Consequences of parental burnout: Its specific effect on child neglect and violence.
Parental burnout is a specific syndrome resulting from enduring exposure to chronic parenting stress. It encompasses three dimensions: an overwhelming exhaustion related to one's parental role, an emotional distancing from one's children and a sense of ineffectiveness in one's parental role. This study aims to facilitate further identification of the consequences of parental burnout for the parents themselves, their spouses and their child(ren). In a sample of 1551 parents, we examined the relationship between parental burnout and seven possible consequences: escapism and suicidal thoughts, addictions, sleep disorders, marital conflicts, a partner estrangement mindset, and neglect and violence towards one's child(ren). We examined (1) to what extent parental and job burnout related to each of these possible consequences and (2) whether parental burnout is specifically related to neglectful and violent behaviour towards one's child(ren). The results suggest that parental burnout has a statistically similar effect to job burnout on addictions and sleep problems, a stronger effect on couples' conflicts and partner estrangement mindset and a specific effect on child-related outcomes (neglect and violence) and escape and suicidal ideation. These results emphasize the importance of accurately diagnosing this syndrome.


Everything You Always Wanted to Know About Salmonella Type 1 Fimbriae, but Were Afraid to Ask.
Initial attachment to host intestinal mucosa after oral infection is one of the most important stages during bacterial pathogenesis. Adhesive structures, widely present on the bacterial surface, are mainly responsible for the first contact with host cells and of host-pathogen interactions. Among dozens of different bacterial adhesins, type 1 fimbriae (T1F) are one of the most common adhesive organelles in the members of the Enterobacteriaceae family, including Salmonella spp., and are important virulence factors. Those long, thin structures, composed mainly of FimA proteins, are responsible for recognizing and binding high-mannose oligosaccharides, which are carried by various glycoproteins and expressed at the host cell surface, via FimH adhesin, which is presented at the top of T1F. In this review, we discuss investigations into the functions of T1F, from the earliest work published in 1958 to operon organization, organelle structure, T1F biogenesis, and the various functions of T1F in Salmonella-host interactions. We give special attention to regulation of T1F expression and their role in binding of Salmonella to cells, cell lines, organ explants, and other surfaces with emphasis on biofilm formation and discuss T1F role as virulence factors based on work using animal models. We also discuss the importance of allelic variation in fimH to Salmonella pathogenesis, as well as role of FimH in Salmonella host specificity.


Twelve tips for evaluating educational programs.
At one time or another, nearly all educators will need to evaluate an educational program to determine its merit or worth. These tips will help readers collect information to inform a meaningful evaluation, whether for local use or broad dissemination (i.e., research). The two most important questions in any evaluation are, 'Whose opinion matters?' and 'What would really be meaningful to them?' Other key steps include getting input from others, focusing on desired outcomes before selecting instruments, considering the validity or trustworthiness of the data, and pilot testing the evaluation process.


Measuring the quality of preventive and developmental services for young children: national estimates and patterns of clinicians' performance.
To generate a national picture of performance in the area of preventive and developmental services for children aged 4 to 35 months using 4 composite quality measures in the areas of 1) anticipatory guidance and parental education, 2) screening for family psychosocial risks, 3) screening for smoking and drug and alcohol use in the home, and 4) provision of family-centered care. Data from the National Survey on Early Childhood Health (N = 2068) were used to calculate the 4 composite performance measures, which, taken together, represent 23 topics included in the American Academy of Pediatrics health supervision guidelines. The reliability and degree of redundancy within and across these 4 measures were evaluated. Four methods for scoring these measures were used. Quality scores for subgroups of children were calculated, and logistic regression analysis was performed to examine the association of demographic, health, and health system variables with receiving recommended care. Regardless of the scoring method used, performance is highest in areas of family-centered care and screening for smoking and drug and alcohol use in the home. Performance is lowest in the areas of anticipatory guidance and education and assessment for family psychosocial risks. Using a scoring method that takes into account parent preferences for guidance and beliefs about discussing psychosocial topics, composite quality measure scores ranged from 13.5% to 59.6% of parents of young children receiving recommended care. Overall, 94.0% of parents reported 1 or more unmet needs for parenting guidance, education, and screening by pediatric clinician(s) in 1 or more of the content of care areas evaluated. Uninsured children and children aged 18 to 35 months are disproportionately represented among the 15.3% of children whose parents indicated an unmet need in each of the 4 areas of care. Although the reliability of each composite measure was high, no single item in any composite was highly correlated with the remaining combined items. Performance on any 1 composite measure for a child was only somewhat predictive of performance for the other measures. There are significant variations in performance on the basis of child age, race, insurance status, maternal education, marital status, and parent language as well as other factors. National results using 4 complementary composite quality measures confirm the need for improving the quality of preventive and developmental services for young children in the United States. The 4 measures identify areas of care and subgroups of children for whom improvements in quality are most needed. The measures provide a parsimonious yet comprehensive assessment across distinct health supervision topics and 4 essential aspects of preventive and developmental services. Until valid measures of outcomes of preventive and developmental services are identified or 1 single process of care measure is shown to be highly predictive of these outcomes, assessing multiple aspects of recommended care will be necessary to assess performance of health care providers or systems of care.


Development of a Novel Cell Surface Attachment System to Display Multi-Protein Complex Using the Cohesin-Dockerin Binding Pair.
Autodisplay of a multimeric protein complex on a cell surface is limited by intrinsic factors such as the types and orientations of anchor modules. Moreover, improper folding of proteins to be displayed often hinders functional cell surface display. While overcoming these drawbacks, we ultimately extended the applicability of the autodisplay platform to the display of a protein complex. We designed and constructed a cell surface attachment (CSA) system that uses a noncovalent protein-protein interaction. We employed the high-affinity interaction mediated by an orthogonal cohesin-dockerin (Coh-Doc) pair from Archaeoglobus fulgidus to build the CSA system. Then, we validated the orthogonal Coh-Doc binding by attaching a monomeric red fluorescent protein to the cell surface. In addition, we evaluated the functional anchoring of proteins fused with the Doc module to the autodisplayed Coh module on the surface of Escherichia coli. The designed CSA system was applied to create a functional attachment of dimeric α-neoagarobiose hydrolase to the surface of E. coli cells.


Diabetes and risk of Parkinson's disease.
To investigate the relationship between diabetes and future risk of Parkinson's disease (PD) among older U.S. adults. A prospective study of self-reported diabetes in 1995 and 1996 in relation to PD diagnosed after 1995 among 288,662 participants of the National Institutes of Health-AARP Diet and Health Study. Multivariate odds ratio (OR) and 95% CI were derived from logistic regression models. A total of 1,565 participants with PD diagnosed after 1995 were included in the analysis. After adjustment for potential confounders, PD risk was ∼40% higher (OR = 1.41 [95% CI 1.20-1.66]) among diabetic patients than among participants without diabetes. Further analysis showed that the risk elevation was largely limited to individuals who had diabetes for more than 10 years at the time of baseline survey (1.75 [1.36-2.25]). The association with diabetes was seen for both participants with PD diagnosed between 1995 and 1999 and participants with PD diagnosed after 2000. In addition, similar results were obtained after excluding participants with stroke, heart disease, cancers, or poor or fair health status and in subgroup analyses by age, sex, smoking status, and coffee consumption. This large study showed that diabetes was associated with a higher future risk of PD and the nature of this association warrants further investigation.


Whole-genome gene expression modifications associated with nitrosamine exposure and micronucleus frequency in human blood cells.
N-nitroso compounds (NOCs) are suspected human carcinogens and relevant in human exposure. NOCs also induce micronuclei (MN) formation in vivo. Since lymphocytic MN represent a validated biomarker of human cancer risk, establishing a link between NOC exposure and MN frequency in humans and concurrently investigating associated transcriptomic responses may provide crucial information on underlying molecular mechanisms that predispose to carcinogenicity. We used lymphocytes, from adult females participating in the pan-European biomarker research project NewGeneris, as a surrogate tissue for analysing such potentially carcinogenic gene expression and MN formation events in target organs. To assess NOC exposure, urine samples were analysed for marker nitrosamines. NOC excretion levels and MN frequency were subsequently linked to peripheral blood transcriptomics. We demonstrated a significant association between MN frequency and urinary NOCs (r = 0.41, P = 0.025) and identified modifications in among others cytoskeleton remodeling, cell cycle, apoptosis and survival, signal transduction, immune response, G-protein signaling and development pathways, which indicate a response to NOC-induced genotoxicity. Moreover, we established a network of genes, the most important ones of which include FBXW7, BUB3, Caspase 2, Caspase 8, SMAD3, Huntingtin and MGMT, which are involved in processes relevant in carcinogenesis. The modified genetic processes and genes found in this study may be of interest for future investigations into the potential carcinogenic risk associated with NOC exposure in humans.


A Decision Tree-Initialised Neuro-fuzzy Approach for Clinical Decision Support.
Apart from the need for superior accuracy, healthcare applications of intelligent systems also demand the deployment of interpretable machine learning models which allow clinicians to interrogate and validate extracted medical knowledge. Fuzzy rule-based models are generally considered interpretable that are able to reflect the associations between medical conditions and associated symptoms, through the use of linguistic if-then statements. Systems built on top of fuzzy sets are of particular appealing to medical applications since they enable the tolerance of vague and imprecise concepts that are often embedded in medical entities such as symptom description and test results. They facilitate an approximate reasoning framework which mimics human reasoning and supports the linguistic delivery of medical expertise often expressed in statements such as 'weight low' or 'glucose level high' while describing symptoms. This paper proposes an approach by performing data-driven learning of accurate and interpretable fuzzy rule bases for clinical decision support. The approach starts with the generation of a crisp rule base through a decision tree learning mechanism, capable of capturing simple rule structures. The crisp rule base is then transformed into a fuzzy rule base, which forms the input to the framework of adaptive network-based fuzzy inference system (ANFIS), thereby further optimising the parameters of both rule antecedents and consequents. Experimental studies on popular medical data benchmarks demonstrate that the proposed work is able to learn compact rule bases involving simple rule antecedents, with statistically better or comparable performance to those achieved by state-of-the-art fuzzy classifiers.


