lipolytica, termed Cex1, represents a valuable starting point for further investigations of the complex transport processes in yeasts.


Rates of prescription opiate use before and after injury in patients with orthopaedic trauma and the risk factors for prolonged opiate use.
The prudent use of prescription opiate medications is a central aspect of postoperative pain management. The mortality associated with prescription opiate overdose is reaching epidemic proportions nationally, and is the leading cause of accidental death in greater than half the states in the United States. This study sought to determine the rates of preinjury opiate use in patients with orthopaedic trauma and the risk factors for prolonged use postinjury. The Utah Controlled Substance Database was queried to determine the use of prescription opiates by all patients admitted to the orthopaedic trauma service for a two-year period with isolated musculoskeletal injuries. Usage three months prior to injury and six months postinjury was examined. Six hundred and thirteen patients met inclusion criteria. Among patients with orthopaedic trauma, 15.5% had filled a prescription for opiates in the three months prior to injury, compared with 9.2% of the general population (p < 0.001). More than one prescription was filled by 12.2% of the patients with trauma preinjury compared with 6.4% of the general population (p < 0.001). Postoperatively, 68.4% of all patients filled opiate prescriptions for less than six weeks, 11.9% filled opiate prescriptions between six and twelve weeks, and 19.7% filled opiate prescriptions past twelve weeks. Patients with preinjury use of more than one opiate prescription in the three months preinjury were six times as likely to continue use past twelve weeks, and 3.5 times as likely to obtain opiates from a provider other than their surgeon (p < 0.001). Opiate use was briefest with upper-extremity injuries, followed by lower-extremity injuries and pelvic or acetabular injuries. Regression models demonstrate that risk factors for prolonged use of opiates include advancing age and extent of preinjury use. Patients with orthopaedic trauma are significantly more likely than the general population to use prescription opiates prior to injury. Preinjury opiate use is predictive of prolonged use postinjury and predictive of patients who will seek opiates from other providers.


Boucher-Neuhäuser syndrome: cerebellar degeneration, chorioretinal dystrophy and hypogonadotropic hypogonadism: two novel cases and a review of 40 cases from the literature.
The combination of progressive cerebellar degeneration, hypogonadotropic hypogonadism and chorioretinal dystrophy defines the rare Boucher-Neuhäuser syndrome (BNS), which has recently been linked to autosomal-recessive mutations in the PNPLA6 gene in four index patients. Here we present two novel unrelated patients with BNS, where we identified four recessive PNPLA6 mutations (3 of them novel) as the genetic cause, using a targeted high-throughput approach. This finding provides the first replication from independent families that BNS is caused by PNPLA6 and, moreover, highlights PNPLA6 as the major gene leading to BNS. Given the fact that the major gene causing BNS has thus now been identified, we summarize the spectrum of clinical presentations and phenotype evolution of BNS based on a systematic in-depth review of the literature of previously published cases (n = 40). Both the two cases presented here and our review of the literature propose that the clinical presentation of BNS can be variable regarding both the age (ranging from 1 to 40 years) and the clinical symptoms at onset (cerebellar ataxia in 38 %; vision loss in 36 %; delayed puberty in 26 %). A substantial fraction of BNS cases may present with relatively selective atrophy of the superior and dorsal parts of the cerebellar vermis along with atrophy of the cerebellar hemispheres on MRI, while brainstem or cortical changes on MRI seem to be present only in small fractions. Also in the literature, no other major genetic causes of BNS other than PNPLA6 mutations were identified.