Inhibition of Wnt/β-catenin signaling suppresses bleomycin-induced pulmonary fibrosis by attenuating the expression of TGF-β1 and FGF-2.
Pulmonary fibrosis is a progressive lung disorder of unknown etiology, which is characterized by alterations in alveolar epithelium function, fibroblast activation, and increased extracellular matrix deposition. Recent studies have demonstrated that PF is associated with uncontrolled production of cytokines after lung injury. In the present study, we found that transforming growth factor-β1 (TGF-β1) and fibroblast growth factor 2 (FGF-2) were both upregulated in bleomycin-induced fibrotic lung tissue and primary murine alveolar epithelial Type II (ATII) cells treated with bleomycin. Furthermore, we discovered that TGF-β1 could induce the differentiation of lung resident mesenchymal stem cells (LR-MSCs) into fibroblasts, which may play an essential role in PF. LR-MSCs incubated with FGF-2 showed modest alterations in the expression of α-SMA and Vimentin. Moreover, in our study, we found that Wnt/β-catenin signaling was activated both in vitro and in vivo as a result of bleomycin treatment. Interestingly, we also found that suppression of the Wnt/β-catenin signaling could significantly attenuate bleomycin-induced PF accompanied with decreased expression of TGF-β1 and FGF-2 in vitro and in vivo. These results support that controlling the aberrant expression of TGF-β1 and FGF-2 via inhibition of Wnt/β-catenin signaling could serve as a potential therapeutic strategy for PF.


In Vitro Activity of Isavuconazole against Opportunistic Fungal Pathogens from Two Mycology Reference Laboratories.
Monitoring antifungal susceptibility patterns for new and established antifungal agents seems prudent given the increasing prevalence of uncommon species associated with higher antifungal resistance. We evaluated the activity of isavuconazole against 4,856 invasive yeasts and molds collected worldwide. The 4,856 clinical fungal isolates, including 2,351 Candida species isolates, 97 non-Candida yeasts, 1,972 Aspergillus species isolates, and 361 non-Aspergillus molds, including 292 Mucorales isolates collected in 2015 to 2016, were tested using CLSI methods. The MIC values for isavuconazole versus Aspergillus ranged from 0.06 to ≥16 μg/ml. The modal MIC for isavuconazole was 0.5 μg/ml (range, 0.25 [A. nidulans and A. terreus species complex] to 4 μg/ml [A. calidoustus and A. tubingensis]). Eight A. fumigatus isolates had elevated isavuconazole MIC values at ≥8 μg/ml (non-wild type). Isavuconazole showed comparable activity to itraconazole against the Mucorales The lowest modal isavuconazole MIC values were seen for Rhizopus spp., R. arrhizus var. arrhizus, and R. microsporus (all 1 μg/ml). Candida species isolates were inhibited by ≤0.25 μg/ml of isavuconazole (range, 96.1% [C. lusitaniae] to 100.0% [C. albicans, C. dubliniensis, C. kefyr, and C. orthopsilosis]). MIC values were ≤1 μg/ml for 95.5% of C. glabrata isolates and 100.0% of C. krusei isolates. Isavuconazole was active against the non-Candida yeasts, including Cryptococcus neoformans (100.0% at ≤0.5 μg/ml). Isavuconazole exhibited excellent activity against most species of Candida and Aspergillus Isavuconazole was comparable to posaconazole and voriconazole against the less common yeasts and molds. Isavuconazole was generally less active than posaconazole and more active than voriconazole against the 292 Mucorales isolates. We confirm the potentially useful activity of isavuconazole against species of Rhizopus as determined by CLSI methods.


Glucose and non-glucidic nutrients exert permissive effects on 2-keto acid regulation of pancreatic beta-cell function.
Insulin-releasing effects of straight and branched chain 2-keto acids were assessed using clonal glucose-responsive beta-cells. Pyruvic acid (PA), 2-ketovaleric acid (KV), 2-ketoisovaleric acid (KIV) or 2-keto-3-methylvaleric acid (KMV) dose-dependently promoted the stimulatory effects of D-glucose, whereas 2-ketobutyric acid (KB) did not affect insulin release. The stimulatory 2-keto acids also promoted the stimulatory activity of D-glyceraldehyde, L-leucine or L-arginine. Responses to PA, KV, KIV or KMV were significantly reduced by transport inhibition with 2-cyano-3 hydroxycinnamate, glucokinase inhibition with mannoheptulose or metabolic inhibition with sodium azide or sodium cyanide. Membrane hyperpolarisation with K+ depletion or diazoxide reduced insulin output, but failed to abolish secretory responses to KV, KIV and KMV. Secretory effects of these 2-keto acids also persisted in beta-cells depolarised with high KCl and glucose. Voltage-dependent Ca2+ channel blockade, with verapamil, or depletion of extracellular Ca2+ abolished the secretory activity of 2-keto acids. Collectively, these results indicate that glucose and metabolisable nutrients exert permissive effects on 2-keto acid-induced insulin release. In addition, KV, KIV and KMV can regulate beta-cell function at least partially independently of K+-ATP channel activity, both through their mitochondrial metabolism and regulation of Ca2+ influx.


Ondansetron Exposure Changes in a Pregnant Woman.
Pregnancy results in many physiologic changes that can alter the pharmacokinetic profiles of medications used during pregnancy. One of the primary factors leading to these pharmacokinetic changes is altered activity of drug-metabolizing enzymes. Ondansetron is a substrate of cytochrome P450 (CYP) 3A4 (primary metabolic pathway), 2D6, and 1A2, all of which are altered during pregnancy. We evaluated the pharmacokinetics of ondansetron at three different gestational time points in a 26-year-old, pregnant, Caucasian woman with normal liver and kidney function, who was maintained on ondansetron 8 mg administered orally 3 times/day throughout her pregnancy. Serial plasma samples were collected from the subject over one 8-hour dosing interval at 14, 24, and 35 weeks' gestation (representing early-, mid-, and late-pregnancy time points, respectively). Ondansetron plasma concentrations were determined using liquid chromatography-tandem mass spectrometry. Ondansetron area under the plasma concentration-time curve decreased progressively across gestation (634 ng hr/ml in early pregnancy, 553 ng hr/ml in mid-pregnancy, and 387 ng hr/ml in late pregnancy), with a corresponding increase in apparent oral clearance (12.6 L/hr in early-pregnancy, 14.5 L/hr in mid-pregnancy, and 20.7 L/hr in late-pregnancy). The decreased area under the plasma concentration-time curve and exposure to ondansetron across gestation is likely due to increased activity of CYP3A4 and CYP2D6 during pregnancy. We were not able to study this patient during the postpartum period; however, as with other CYP3A4 and CYP2D6 substrates, the apparent activities of these isoenzymes are likely return to baseline. To our knowledge, this is the first report to describe ondansetron pharmacokinetics across gestation. Additional pharmacokinetic and pharmacodynamic data are needed to confirm our results and to evaluate clinical impact; however, in the meantime, clinicians should be aware of these pharmacokinetic changes in ondansetron exposure during pregnancy.


Octave-spanning coherent supercontinuum generation in an AlGaAs-on-insulator waveguide.
We demonstrate supercontinuum generation over an octave spaning from 1055 to 2155 nm on the highly nonlinear aluminum gallium arsenide (AlGaAs)-on-insulator platform. This is enabled by the generation of two dispersive waves in a 3-mm-long dispersion-engineered nano-waveguide. The waveguide is pumped at telecom wavelengths (1555 nm) with 3.6 pJ femtosecond pulses. We experimentally validate the coherence of the generated supercontinuum around the pump wavelength (1450-1750 nm), and our numerical simulation shows a high degree of coherence over the full spectrum.


Significant regional variation exists in morbidity and mortality after repair of abdominal aortic aneurysm.
Limited data exist comparing perioperative morbidity and mortality after open and endovascular abdominal aortic aneurysm (AAA) repair (EVAR) among regions of the United States. This study evaluated the regional variation in mortality and perioperative outcomes after repair of AAAs. The Vascular Quality Initiative (VQI) was used to identify patients undergoing open AAA repair and EVAR between 2009 and 2014. Ruptured and intact aneurysms were evaluated separately, and the analysis of intact aneurysms was limited to infrarenal AAAs. All 16 regions of the VQI were deidentified, and those with <100 open repairs were combined to eliminate the effect of low-volume regions. Regional variation was evaluated using χ2 and Fisher exact tests. Regional rates were compared against current quality benchmarks. Perioperative outcomes from 14 regions were compared. After open repair of intact aneurysms, no significant variation was seen in 30-day or in-hospital mortality; however, multiple regions exceeded the Society for Vascular Surgery benchmark for in-hospital mortality after open repair of intact aneurysms of <5% (range, 0%-7%; P = .55). After EVAR, all regions met the Society for Vascular Surgery benchmark of <3% (range, 0%-1%; P = .75). Significant variation in in-hospital mortality existed after open (14%-63%; P = .03) and endovascular (3%-32%; P = .03) repair of ruptured aneurysms across the VQI regional groups. After repair of intact aneurysms, wide variation was seen in prolonged length of stay (>7 days for open repair: 32%-53%, P = .54; >2 days for EVAR: 16-43%, P < .01), transfusion (open: 10%-35%, P < .01; EVAR: 7%-18%, P < .01), use of vasopressors (open: 19%-37%, P < .01; EVAR: 3%-7%, P < .01), and postoperative myocardial infarction (open: 0%-13%, P < .01; EVAR: 0%-3%, P < .01). After open repair, worsening renal function (6%-18%; P = .04) and respiratory complications (6%-20%; P = .20) were variable across regions. The frequency of endoleak at completion of EVAR also had considerable variation (15%-38%; P < .01). Despite limited variation, multiple regions do not meet current benchmarks for in-hospital mortality after open AAA repair for intact aneurysms. Significant regional variation exists in perioperative outcomes and length of stay, and mortality is widely variable after repair for rupture. These data identify important areas for quality improvement initiatives and clinical practice guidelines.


Natural whisker-guided behavior by head-fixed mice in tactile virtual reality.
During many natural behaviors the relevant sensory stimuli and motor outputs are difficult to quantify. Furthermore, the high dimensionality of the space of possible stimuli and movements compounds the problem of experimental control. Head fixation facilitates stimulus control and movement tracking, and can be combined with techniques for recording and manipulating neural activity. However, head-fixed mouse behaviors are typically trained through extensive instrumental conditioning. Here we present a whisker-based, tactile virtual reality system for head-fixed mice running on a spherical treadmill. Head-fixed mice displayed natural movements, including running and rhythmic whisking at 16 Hz. Whisking was centered on a set point that changed in concert with running so that more protracted whisking was correlated with faster running. During turning, whiskers moved in an asymmetric manner, with more retracted whisker positions in the turn direction and protracted whisker movements on the other side. Under some conditions, whisker movements were phase-coupled to strides. We simulated a virtual reality tactile corridor, consisting of two moveable walls controlled in a closed-loop by running speed and direction. Mice used their whiskers to track the walls of the winding corridor without training. Whisker curvature changes, which cause forces in the sensory follicles at the base of the whiskers, were tightly coupled to distance from the walls. Our behavioral system allows for precise control of sensorimotor variables during natural tactile navigation.


Can early liver biopsies predict long-term outcome of the graft?
Chronic rejection (CR) in liver allografts show a rapid onset and progressive course, leading to graft failure within the first year after transplantation. Most cases are preceded by episodes of acute cellular rejection (AR), but histological features predictive for the transition toward CR are not well documented. We assessed the predictive value of centrilobular necrosis, central vein endothelialitis (CVE), central vein fibrosis, and lobular inflammation in the development of CR. One-week and one-month biopsy specimens of 12 patients with CR were compared with those of a control group consisting of 17 patients, who experienced AR without developing CR. The progress of the histological changes was further evaluated in follow-up biopsy specimens of the CR group taken at 2 months and beyond 3 months after transplantation. Centrilobular necrosis, CVE, central vein fibrosis, and lobular inflammation were common features in both groups at 1 week. At 1 month, the incidence declined in the control group. The CR group showed an increased incidence and persistence of these features in the follow-up specimens. The incidence and median grade of severity of CVE was significantly higher in the CR group (p=0.04, and P<0.001). The severity of portal and lobular inflammation was also more pronounced in the CR group (P+0.01 and 0.069). Conversely, in the control group the incidence of the lobular features decreased and the severity of CVE declined significantly (P=0.03). The shift from a predominantly portal-based process toward lobular graft damage represents the early transition of AR to CR, for which a modification of immunosuppression might be necessary to prevent graft loss.


Major global radiation of corvoid birds originated in the proto-Papuan archipelago.
A central paradigm in island biogeography has been the unidirectional "downstream" colonization of islands from continents (source to sink) based on the idea that less-diverse island communities are easier to invade than biologically more-diverse continental communities. Recently, several cases of "upstream" colonization (from islands to continents) have been documented, challenging the traditional view. However, all these cases have involved individual island species that have colonized mainland regions. Here, using molecular phylogenetic data, divergence time estimates, lineage diversity distributions, and ancestral area analyses, we reconstruct the spread of a species-rich (>700 species) passerine bird radiation (core Corvoidea) from its late Eocene/Oligocene origin in the emerging proto-Papuan archipelago north of Australia, including multiple colonizations from the archipelago to Southeast Asia. Thus, islands apparently provided the setting for the initiation of a major songbird radiation that subsequently invaded all other continents. Morphological and behavioral adaptations of the core Corvoidea as generalist feeders in open habitats, which facilitated dispersal and colonization, apparently evolved in the descendants of sedentary forest birds that invaded the proto-Papuan archipelago. The archipelago evidently provided islands of the right size, number, and proximity to continental areas to support the adaptation and diversification of vagile colonizers that went on to increase avian diversity on a global scale.


Management of upper airway edema caused by hereditary angioedema.
Hereditary angioedema is a rare disorder with a genetic background involving mutations in the genes encoding C1-INH and of factor XII. Its etiology is unknown in a proportion of cases. Recurrent edema formation may involve the subcutis and the submucosa - the latter can produce obstruction in the upper airways and thereby lead to life-threatening asphyxia. This is the reason for the high, 30-to 50-per-cent mortality of undiagnosed or improperly managed cases. Airway obstruction can be prevented through early diagnosis, meaningful patient information, timely recognition of initial symptoms, state-of-the-art emergency therapy, and close monitoring of the patient. Prophylaxis can substantially mitigate the risk of upper airway edema and also improve the patients' quality of life. Notwithstanding the foregoing, any form of upper airway edema should be regarded as a potentially life-threatening condition. None of the currently available prophylactic modalities is capable of preventing UAE with absolute certainty.


Effects of p-chlorophenylalanine on male sexual behavior in female rats with mesencephalic raphe nuclei lesions.
The role of serotonergic neurons in the mesencephalic raphe nuclei in regulating male sexual behavior in female rats was examined. The median or dorsal raphe nucleus lesions (MRL or DRL) were made in ovariectomized rats and behavioral tests were performed after implantation of Silastic tubes containing testosterone and treatment with serotonin synthesis inhibitor p-chlorophenylalanine (pCPA). Half of the animals in each group received 4 100 mg/kg pCPA injections before the behavioral test. As a result, the incidences and frequencies of mounts and intromissive patterns in the MRL and DRL groups were comparable to those in control females without brain surgery. Mount latency in the MRL females was shorter than that in the control females. When pCPA was given, most females with or without brain surgery showed mounts and intromissive patterns, and frequencies were higher than those in females without pCPA. These results suggest that the median raphe nucleus plays an inhibitory role in the onset mechanism for mounting. On the other hand, a stronger inhibitory influence in regulating male sexual behavior exists in other serotonergic neurons than those in the median and dorsal raphe nuclei in female rats.


Nicotinic pharmacology of anatoxin analogs. I. Side chain structure-activity relationships at peripheral agonist and noncompetitive antagonist sites.
Anatoxin analogs were designed to evaluate the importance of H-bonding, planarity, size and steric configuration of the anatoxin side chain moiety with regard to nicotinic potency and efficacy. This report examines the actions of these analogs on the somatic nicotinic acetylcholine receptor at two different loci: the agonist recognition site and the ion channel site. Agonist effects were evaluated using stimulation of contracture and radioligand binding competition for [125I]alpha bungarotoxin sites in Rana pipiens muscle, and stimulation of [3H]perhydrohistrionicotoxin binding and competition for [125I]alpha bungarotoxin sites in Torpedo californica electric organ. Antagonist effects were evident in the inhibition of neurally evoked twitch of the frog sciatic nerve-sartorius muscle preparation and in inhibition of [3H]perhydrohistrionicotoxin binding to Torpedo receptors. The affinity of these analogs for the agonist locus was consistently associated with activation of the AChR. Our results show that side chain steric configuration has an important role in affinity of the (+)-anatoxin-a analogs for the nicotinic acetylcholine receptor ion channel sites. Several analogs also revealed stereospecific noncompetitive actions. The (+)-anatoxin-a-related structures are important probes for characterizing both agonist and ion channel target sites on the peripheral nicotinic receptor.


Randomised controlled feasibility study of a school-based multi-level intervention to increase physical activity and decrease sedentary behaviour among vocational school students.
No school-based physical activity (PA) interventions among older adolescents have demonstrated long-term effectiveness, and few of them so far have addressed sedentary behaviour (SB). Based on behavioural theories and evidence, we designed a multi-level intervention to increase PA and decrease SB among vocational school students. This study investigates feasibility and acceptability of two main intervention components and research procedures. We also examine uptake of behaviour change techniques (BCTs) by the participants. Design was an outcome assessor blinded, cluster-randomised controlled trial. Four classes of students (matched pairs) were randomised into one intervention and one control arm. The intervention consisted of (1) a 6-h group-based intervention for students, (2) two 2-h training workshops to reduce their students' sitting in class for teachers, and (3) provision of light PA equipment in classrooms. At baseline (T1), mid-intervention (T2) at 3 weeks, post-intervention (T3) and 6 months after baseline (T4) we measured hypothesised psychosocial mediators and self-reported PA and sitting. Objective assessment of PA and SB (7-day accelerometry) was conducted at T1, T3 and T4. Body composition (bioimpedance) was measured at T1 and T4. Students and teachers in the intervention arm filled in acceptability questionnaires at T3. Recruitment rate was 64% (students) and 88.9% (teachers), and at T3, all post-intervention measurements were completed by 33 students (retention 76.7%) and 15 teachers (retention 93.8%). Acceptability ratings of sessions were high (students M = 6.29, scale 1-7), and data collection procedures were feasible. Intervention arm students reported increased use of BCTs, but uptake of some key BCTs was suboptimal. BCT use correlated highly with objective measures of PA. Based on both self-report and student evaluation, teachers in the intervention arm increased the use of sitting reduction strategies at post-intervention and T4 follow-up (p < .05). We detected willingness of the target groups to participate, good response rates to questionnaires, adequate retention, as well as acceptability of the trial protocol. Investigation of BCT use among students helped further enhance intervention procedures to promote BCT use. After making necessary modifications identified, intervention effectiveness can next be tested in a definitive trial. ISRCTN34534846 . Registered 23 May 2014. Retrospectively registered.


In vivo evidence that transcription and splicing are coordinated by a recruiting mechanism.
We describe the nuclear organization of pre-mRNA processing components in HeLa cells upon adenovirus 2 infection and their relationship to the localization of viral RNA sequences. We observe a redistribution of cellular splicing factors as well as RNA polymerase II and heterogeneous nuclear ribonucleoprotein particle proteins to sites of viral RNA transcription. Similar results were obtained in cells transiently transfected with a plasmid containing a portion of the beta-tropomyosin gene. Our findings demonstrate a very close association between RNA transcripts and transcription and pre-mRNA splicing factors, suggesting that these processes are both temporally and spatially linked in the cell nucleus. Furthermore, these data suggest a recruiting mechanism that regulates the localization of transcription and splicing factors in response to the initiation of active transcription.


Anticancer activity of a sub-fraction of dichloromethane extract of Strobilanthes crispus on human breast and prostate cancer cells in vitro.
The leaves of Strobilanthes crispus (S. crispus) which is native to the regions of Madagascar to the Malay Archipelago, are used in folk medicine for their antidiabetic, diuretic, anticancer and blood pressure lowering properties. Crude extracts of this plant have been found to be cytotoxic to human cancer cell lines and protective against chemically-induced hepatocarcinogenesis in rats. In this study, the cytotoxicity of various sub-fractions of dichloromethane extract isolated from the leaves of S. crispus was determined and the anticancer activity of one of the bioactive sub-fractions, SC/D-F9, was further analysed in breast and prostate cancer cell lines. The dichloromethane extract of S. crispus was chromatographed on silica gel by flash column chromatography. The ability of the various sub-fractions obtained to induce cell death of MCF-7, MDA-MB-231, PC-3 and DU-145 cell lines was determined using the LDH assay. The dose-response effect and the EC50 values of the active sub-fraction, SC/D-F9, were determined. Apoptosis was detected using Annexin V antibody and propidium iodide staining and analysed by fluorescence microscopy and flow cytometry, while caspase 3/7 activity was detected using FLICA caspase inhibitor and analysed by fluorescence microscopy. Selected sub-fractions of the dichloromethane extract induced death of MCF-7, MDA-MB-231, PC-3 and DU-145 cells. The sub-fraction SC/D-F9, consistently killed breast and prostate cancer cell lines with low EC50 values but is non-cytotoxic to the normal breast epithelial cell line, MCF-10A. SC/D-F9 displayed relatively higher cytotoxicity compared to tamoxifen, paclitaxel, docetaxel and doxorubicin. Cell death induced by SC/D-F9 occurred via apoptosis with the involvement of caspase 3 and/or 7. A dichloromethane sub-fraction of S. crispus displayed potent anticancer activities in vitro that can be further exploited for the development of a potential therapeutic anticancer agent.