Screening for submicroscopic chromosome rearrangements in children with idiopathic mental retardation using microsatellite markers for the chromosome telomeres.
Recently much attention has been given to the detection of submicroscopic chromosome rearrangements in patients with idiopathic mental retardation. We have screened 27 subjects with mental retardation and dysmorphic features for such rearrangements using a genetic marker panel screening. The screening was a pilot project using markers from the subtelomeric regions of all 41 chromosome arms. The markers were informative for monosomy in both parents at 3661902 loci (40.6%, 95% confidence interval 37.0-44.2%) in the 22 families where DNA was available from both parents. In two of the 27 subjects, submicroscopic chromosomal aberrations were detected. The first patient had a 5-6 Mb deletion of chromosome 18q and the second patient had a 4 Mb deletion of chromosome 1p. The identification of two deletions in 27 cases gave an aberration frequency of 7.5% without adjustment for marker informativeness (95% confidence interval 1-24%) and an estimated frequency of 18% if marker informativeness for monosomy was taken into account. This frequency is higher than previous estimates of the number of subtelomeric chromosome abnormalities in children with idiopathic mental retardation (5-10%) although the confidence interval is overlapping. Our study suggests that in spite of the low informativeness of this pilot screening, submicroscopic chromosome aberrations may be a common cause of dysmorphic features and mental retardation.


Effects of prolonged expiration breathing on cardiopulmonary responses during incremental exercise.
This study was designed to clarify the effects of breathing with prolonged expiration on cardiopulmonary responses and autonomic nervous activity during incremental exercise. Eleven healthy men were randomly assigned to breathing mode: a prolonged expiration breathing with a 2-s inspired time and 4-s expired time and a spontaneous breathing without any constraints. Oxygen uptake (V(O2)), ventilation efficiency (V(E)/V(CO2)) and rate pressure product were measured. Low- (LF) and high-frequency (HF) components of blood pressure and heart rate variability were analyzed to assess sympathetic and parasympathetic nervous activities, respectively. V(E)/V(CO2), rate pressure product and LF were significantly lower, and [Formula: see text] and HF were significantly higher during exercise with prolonged expiration than with spontaneous breathing. Striking effects of prolonged expiration breathing included the improvement of ventilation efficiency, the suppression of sympathetic nervous activity and the activation of parasympathetic one during incremental exercise. Furthermore, prolonged expiration breathing may have suppressed the exercise-induced increase in myocardial V(O2).


Phytochemical composition and larvicidal activity of essential oils from herbal plants.
The essential oils (EOs) of Plectranthus amboinicus showed the highest larvicidal activity among four herbal plants studied and β-caryophyllene might be the major component responsible for its differential toxicity to the larvae of Culex quinquefasciatus and Aedes Aegypti. Mosquitoes act as vectors for many life-threatening diseases, including malaria, dengue fever, and Zika virus infection. Management of mosquitoes mainly relies on synthetic insecticides, which usually result in the rapid development of resistance; therefore, alternative mosquito control strategies are urgently needed. This study characterized the major component of essential oils (EOs) derived from the vegetative parts of four herbal plants and their larvicidal activity toward important mosquito vectors. The EOs were extracted by hydro-distillation and subjected to gas chromatography-mass spectrometry (GC-MS) analysis and a larvicidal activity assay toward Aedes aegypti, Ae. albopictus and Culex quinquefasciatus. In total, 14, 11, 11 and 9 compounds were identified from the EOs of Plectranthus amboinicus, Mentha requienii, Vitex rotundifolia and Crossostephium chinense, respectively. The EOs derived from four herbal plants exhibited remarkable larvicidal activity against the three mosquito species. In particular, the EOs of P. amboinicus showed the highest larvicidal activity, and the larvae of Cx. quinquefasciatus were more sensitive to the P. amboinicus EOs than that of Ae. Aegypti. Although carvacrol (61.53%) was the predominant constituent of the P. amboinicus EOs, its precursors, γ-terpinene (8.51%) and p-cymene (9.42%), exhibited the most larvicidal activity toward Ae. aegypti and Cx. quinquefasciatus. However, β-caryophyllene (12.79%) might be the major component responsible for the differential toxicity of the P. amboinicus EOs, as indicated by the significant differences in its LC50 values toward both mosquitoes. Information from these studies will benefit the incorporation of EOs into integrated vector management.