Large-image-format computed tomography imaging spectrometer for fluorescence microscopy.
Multispectral imaging has significantly enhanced the analysis of fixed specimens in pathology and cytogenetics. However, application of this technology to in vivo studies has been limited. This is due in part to the increased temporal resolution required to analyze changes in cellular function. Here we present a non-scanning instrument that simultaneously acquires full spectral information (460 nm to 740 nm) from every pixel within its 2-D field of view (200 ìm x 200 ìm) during a single integration time (typically, 2 seconds). The current spatial and spectral sampling intervals of the spectrometer are 0.985 ìm and 5 nm, respectively. These properties allow for the analysis of physiological responses within living biological specimens.


Social, policy, and public health perspectives on new psychoactive substances.
New psychoactive substances pose a particular challenge to those formulating drugs policy and related public health responses. This paper outlines some of the main issues arising from their use, with a particular focus on user perspectives. Such substances are often (at least initially) produced and distributed for different reasons than controlled drugs. They emerge in users' repertoires undetected by most monitoring systems and general population drug surveys. While reasons for use by innovators and early adopters are often in the spirit of self-experimentation, such substances may rapidly diffuse to the recreational arena as a result of enthusiastic user propagation where they act as substitutes or complements to controlled drugs. The majority of substances are believed to be sourced, albeit not exclusively, from manufacturers based in China. They are retailed to consumers through the Internet and physical shops (such as 'head' and 'smart' shops), as well as traditional 'street dealers' (although data on the significance of this latter route of supply are limited). The data required for risk assessment of the harms such substances may pose, as well as information required for accurate user-derived harm reduction advice, are often limited. Moreover, some involved in the commercial supply have deliberately misbranded products, including substituting the active substance, in apparent attempts to circumvent regulatory frameworks. This leaves users susceptible to both health and criminal justice harms. Despite various attempts to restrict the supply, they often continue to be available through the illicit market, although it is not yet possible to predict whether they will join other drugs such as MDMA and LSD as mainstays of the recreational pharmacopeia.


Trials and intensity effects in single-trial ERP components and autonomic responses in a dishabituation paradigm with very long ISIs.
The phasic orienting reflex (OR) was investigated using single-trial data collected concurrently from 4 autonomic measures and event-related potentials (ERPs). In an auditory dishabituation paradigm, twelve indifferent tones of two intensities (60 or 80 dB, intensity change on trial 11, counterbalanced between subjects) were presented at very long interstimulus intervals (ISIs). Novelty and intensity based stimulus-response patterns were examined seeking ERP analogues of autonomic measures representing pre-OR and OR processing. Skin conductance response (SCR) represented the phasic OR index. EOG-corrected ERP data for 16 subjects were decomposed by a temporal Principal Components Analysis (PCA). SCR diminished over 10 standard trials, recovered on change trial 11, dishabituated to the re-presentation of the standard tone on trial 12, and showed intensity effects at the change - formal requirements for an OR index. The evoked cardiac response (HR) showed no trial or intensity effects. Respiratory pause (RP) decreased linearly over trials and showed recovery but no dishabituation or intensity effect. Peripheral vasoconstriction (PVC) failed to decrement but exhibited an intensity effect. Ten identifiable ERP components were extracted: Na, P1, N1-1, PN, P2, P3a, P3b, a novelty-sensitive HabP3, an intensity-sensitive IntP3, and the Slow Wave (SW). Pattern 1 showed no trial or intensity effects (HR, P1, PN, P2); Pattern 2 showed no trial effect but an intensity effect (PVC, Na, N1-1, P3a); and Pattern 3 demonstrated habituation and an intensity effect (SCR, RP, P3b, HabP3, IntP3, SW). The observed fractionation of autonomic and central measures is consistent with Preliminary Process Theory (PPT) rather than the notion of a unitary OR.


Biofilm facilitates metal accumulation onto microplastics in estuarine waters.
In aquatic environments, plastic debris accumulates chemical pollutants from the surrounding water, potentially altering the fate of xenobiotics in these ecosystems. The effects of biofouling on the potential for plastic to sorb environmental pollutants remain poorly understood. In this study, we test the hypothesis that concentrations of metals are directly related to biofilm accumulation on microplastics submerged in natural estuarine waters. Two types of pre-production plastic pellets (polylactic acid (PLA) and low-density polyethylene (LDPE)) and glass pellets, were suspended for up to 28 days in an urbanized estuary (San Francisco Bay, California) to investigate how biofilm affects the accumulation of metals on these materials. During the initial weeks of the experiment, biofilm growth differed between locations, but after 28 days, PLA and LDPE had similar amounts of biofilm at the two field sites. Biofilm was the only significant predictor variable for Ba, Cs, Fe, Ga, Ni and Rb, and simple regressions of these metals after one month of submersion predicted much of the variability in the data (respective adjusted R2 values: 0.46, 0.90, 0.86, 0.81, 0.87, 0.90; p < 0.001). For other metals influenced by location or substrate material, multivariate analysis showed that increases in metal concentrations were predicted by increases in biofilm for Cu, Pb, Al, K, U, Co, Mg (p < 0.001) and Mn (p < 0.01). This work highlights the role of biofilm in facilitating metal accumulation on plastic debris and contributes to current understanding of the underlying processes that influence the behavior of microplastics as aquatic contaminants.


Hepatitis C virus core protein interacts with cellular putative RNA helicase.
The nucleocapsid core protein of hepatitis C virus (HCV) has been shown to trans-act on several viral or cellular promoters. To get insight into the trans-action mechanism of HCV core protein, a yeast two-hybrid cloning system was used for identification of core protein-interacting cellular protein. One such cDNA clone encoding the DEAD box family of putative RNA helicase was obtained. This cellular putative RNA helicase, designated CAP-Rf, exhibits more than 95% amino acid sequence identity to other known RNA helicases including human DBX and DBY, mouse mDEAD3, and PL10, a family of proteins generally involved in translation, splicing, development, or cell growth. In vitro binding or in vivo coimmunoprecipitation studies demonstrated the direct interaction of the full-length/matured form and C-terminally truncated variants of HCV core protein with this targeted protein. Additionally, the protein's interaction domains were delineated at the N-terminal 40-amino-acid segment of the HCV core protein and the C-terminal tail of CAP-Rf, which encompassed its RNA-binding and ATP hydrolysis domains. Immunoblotting or indirect immunofluorescence analysis revealed that the endogenous CAP-Rf was mainly localized in the nucleus and to a lesser extent in the cytoplasm, and when fused with FLAG tag, it colocalized with the HCV core protein either in the cytoplasm or in the nucleus. Similar to other RNA helicases, this cellular RNA helicase has nucleoside triphosphatase-deoxynucleoside triphosphatase activity, but this activity is inhibited by various forms of homopolynucleotides and enhanced by the HCV core protein. Moreover, transient expression of HCV core protein in human hepatoma HuH-7 cells significantly potentiated the trans-activation effect of FLAG-tagged CAP-Rf or untagged CAP-Rf on the luciferase reporter plasmid activity. All together, our results indicate that CAP-Rf is involved in regulation of gene expression and that HCV core protein promotes the trans-activation ability of CAP-Rf, likely via the complex formation and the modulation of the ATPase-dATPase activity of CAP-Rf. These findings provide evidence that HCV may have evolved a distinct mechanism in alteration of host cellular gene expression regulation via the interaction of its nucleocapsid core protein and cellular putative RNA helicase known to participate in all aspects of cellular processes involving RNA metabolism. This feature of core protein may impart pleiotropic effects on host cells, which may partially account for its role in HCV pathogenesis.


Are history and physical examination a good screening test for sleep apnea?
To determine whether presenting clinical history, pharyngeal examination, and the overall subjective impression of the clinician could serve as a sensitive screening test for sleep apnea. Blinded comparison of history and physical examination with results of nocturnal polysomnography. Sleep clinic of a tertiary referral center. A total of 410 patients referred for suspected sleep apnea syndrome. Most patients reported snoring. All patients were asked standard questions and given an examination relevant to the diagnosis of the sleep apnea syndrome, and all had full nocturnal polysomnography. Patients with more than ten episodes of apnea or hypopnea per hour of sleep were classified as having sleep apnea. Stepwise linear logistic regression was used to develop two predictive models of sleep apnea: one based on the presence of characteristic clinical features, age, sex, and body mass index; and one based on subjective clinical impression. The prevalence of sleep apnea in our patients was 46%. Only age, body mass index, male sex, and snoring were found to be predictors of sleep apnea. The logistic rule discriminated between patients with and without sleep apnea (receiver operating characteristic [ROC] area, 0.77 [95% Cl, 0.73 to 0.82]). For patients with a predicted probability of apnea of less than 20%, the clinical model had 94% sensitivity and 28% specificity. Subjective impression alone identified correctly only 52% of patients with sleep apnea and had a specificity of 70%. In patients with a high predicted probability of the sleep apnea syndrome, subjective impression alone or any combination of clinical features cannot serve as a reliable screening test. However, in patients with a low predicted probability of sleep apnea, the model based on clinical data was sufficiently sensitive to permit about a 30% reduction in the number of unnecessary sleep studies.


Differential parsing of EGFR endocytic flux among parallel internalization pathways in lung cancer cells with EGFR-activating mutations.
Due to the existence of parallel pathways for receptor endocytosis and their complexities, a quantitative understanding of receptor endocytosis in normal and pathological settings requires computational analysis. Here, we develop a mechanistic model of epidermal growth factor receptor (EGFR) endocytosis to determine the relative contributions of three parallel pathways: clathrin-dependent internalization mediated by mitogen-inducible gene 6 (MIG6), an endogenous EGFR kinase inhibitor that links EGFR to endocytic proteins; clathrin-dependent internalization mediated by the ubiquitin ligase CBL, which can be sequestered by the regulatory protein Sprouty2; or alternative pathways that may be non-clathrin mediated. We applied the model to interpret our previous measurements of EGFR endocytosis in lung cancer cells. Interestingly, our results suggest that MIG6 is responsible for at least as much wild-type EGFR internalization as CBL, indicating that a significant fraction of internalizing EGFR may be incapable of driving signaling. Model results also suggest that MIG6's endocytic function is reduced for the kinase-activated and internalization-impaired EGFR mutants found in some lung cancers. Analysis of Sprouty2 knockdown data indicates that Sprouty2 regulates EGFR endocytosis primarily by controlling EGFR expression, rather than by sequestering CBL, and supports the notion that CBL-mediated internalization is impaired for EGFR mutants. We further demonstrate that differences in internalization between wild-type and mutant EGFR cannot explain differences in EGF-mediated EGFR degradation without concomitant changes in EGFR recycling, which we previously quantified. This work provides new quantitative insights into EGFR trafficking in lung cancer and provides a framework for studying parallel endocytosis pathways for other receptors.


Restoring multidrug resistance-associated protein 3 attenuates cell proliferation in the polycystic kidney.
Autosomal dominant polycystic kidney disease (ADPKD) is characterized by abnormal proliferation of renal tubular epithelial cells, resulting in the loss of renal function. Despite identification of the genes responsible for ADPKD, few effective drugs are currently available for the disease. Thus finding additional effective drug targets is necessary. The functions of multidrug- resistance-associated protein 3 (MRP3) have been reported only in the field of drug resistance, and the renal functions of MRP3 are mostly unknown. In this study, we found that MRP3 was significantly downregulated in kidneys of human patients with ADPKD and polycystic kidney disease (PKD) mouse models. Our results suggest that downregulated MRP3 stimulated renal epithelial cell proliferation through the B-Raf/MEK/ERK signaling pathway. In contrast, we found that restoring MRP3 reduced cell proliferation and cystogenesis in vitro. These results suggest that the renal function of MRP3 is related to renal cell proliferation and cyst formation and that restoring MRP3 may be an effective therapeutic approach for PKD.


A Phase II Study of Alisertib in Children with Recurrent/Refractory Solid Tumors or Leukemia: Children's Oncology Group Phase I and Pilot Consortium (ADVL0921).
Aurora A kinase (AAK) plays an integral role in mitotic entry, DNA damage checkpoint recovery, and centrosome and spindle maturation. Alisertib (MLN8237) is a potent and selective AAK inhibitor. In pediatric preclinical models, antitumor activity was observed in neuroblastoma, acute lymphoblastic leukemia, and sarcoma xenografts. We conducted a phase 2 trial of alisertib in pediatric patients with refractory or recurrent solid tumors or acute leukemias (NCT01154816). Alisertib (80 mg/m2/dose) was administered orally, daily for 7 days every 21 days. Pharmacogenomic (PG) evaluation for polymorphisms in the AURK gene and drug metabolizing enzymes (UGT1A1*28), and plasma pharmacokinetic studies (PK) were performed. Using a 2-stage design, patients were enrolled to 12 disease strata (10 solid tumor and 2 acute leukemia). Response was assessed after cycle 1, then every other cycle. A total of 139 children and adolescents (median age, 10 years) were enrolled, 137 were evaluable for response. Five objective responses were observed (2 complete responses and 3 partial responses). The most frequent toxicity was myelosuppression. The median alisertib trough concentration on day 4 was 1.3 μmol/L, exceeding the 1 μmol/L target trough concentration in 67% of patients. No correlations between PG or PK and toxicity were observed. Despite alisertib activity in pediatric xenograft models and cogent pharmacokinetic-pharmacodynamic relationships in preclinical models and adults, the objective response rate in children and adolescents receiving single-agent alisertib was less than 5%.


Long-term transgene expression and inhibition of HIV-1 replication by a Cre/loxP-EBNA-1/oriP HIV-1-dependent ribozyme vector: Applications for HIV-1 gene therapy.
The cleavage of target mRNA by ribozymes is being exploited as a means of gene silencing in nucleic-acid-based therapies. We previously established an HIV-1-dependent ribozyme-expression vector system, based on Cre-loxP technology with an LTR-gag-p17 promoter as a molecular switch for use in acute HIV-1 infection. The simultaneous expression of the Cre protein and loxP homologous recombination induced a high level of HIV-1-replication inhibition, but ribozyme expression was transient. In the current study, we overcame this limitation by inserting EBNA-1 and oriP genes from the Epstein-Barr virus (EBV) into the vector. When this plasmid was introduced into HeLa CD4(+) cells, we observed long-term expression of both the EGFP reporter gene and the ribozyme. Moreover, HIV-1 replication was inhibited in the long-term in transfected cells. These data suggest that the HIV-1-dependent ribozyme-expression vector containing EBNA-1/oriP sequences would be a useful tool in HIV-1 gene therapy applications.


TRPC3 channels are required for synaptic transmission and motor coordination.
In the mammalian central nervous system, slow synaptic excitation involves the activation of metabotropic glutamate receptors (mGluRs). It has been proposed that C1-type transient receptor potential (TRPC1) channels underlie this synaptic excitation, but our analysis of TRPC1-deficient mice does not support this hypothesis. Here, we show unambiguously that it is TRPC3 that is needed for mGluR-dependent synaptic signaling in mouse cerebellar Purkinje cells. TRPC3 is the most abundantly expressed TRPC subunit in Purkinje cells. In mutant mice lacking TRPC3, both slow synaptic potentials and mGluR-mediated inward currents are completely absent, while the synaptically mediated Ca2+ release signals from intracellular stores are unchanged. Importantly, TRPC3 knockout mice exhibit an impaired walking behavior. Taken together, our results establish TRPC3 as a new type of postsynaptic channel that mediates mGluR-dependent synaptic transmission in cerebellar Purkinje cells and is crucial for motor coordination.


Associations of tibial lead levels with BsmI polymorphisms in the vitamin D receptor in former organolead manufacturing workers.
We evaluated associations of tibial lead levels with polymorphisms in the vitamin D receptor (VDR) in 504 former organolead manufacturing workers with past exposure to lead. In this cross-sectional study, we measured tibial lead by (109)Cd K-shell X-ray fluorescence. Tibial lead was evaluated in subjects with different VDR genotypes defined using the BsmI restriction enzyme, adjusting for confounding variables. Study participants had a mean age +/- SD of 57.4 +/- 7.6 years. A total of 169 (33.5%) subjects were homozygous for the BsmI restriction site (designated bb), 251 (49.8%) were heterozygous (Bb), and 84 (16.7%) were homozygous for the absence of the restriction site (BB). Among all of the study subjects, tibial lead concentrations were low, with a mean +/- SD of 14.4 +/- 9.3 microg Pb/g bone mineral. There were only small differences in tibial lead concentrations by VDR genotype, with mean +/- SD tibial lead concentrations of 13.9 +/- 7.9, 14.3 +/- 9.5, and 15.5 +/- 11.1 in subjects with bb, Bb, and BB, respectively. In a multiple linear regression model of tibial lead concentrations, the VDR genotype modified the relation between age and tibial lead concentrations; subjects with the B allele had larger increases in tibial lead concentrations with increasing age (0.37, 0.48, and 0.67 microg/g per year of age in subjects with bb, Bb, and BB, respectively; the adjusted p-value for trend in slopes = 0.04). The VDR genotype also modified the relation between years since last exposure to lead and tibial lead concentrations. Subjects with bb evidenced an average decline in tibial lead concentrations of 0.10 microg/g per year since their last exposure to lead, whereas subjects with Bb and BB evidenced average increases of 0.03 and 0.11 microg/g per year, respectively (the adjusted p-value for trend in slopes = 0.01). Polymorphisms in the vitamin D receptor modified the relations of age and years since the last exposure to lead with tibial lead concentrations. Although controversy remains on the influence of the VDR genotype on bone mineral density, the data suggest that variant VDR alleles modify lead concentrations in bone, either by influencing lead content or calcium content or both.


Protective immunity against infection with Mycoplasma haemofelis.
Hemoplasmas are potentially zoonotic mycoplasmal pathogens, which are not consistently cleared by antibiotic therapy. Mycoplasma haemofelis is the most pathogenic feline hemoplasma species. The aim of this study was to determine how cats previously infected with M. haemofelis that had recovered reacted when rechallenged with M. haemofelis and to characterize the immune response following de novo M. haemofelis infection and rechallenge. Five specific-pathogen-free (SPF)-derived naive cats (group A) and five cats that had recovered from M. haemofelis infection (group B) were inoculated subcutaneously with M. haemofelis. Blood M. haemofelis loads were measured by quantitative PCR (qPCR), antibody response to heat shock protein 70 (DnaK) by enzyme-linked immunosorbent assay (ELISA), blood lymphocyte cell subtypes by flow cytometry, and cytokine mRNA levels by quantitative reverse transcriptase PCR. Group A cats all became infected with high bacterial loads and seroconverted, while group B cats were protected from reinfection, thus providing the unique opportunity to study the immunological parameters associated with this protective immune response against M. haemofelis. First, a strong humoral response to DnaK was only observed in group A, demonstrating that an antibody response to DnaK is not important for protective immunity. Second, proinflammatory cytokine interleukin-6 (IL-6) mRNA levels appeared to increase rapidly postinoculation in group B, indicating a possible role in protective immunity. Third, an increase in IL-12p35 and -p40 mRNA and decrease in the Th2/Th1 ratio observed in group A suggest that a Th1-type response is important in primary infection. This is the first study to demonstrate protective immunity against M. haemofelis reinfection, and it provides important information for potential future hemoplasma vaccine design.


Noninvasive inference of the molecular chemotactic response using bacterial trajectories.
The quality of sensing and response to external stimuli constitutes a basic element in the selective performance of living organisms. Here we consider the response of Escherichia coli to chemical stimuli. For moderate amplitudes, the bacterial response to generic profiles of sensed chemicals is reconstructed from its response function to an impulse, which then controls the efficiency of bacterial motility. We introduce a method for measuring the impulse response function based on coupling microfluidic experiments and inference methods: The response function is inferred using Bayesian methods from the observed trajectories of bacteria swimming in microfluidically controlled chemical fields. The notable advantages are that the method is based on the bacterial swimming response, it is noninvasive, without any genetic and/or mechanical preparation, and assays the behavior of the whole flagella bundle. We exploit the inference method to measure responses to aspartate and α-methylaspartate--measured previously by other methods--as well as glucose, leucine, and serine. The response to the attractant glucose is shown to be biphasic and perfectly adapted, as for aspartate. The response to the attractant serine is shown to be biphasic yet imperfectly adapted, that is, the response function has a nonzero (positive) integral. The adaptation of the response to the repellent leucine is also imperfect, with the sign of the two phases inverted with respect to serine. The diversity in the bacterial population of the response function and its dependency upon the background concentration are quantified.


Optical studies of the secretory event at vertebrate nerve terminals.
Potentiometric probes are small (300-500 Mr) amphipatic molecules that bind to, but do not cross, cell membranes and behave as fast linear transducers of membrane voltage. Their optical properties, particularly absorbance and fluorescence, respond to changes in potential in less than 2 microseconds, and they may be used to follow electrical events in membranes which are inaccessible to microelectrodes. We have used these dyes to study the properties of the action potential in the neurosecretory terminals of vertebrate neurohypophyses and, in particular, to investigate the behaviour of the local population of calcium channels. These channels are sensitive to the peptide toxin omega-conotoxin GVIA, derived from the venom of the marine snail Conus geographicus, but insensitive to dihydropyridine channel modulators. In the neurohypophysis of the mouse, it is possible to demonstrate that the calcium channels that are blocked by omega-conotoxin are those that are required for secretion of peptide hormones. In the terminals of the neurohypophysis, excitation is coupled to secretion, and the secretory event is accompanied by large and rapid changes in light scattering. These intrinsic optical signals provide a millisecond time-resolved monitor of events in the terminal that follow the entry of calcium, and may precede the release of hormones. We will consider how the changes in light scattering can be related to secretion, and how the extrinsic (absorption) and intrinsic optical signals may provide complementary information about excitation-secretion coupling.


Intermolecular protein interactions in solutions of calf lens alpha-crystallin. Results from 1/T1 nuclear magnetic relaxation dispersion profiles.
From analyses of the magnetic field dependence of 1/T1 (NMRD profiles) of water protons in solutions of calf lens alpha-crystallin at several concentrations, we find two regimes of solute behavior in both cortical and nuclear preparations. Below approximately 15% vol/vol protein concentration, the solute molecules appear as compact globular proteins of approximately 1,350 (cortical) and approximately 1,700 (nuclear) kD. At higher concentrations, the effective solute particle size increases, reversibly, as evidenced by the appearance of spectra-like 14N peaks in the NMRD profiles and a change in the field and temperature dependence of 1/T1. At these higher concentrations, the profiles are very similar to those of calf gamma II-crystallin, a crystallin that undergoes an analogous transition near approximately 15% protein (Koenig, S. H., C.F. Beaulieu, R. D. Brown III, and M. Spiller, 1990. Biophys. J. 57:461-469). By comparison with recent analyses of NMRD results for solutions of immobilized proteins as models for the transition from protein solutions to tissue (Koenig, S. H., and R. D. Brown III. 1991. Prog. NMR Spectr. 22:487-567), we argue that alpha-crystallin solute behaves as aggregates approximately greater than 50,000 kD as protein concentration is progressively increased above 15%. Finally, the concentration dependence of the NMRD profiles of alpha- and gamma II-crystallin can readily explain recent osmotic pressure data, in particular the intersection of the respective pressure curves at approximately 23% vol/vol (Vérétout, F., and A. Tardieu. 1989. Eur. Biophys. J. 17:61-68).


A 57,000-mol-wt protein uniquely present in nonproliferating cells and senescent human fibroblasts.
Mouse monoclonal antibody, S-30, was produced from hybridoma preparation from mice injected with the cytoskeleton extract of an in vitro aged culture of human fibroblasts derived from a 66-yr-old donor. The antibody stains positively the nuclei of the nonproliferating cells present predominantly in the senescent cultures of five selected fibroblast strains derived from donors of different age groups, whereas a negative reaction is observed in the cultures of their young counterparts. In the intermediate stage of the in vitro life span of these cell strains, a heterogeneous positive reaction for staining with S-30 antibody is observed in different subfractions of cell cultures. However, the expression of S-30 can be induced in the young fibroblasts at the early stage of their life by prolonged culturing to confluence. This induced expression of S-30 nuclear staining can be depleted upon subculturing at low cell density. Immunoelectron microscopy with colloidal gold-protein A complex demonstrates that the S-30 proteins are present in the nuclear plasma and at the region of nuclear envelope in a clustered arrangement. Immunoprecipitation of [3H]leucine labeled cell specimens shows that the antibody S-30 reacts with a protein with a molecular weight of approximately 57,000.


FoxO1-mediated autophagy plays an important role in the neuroprotective effects of hydrogen in a rat model of vascular dementia.
Vascular dementia (VD) is a heterogeneous group of brain disorders in which cognitive impairment is attributed to cerebrovascular pathologies. Autophagy, a self-cannibalization mechanism, has been demonstrated to be involved in VD progression. Molecular hydrogen is known for its powerful anti-oxidative, anti-apoptotic, and anti-inflammatory activities, and it is also involved in autophagy. However, the effects of hydrogen on VD remain unclear. The current study found that hydrogen-rich water (HRW) significantly alleviated spatial learning and memory impairments. Similar to donepezil treatment, HRW also inhibited neuron loss and shrinkage in the hippocampal CA1 region. In addition, we found that HRW significantly increased the Bcl-2/Bax expression ratio and decreased cleaved caspase-3 expression levels in the hippocampus of VD rats. Moreover, electron microscopy revealed that HRW decreased the number of autophagosomes. We also observed that HRW reduced the increased ratio of LC3-II/I and Beclin 1 expression and saliently upregulated p62 expression. Furthermore, FoxO1 (a major mediator of autophagy regulation) and Atg7 levels were apparently decreased in the hippocampus of HRW-treated bilateral common carotid artery occlusion (2VO) rats. Taken together, these data show that molecular hydrogen exerts beneficial effects on cognitive impairment induced by chronic cerebral hypoperfusion. FoxO1-mediated autophagy plays an important role in the neuroprotective effects of hydrogen in a rat model of VD. Furthermore, the present findings highlight that HRW should be further investigated as a new therapeutic strategy for VD treatment in the future.


A one-pot multicomponent coupling/cyclization for natural product herbicide (±)-thaxtomin A.
Herbicide (±)-thaxtomin A has been synthesized in a one-pot process with a 32% isolated yield. A multicomponent coupling reaction was utilized to prepare in situ a dipeptide precursor which then sequentially underwent an alkaline mediated keto-amide cyclization to provide the target molecule. Adjustment of diastereoselectivity was achieved using microwave-induced irradiation. The approach incorporates atom economy and reaction efficiency and allows for facile library development.


Estimating the support of a high-dimensional distribution.
Suppose you are given some data set drawn from an underlying probability distribution P and you want to estimate a "simple" subset S of input space such that the probability that a test point drawn from P lies outside of S equals some a priori specified value between 0 and 1. We propose a method to approach this problem by trying to estimate a function f that is positive on S and negative on the complement. The functional form of f is given by a kernel expansion in terms of a potentially small subset of the training data; it is regularized by controlling the length of the weight vector in an associated feature space. The expansion coefficients are found by solving a quadratic programming problem, which we do by carrying out sequential optimization over pairs of input patterns. We also provide a theoretical analysis of the statistical performance of our algorithm. The algorithm is a natural extension of the support vector algorithm to the case of unlabeled data.


New chemistry of 1,2-closo-P2B10H10 and 1,2-closo-As2B10H10; in silico and gas electron diffraction structures, and new metalladiphospha- and metalladiarsaboranes.
The molecular structures of 1,2-closo-P(2)B(10)H(10) (1) and 1,2-closo-As(2)B(10)H(10) (2) have been determined by gas electron diffraction and the results obtained compared with those from computation at the MP2/6-31G** level of theory. The level of agreement is good for 2 (root-mean-square [rms] misfit for As and B atoms 0.0297 Å) and very good for 1 (rms misfit for P and B atoms 0.0082 Å). In comparing the structures of 1 and 2 with that of 1,2-closo-C(2)B(10)H(12) (I) it is evident that expansion of the polyhedron from I to 1 to 2 is restricted only to the heteroatom vertices and the B(6) face to which these are bound. Following deboronation (at B3) and subsequent metallation, compounds 1 and 2 have been converted into the new metalladiheteroboranes 3-(η-C(9)H(7))-3,1,2-closo-CoAs(2)B(9)H(9) (4), 3-(η-C(10)H(14))-3,1,2-closo-RuAs(2)B(9)H(9) (5), 3-(η-C(5)H(5))-3,1,2-closo-CoP(2)B(9)H(9) (6), 3-(η-C(9)H(7))-3,1,2-closo-CoP(2)B(9)H(9) (7) and 3-(η-C(10)H(14))-3,1,2-closo-RuP(2)B(9)H(9) (8), the last three constituting the first examples of metalladiphosphaboranes. Together with the known compound 3-(η-C(5)H(5))-3,1,2-closo-CoAs(2)B(9)H(9) (3), compounds 4-8 have been analysed by NMR spectroscopy and (except for 8) single-crystal X-ray diffraction. The (11)B NMR spectra of analogous pairs of metalladiphosphaborane and metalladiarsaborane (6 and 3, 7 and 4, 8 and 5) reveal a consistently narrower (9-10 ppm) chemical shift range for the metalladiarsaboranes, the combined result of a deshielding of the lowest frequency resonance (B6) and an increased shielding of the highest frequency resonance (B8) via an antipodal effect. In crystallographic studies, compounds 3 and 5B (one of two crystallographically-independent molecules) suffer As/B disorder, but in both cases it was possible to refine distinct, ordered, components of the disorder, the first time this has been reported for metalladiarsaboranes. Moreover, whilst the Cp compounds 6 and 3 are disordered, their indenyl analogues 7 and 4 are either ordered or significantly less disordered, a consequence of both the reduced symmetry of an indenyl ligand compared to a Cp ligand and the preference of the former for a distinct conformation relative to the cage heteroatoms. Unexpectedly, whilst this conformation in the cobaltadiphosphaborane 7 is cis-staggered (similar to that previously established for the analogous cobaltadicarborane), in the cobaltadiarsaborane 4 the conformation is close to cis-eclipsed.


Class I alcohol dehydrogenase is highly expressed in normal human mammary epithelium but not in invasive breast cancer: implications for breast carcinogenesis.
Detoxification of ethanol can contribute to oxidative cellular and DNA damage and, thereby, to carcinogenesis. The potential relevance of this to breast carcinogenesis is suggested by evidence that alcohol consumption is a risk factor for breast cancer. It is, however, not known whether ethanol can be metabolized in breast parenchyma. The goal of this study was to determine whether class I and/or IV alcohol dehydrogenase (ADH), medium chain ADHs that can catalyze oxidation of ethanol, are expressed in human breast parenchyma. Normal and neoplastic human breast tissue specimens were examined for class I and IV ADH mRNA by reverse transcription-PCR, for protein by immunocytochemistry and Western analysis, and for their potential to catalyze NAD(+)-dependent oxidation of ethanol. Together, the findings provide evidence that: (a) class I ADH is the medium-chain ADH that is expressed in human breast parenchyma, specifically in the mammary epithelium; (b) human breast parenchyma can support ADH-mediated oxidation of ethanol; and (c) the expression of class I ADH is dramatically reduced or abrogated in invasive breast cancers. Expression of class I ADH in normal human breast parenchyma was confirmed by probing a multiple human tissue polyA(+)RNA. The unexpected finding of virtual abrogation of expression of class I ADH in invasive breast cancer suggests that the enzyme has some "tumor suppressor" function in the mammary epithelium. The one property of class I ADH fitting this designation is its potential to catalyze the oxidation of the micronutrient/prohormone retinol to retinal, the first step in the biosynthesis of retinoic acid, the principal known mediator of the actions of retinoids important for maintaining epithelia in a differentiated state.


Intimate Partner Violence and Psychological Distress Among Emerging Adult Women: A Bidirectional Relationship.
Background: Intimate partner violence (IPV) and psychological distress (PD) are major public health concerns among emerging adult women. Emerging adulthood presents a complex set of new experiences and challenges that pose a risk to normative development. In particular, an increased prevalence of IPV and PD during this time period may lead to long-term health consequences. Methods: Data from the Relationship Dynamics and Social Life study, a longitudinal study of a racially and socioeconomically diverse population-representative random sample of 726 partnered women, aged 18-19, residing in a Michigan county, and followed for 2.5 years, were used to investigate the relationship between IPV and PD. Logistic regression models predicted each measure of PD (depression, stress, loneliness, self-esteem) as a function of past IPV (none, psychological violence only, any physical violence), and multinomial logistic regression models predicted subsequent weekly IPV as a function of each measure of PD. Results: PD and IPV were prevalent among emerging adult women. Past psychological IPV was associated with experiencing all four distress measures. Past physical IPV was also associated with depression, stress, and loneliness, but not self-esteem. Women with each PD were more likely to subsequently experience psychological violence, and women who reported stress were more likely to subsequently experience any physical violence. Conclusions: The IPV-PD relationship is bidirectional. Women who experienced past IPV were more likely to report PD. Conversely, women who experienced PD were at a greater risk of subsequent IPV.


A nanoemulsion of an anti-oxidant synergy formulation reduces tumor growth rate in neuroblastoma-bearing nude mice.
Neuroblastoma, the most common form of childhood cancer, may arise from a biochemical block of cellular differentiation and a resultant continuation of a proliferative state. Neuroblastoma often spontaneously reverts by undergoing partial differentiation and ultimate degeneration and may be associated with the generation of reactive oxygen species (ROS). We have recently reported in neuroblastoma cell culture studies that an anti-oxidant synergy formulation (ASF) can induce differentiation and buffer neuronal degeneration and oxidative stress in cultured cortical neurons and in central nervous system tissue of apolipoprotein E-deficient mice. The objective of the present study was to investigate whether a subcutaneous injection and/or transdermal application of a nanoemulsion preparation of ASF would reduce tumor growth rate in a neuroblastoma xenograph mouse model. The results indicate that whereas suspensions of ASF were ineffective in decreasing tumor growth rate in the neuroblastoma mouse model, tumor growth rate was similarly reduced an average 65% by either subcutaneous injection or transdermal application of an ASF nanoemulsion preparation to the tumor. In conclusion, the data suggest that subcutaneous and/or transdermal application of an ASF nanoemulsion preparation is effective in reducing tumor growth rate in this neuroblastoma mouse model.


Localization of aggregated cell surface antigens of target cells bound to cytotoxic T lymphocytes.
The redistribution of aggregated cell surface antigens of target cells bound to cytotoxic T lymphocytes was investigated. It was found that cap formation induced by antibody always occurred toward the site of binding. It is suggested that the polar localization of capped target cell surface determinants is a consequence of cytotoxic T-lymphocyte target cell interaction.


Initial evaluation of use of an online partner notification tool for STI, called 'suggest a test': a cross sectional pilot study.
Partner notification is crucial for sexually transmitted infection (STI) control. We developed Suggestatest.nl (SAT), an internet-based notification system for verified diagnoses of STI/HIV. SAT uses email, short message service, postal letter or a gay dating site to notify sexual contacts. SAT was piloted at the Public Health STI clinics in two major cities in the Netherlands. We evaluated SAT from March to July 2012 by analysing SAT notifications linked with epidemiological data. Determinants for SAT use were assessed using multivariable logistic regression analysis. Of 988 index clients receiving a SAT code, overall 139 (14%) notified through SAT, sending 505 notifications (median 2), 84% by text messaging and 15% by email; 88% non-anonymously. Of those intending to use SAT, 23% notified with SAT. Intention to use SAT was the only independent determinant of SAT use in heterosexuals and men who have sex with men. Among the 67 SAT users in Rotterdam, 56% (225/402) of their partners at risk were contactable, and 95% (213/225) of those were notified using SAT. 58% of SAT-notified partners accessed the SAT-website and 20% of them subsequently consulted the STI clinics. STI positivity in partners was lower in those notified by SAT (28% (32/116)) than in those with contact cards (45% (68/152); p<0.001). Although the challenges posed by non-contactable partners are not solved by SAT, it is a valuable novel tool for notification of verified STI diagnoses by index patients and providers. In addition to current standard partner notification practice it suits a small number of clients, especially those reporting more than one partner.


Workplace injuries in the Australian allied health workforce.
Objective This study aims to identify the number, costs and reported injury mechanisms of serious injury claims for allied health professionals. Methods Using Australian Workers' Compensation injury data, the number, mechanism, and costs of injury claims were calculated for eight groups of allied health professions (chiropractors and osteopaths, speech pathologists and audiologists, occupational therapists, physiotherapists, psychologists, podiatrists, social workers and prosthetists/orthotists) between the 2000-01 and 2013-14 financial years. Workforce injury rates were calculated using the 2011 Australian Census Workforce data (denominator) and 2011 Workers' Compensation Statistics claims data (numerator). Results Across the allied health professions, 7023 serious injuries (minimum 5 days absence from work) were recorded with an associated total compensation cost of A$201970000. Fewer than 1.5% of each allied health professional group had an injury claim, with the exception of prosthetists/orthotists who had a rate of 25.9% serious injury claims (95% confidence interval 21.9-30.4). The average cost per claim varied across the allied health professions, from the lowest cost of A$19091 per injury for occupational therapists to the highest of A$48466 per claim in chiropractic and osteopathy. Body stressing followed by mental stress were the most common mechanisms of injury. Conclusions Mechanism of injury, both physical and psychosocial, were identified. Prosthetists/orthotists are at the highest risk of workplace injury of all allied health professions. This suggests the need for further investigation and development of appropriately targeted injury prevention programs for each allied health profession. What is known about this topic? Retention of allied health professionals is a significant issue, with workplace injuries identified as one contributing factor to this problem. Healthcare workers are potentially at high risk of injury as they are exposed to a range of physical and psychosocial hazards in their workplace. What does this paper add? This paper is the first to report on serious injuries, minimum 5 days absence from work, from Australian Workers' Compensation data, across a range of allied health professions. Various allied health professions were examined to identify the number, mechanism and cost of serious workplace injuries finding there is an average of 500 serious claims per year at a cost of A$14million. Prosthetists/orthotists were identified as having the highest proportion of claims per workforce population. What are the implications for practitioners? These results suggest highly varied injury rates across allied health professions. Compensation data does not enable accurate identification of causal factors. Further work is required to identify relevant causal factors so that targeted risk reduction strategies can be developed to reduce workforce injuries.


The clinical conscientiousness index: a valid tool for exploring professionalism in the clinical undergraduate setting.
The need to develop effective tools to measure professionalism continues to challenge medical educators; thus, as a follow-up to a recent examination of the "Conscientiousness Index" (CI, a novel measure of one facet of professionalism) in one setting with preclinical medical students, the authors aimed to investigate the validity of the CI as a proxy measure of professionalism in a different context and in the clinical phase of undergraduate medical education. In academic year 2009-2010, the authors collected data similar to those collected for the original preclinical study. In an effort to create a Clinical Conscientiousness Index (CCI) score, they collected the following information on 124 third-year medical students completing their clinical rotations: attendance, timeliness of assessment submissions, and completion of rotation evaluations. Then, they compared the resultant CCI scores with faculty views on professionalism and with formal assessments of students' professionalism (i.e., their portfolios and objective structured clinical examinations [OSCEs]). The authors demonstrate significant correlations between CCI scores and faculty views on professionalism (rS = 0.3; P = .001), and between CCI scores and OSCE score (rS = 0.237; P = .008), but not between CCI scores and portfolio assessment (rS = 0.084; P = .354). The authors also present relationships between CCI scores and demographics. The CCI is a practical, valid proxy measure of professionalism, achieving good correlation with faculty views on professionalism and clinical competency examinations, but not portfolio assessment, in one clinical undergraduate setting.


In vitro comparison of four different electronic apex locators to determine the major foramen using the clearing technique.
The aim of this study was to evaluate the accuracy of four electronic apex locators (EALs) (DentaPort ZX, Raypex 5, Endo Master and VDW Gold) in detecting the major foramen using the clearing technique. Forty-eight extracted single-rooted extracted teeth with mature apices were used for the study and divided into four groups of 12 teeth each. All teeth were embedded in an alginate model. Electronic measurements were taken using a size 15 K-file attached to the holder. Then, the teeth were cleared and photographed under a stereomicroscope with a digital camera. The distance between the tip of the file and the major foramen was measured by using an image analysis software program. Positive and negative values were recorded when the file tip was beyond or short of the major foramen and zero value when the file tip and the major foramen coincided. Statistical analysis was performed using the Kruskal-Wallis and Chi-square tests at a significance level of 0.05. Mean distance from the file tip to the major foramen were 0.302 ± 0.202, 0.065 ± 0.293, 0.117 ± 0.475, and 0.258 ± 0.160 mm in the DentaPort ZX, Raype 5, Endo Master, and VDW Gold groups, respectively. There were no statistically significant differences among the devices (P > 0.05) . Under the experimental conditions, all EALs showed an acceptable determination of the major foramen.


Rationing health care: it's a matter of the health care system's structure.
The article examines two primary policy proposals for how the U.S. should allocate its limited health care dollars: a centralized model in which a commission establishes rationing guidelines, and a decentralized model in which rationing decisions are made by health care providers on a case by case basis. The author finds significant advantages with each position, leading the author to assert that a combination of each is key to an effective rationing policy: a centralized control of structure coupled with decentralized physician-level decision making. While mindful that formal rationing guidelines alone are unfeasible to effectuate cost-effective care, the author introduces two decentralized policies to control costs: the limitation of resources at physicians' disposal and elimination of physicians' personal incentive to provide high-cost care.


Insights into study design and statistical analyses in translational microbiome studies.
Research questions in translational microbiome studies are substantially more complex than their counterparts in basic science. Robust study designs with appropriate statistical analysis frameworks are pivotal to the success of these translational studies. This review considers how study designs can account for heterogeneous phenotypes by adopting representative sampling schemes for recruiting the study population and making careful choices about the control population. Advantages and limitations of 16S profiling and whole-genome sequencing, the two primary techniques for measuring the microbiome, are discussed followed by an overview of bioinformatic processing of high-throughput sequencing data from these measurements. Practical insights into the downstream statistical analyses including data processing and integration, variable transformations, and data exploration are provided. The merits of regularization and ensemble modeling for analyzing microbiome data are discussed along with a recommendation for selecting modeling approaches based on data-driven simulations and objective evaluation. The review builds on several recent discussions of study design issues in microbiome research but with a stronger emphasis on the downstream and often-ignored aspects of statistical analyses that are crucial for bridging the gap between basic science and translation.


Centrosome positioning in polarized cells: common themes and variations.
The centrosome position is tightly regulated during the cell cycle and during differentiated cellular functions. Because centrosome organizes the microtubule network to coordinate both intracellular organization and cell signaling, centrosome positioning is crucial to determine either the axis of cell division, the direction of cell migration or the polarized immune response of lymphocytes. Since alteration of centrosome positioning seems to promote cell transformation and tumor spreading, the molecular mechanisms controlling centrosome movement in response to extracellular and intracellular cues are under intense investigation. Evolutionary conserved pathways involving polarity proteins and cytoskeletal rearrangements are emerging as common regulators of centrosome positioning in a wide variety of cellular contexts.


Effects of age-related gait changes on the biomechanics of slips and falls.
A laboratory study was conducted to examine gait changes associated with aging and the effect of these changes on initiation of slips and frequency of falls utilizing newly defined biomechanical parameters of slips and falls. Twenty-eight participants from two age groups (young and old) walked around a circular track at a comfortable pace wearing a safety harness. A slippery floor surface was placed on the walking track over the force plate at random time intervals without the participants' awareness. Synchronized kinetic and kinematic measurements were obtained on both slippery and non-slippery walking surfaces. The results indicated that older participants' horizontal heel contact velocity was significantly faster, step length was significantly shorter, and transitional acceleration of the whole body centre-of-mass (COM) was significantly slower than younger participants. Older participants' initial friction demand, as measured by required coefficient of friction (RCOF), was not significantly different than their younger counterparts. Additionally, older participants slipped longer and faster, and fell more often than younger participants. A comparison of horizontal heel contact velocity for participants who fell with participants who did not fall indicated that, in general, fallers' horizontal heel contact velocity was faster than non-fallers. However, a comparison of RCOF for participants who fell with participants who did not fall suggested that RCOF was not a totally deterministic factor influencing actual fall events. These findings suggest that gait changes associated with aging (especially higher horizontal heel contact velocity and slower transition of the whole body COM) affect initiation of slip-induced falls.


Seasonal prevalence of ticks and their association with dermatophilosis in cattle on the Accra plains of Ghana.
The seasonal abundance of adult ticks on cattle and their association with dermatophilosis were investigated in five herds on the coastal plains of Ghana over a 26-month period. Four genera, Amblyomma, Boophilus, Rhipicephalus and Hyalomma were identified, A. variegatum being the predominant species occurring throughout the year with two peaks of infestation, one in April-May and the other in November. A significant positive correlation was revealed between A. variegatum and dermatophilosis in four of the five herds. Significant positive correlations were found between H. m. rufipes and dermatophilosis in two of the herds and between Rh. senegalensis and dermatophilosis in one herd. Negative correlations of statistical significance were observed between Boophilus species and dermatophilosis in three of the herds. Nevertheless, it was considered that A. variegatum was the most important tick factor involved in the pathogenesis of the disease.


Alteration in angiogenic and anti-angiogenic forms of vascular endothelial growth factor-A in skeletal muscle of patients with intermittent claudication following exercise training.
The aims of this study were twofold: (1) to identify whether peripheral artery disease (PAD) patients had increased muscle concentration of angiogenic VEGF-A, anti-angiogenic VEGF₁₆₅b or VEGF receptor 1 (VEGF-R1) when compared with control subjects, and (2) to evaluate whether exercise training in PAD patients was associated with changes in muscle concentration of VEGF-A, VEGF₁₆₅b or VEGF-R1. At baseline, 22 PAD and 30 control subjects underwent gastrocnemius muscle biopsy. Twelve PAD patients were treated with supervised exercise training (SET) and underwent muscle biopsy after 3 weeks and 12 weeks of training and had sufficient tissue to measure VEGF-A, VEGF₁₆₅b and VEGF-R1 concentrations in skeletal muscle lysates by ELISA. Muscle concentrations of VEGF-A and VEGF₁₆₅b were similar in PAD patients versus controls at baseline. At both time points after the start of SET, VEGF-A levels decreased and there was a trend towards increased VEGF₁₆₅b concentrations. At baseline, VEGF-R1 concentrations were lower in PAD patients when compared with controls but did not change after SET. Skeletal muscle concentrations of VEGF-A are not different in PAD patients when compared with controls at baseline. SET is associated with a significant reduction in VEGF-A levels and a trend towards increased VEGF₁₆₅b levels. These somewhat unexpected findings suggest that further investigation into the mechanism of vascular responses to exercise training in PAD patients is warranted.


Estimating a preference-based index for a 5-dimensional health state classification for asthma derived from the asthma quality of life questionnaire.
This article presents a valuation study to estimate a preference-based index for a 5-dimensional health state classification for asthma (AQL-5D) derived from the Asthma Quality of Life Questionnaire (AQLQ). A sample of 307 members of the UK general population valued 99 asthma health states selected from the AQL-5D using the time tradeoff technique. Models were estimated to predict all possible 3125 health states defined by the AQL-5D, and the models were compared in terms of their ability to predict mean values for the 99 states. Mean health state values ranged from 0.39 to 0.94 based on an average of 22 valuations per state. A main effects model estimated on mean health state values and adjusted for consistency had the best predictive ability (mean absolute error of 0.047 and only 9/98 states with errors >0.1) and the most logical consistency with levels of the AQL-5D. The low number of valuations per state may have resulted in unreliable estimates for the models. Preference-based condition specific measures are limited in their ability to make cross-disease comparisons. This is the first study to derive a condition-specific preference-based measure from an existing measure of health-related quality of life in asthma for use in economic evaluation.


Triggers of tree mortality under drought.
Severe droughts have caused widespread tree mortality across many forest biomes with profound effects on the function of ecosystems and carbon balance. Climate change is expected to intensify regional-scale droughts, focusing attention on the physiological basis of drought-induced tree mortality. Recent work has shown that catastrophic failure of the plant hydraulic system is a principal mechanism involved in extensive crown death and tree mortality during drought, but the multi-dimensional response of trees to desiccation is complex. Here we focus on the current understanding of tree hydraulic performance under drought, the identification of physiological thresholds that precipitate mortality and the mechanisms of recovery after drought. Building on this, we discuss the potential application of hydraulic thresholds to process-based models that predict mortality.


Ethnobotany and ethnopharmacology--interdisciplinary links with the historical sciences.
In this paper we use three disparate examples to highlight the relevance of historical methods in the context of ethnobotany and ethnopharmacology. Unfortunately, in ethnopharmacology we have only very few examples where such historical depth is possible. On the other hand the first hand data available through ethnopharmacological research, may be of relevance for interpreting historical information. Three distinct methods were used successfully in the retrospective exploration of diachronic data. In order to gain an insight into historical developments in the use of plant species and evolution of pharmacopoeias we used a botanico-historical approach (Rosmarinus officinalis), one that combines linguistic and statistical methods (Popoluca/Mixe), and one that uses historical documentary evidence (Ch'orti). We hope that this methodological discussion encourages a wider use of such historical methods in ethnopharmacology and related areas of research.


High-dose therapy and autologous stem cell transplant in older adults with multiple myeloma.
Randomized trials showing that high-dose therapy with autologous stem cell transplant (ASCT) improved the overall survival (OS) in multiple myeloma (MM) excluded patients over age 65. To compare the outcomes of older adults with MM who underwent ASCT with non-transplant strategies, we identified 146 patients aged 65-77 with newly diagnosed MM seen in the Washington University School of Medicine from 2000 to 2010. Survival among patients who did (N=62) versus did not (N=84) undergo ASCT was compared using Cox proportional hazards modeling, controlling for comorbidities, Eastern Cooperative Oncology Group performance status (PS) and the propensity to undergo ASCT. Median age was 68 years (range 65-77). PS and comorbidities did not differ significantly between those who did versus those who did not undergo ASCT. Median OS was significantly longer in patients who underwent ASCT than in those who did not (median 56.0 months (95% confidence intervals (CIs) 49.1-65.4) versus 33.1 months (24.3-43.1), P=0.004). Adjusting for PS, comorbidities, Durie-Salmon stage and the propensity to undergo ASCT, ASCT was associated with superior OS (HR for mortality 0.52 (95% CI 0.30-0.91), P=0.02). In a cohort of older adults with MM, undergoing ASCT was associated with a nearly 50% lower mortality, after controlling for PS, comorbidities, stage and the propensity to undergo ASCT.