Critical care nurses' knowledge and attitudes and their perspectives toward promoting advance directives and end-of-life care.
End-of-life care can be a difficult and challenging process for critical care nurses in intensive care units (ICUs) due to the care plan shifts from providing life-sustaining measures to end-of-life care. The aims of this study were to assess critical care nurses' perceived knowledge and attitudes toward end-of-life care, as well as their perspectives on promoting advance directives and the associated factors. A cross-sectional study was undertaken in an acute major metropolitan medical center in northern Taiwan between February and March 2020, and 250 critical care nurses were invited to participate in the study. Data on demographics, self-perceived knowledge of end-of-life care, attitudes toward end-of-life care, and perspectives of promoting advance directives were collected. A multiple linear regression model with stepwise selection was used to identify factors associated with their perspectives of promoting advance directives. The law related to end-of-life care was rated as the least familiar part of the self-perceived end-of-life care knowledge, while 'I have sufficient knowledge to care for patients who have accepted end-of-life care' was the lowest level of agreement in attitude scores among critical care nurses. Increased levels of perceived knowledge (β = 0.134; p = 0.045) and attitudes (β = 0.423; p < 0.001) toward end-of-life care were associated with the perspectives of promoting advance directives. Nurses who worked in cardiac (β = -0.234; p < 0.001) and respiratory ICUs (β = -0.135; p = 0.024) had less motivation to promote advance directives (F = 16.943; p < 0.001). Given their important contributions to ICU care services, appropriate and meaningful support is required to optimize critical care nurses' involvement in end-of-life care. This study demonstrated a significant impact on perspectives of promoting advance directives of critical care nurse participants. Findings from this study can inform the design of effective nurse support programs to enhance the promotion of advance directives in intensive care settings.


Validity of a self-reported diagnosis of depression among participants in a cohort study using the Structured Clinical Interview for DSM-IV (SCID-I).
Depression assessment in population studies is usually based on depressive symptoms scales. However, the use of scales could lead to the choice of an arbitrary cut-off point depending on the sample characteristics and on the patient diagnosis. Thus, the use of a medical diagnosis of depression could be a more appropriate approach. To validate a self-reported physician diagnosis of depression using the Structured Clinical Interview for DSM-IV (SCID-I) as Gold Standard and to assess the factors associated to a valid self-reported diagnosis. The SUN Project is a cohort study based on university graduates followed-up through postal questionnaires. The response to the question included in the questionnaire: Have you ever been diagnosed of depression by a physician? was compared to that obtained through the SCID-I applied by a psychiatrist or a clinical psychologist. The percentages of confirmed depression and non-depression were assessed for the overall sample and according to several characteristics. Logistic regression models were fitted to ascertain the association between different factors and a correct classification regarding depression status. The percentage of confirmed depression was 74.2%; 95% confidence interval (95% CI) = 63.3-85.1. Out of 42 participants who did not report a depression diagnosis in the questionnaire, 34 were free of the disease (%confirmed non-depression = 81.1%; 95% CI = 69.1-92.9). The probability of being a true positive was higher among ex-smokers and non-smokers and among those overweight or obese but the differences were not statistically significant. The validity of a self-reported diagnosis of depression in the SUN cohort is adequate. Thus, this question about depression diagnosis could be used in further investigations regarding this disease in this graduate cohort study.


Genetics in arterial calcification: pieces of a puzzle and cogs in a wheel.
Artery calcification reflects an admixture of factors such as ectopic osteochondral differentiation with primary host pathological conditions. We review how genetic factors, as identified by human genome-wide association studies, and incomplete correlations with various mouse studies, including knockout and strain analyses, fit into "pieces of the puzzle" in intimal calcification in human atherosclerosis, and artery tunica media calcification in aging, diabetes mellitus, and chronic kidney disease. We also describe in sharp contrast how ENPP1, CD73, and ABCC6 serve as "cogs in a wheel" of arterial calcification. Specifically, each is a minor component in the function of a much larger network of factors that exert balanced effects to promote and suppress arterial calcification. For the network to normally suppress spontaneous arterial calcification, the "cogs" ENPP1, CD73, and ABCC6 must be present and in working order. Monogenic ENPP1, CD73, and ABCC6 deficiencies each drive a molecular pathophysiology of closely related but phenotypically different diseases (generalized arterial calcification of infancy (GACI), pseudoxanthoma elasticum (PXE) and arterial calcification caused by CD73 deficiency (ACDC)), in which premature onset arterial calcification is a prominent but not the sole feature.


The characterization of Salmonella enterica serotypes isolated from the scalder tank water of a commercial poultry processing plant: Recovery of a multidrug-resistant Heidelberg strain.
The recent multistate outbreak of a multidrug-resistant (MDR) Salmonella Heidelberg strain from commercial poultry production highlights the need to better understand the reservoirs of these zoonotic pathogens within the commercial poultry production and processing environment. As part of a larger study looking at temporal changes in microbial communities within the major water tanks within a commercial processing facility, this paper identifies and characterizes Salmonella enterica isolated from the water in a final scalder tank at 3 times during a typical processing day: prior to the birds entering the tank (start), halfway through the processing day (mid), and after the final birds were scalded (end). Over 3 consecutive processing days, no Salmonella were recovered from start-of-day water samples, while a total of 56 Salmonella isolates were recovered from the mid-day and end-of-day scalder water samples. Traditional and newer PCR-based serotyping methods eventually identified these isolates as either group C3 S. Kentucky (n=45) and group B S. Heidelberg (n=11). While none of the S. Kentucky isolates possessed any resistances to the antimicrobials tested, all S. Heidelberg isolates were found to be multidrug resistant to 5 specific antimicrobials representing 3 antimicrobial classes. Due to the potential public health impact of S. Heidelberg and the recent nationwide poultry-associated outbreak of multidrug-resistant S. Heidelberg, future studies should focus on understanding the transmission and environmental growth dynamics of this serotype within the commercial poultry processing plant environment.


Grazing optimization, nutrient cycling, and spatial heterogeneity of plant-herbivore interactions: should a palatable plant evolve?
Can the evolution of plant defense lead to an optimal primary production? In a general theoretical model, Loreau (1995) and de Mazancourt et al. (1998, 1999) have shown that herbivory could increase primary production up to a moderate rate of grazing intensity through recycling of a limiting nutrient, provided several conditions are fulfilled. In the present paper, we assume: (1) grazing intensity is controlled by plants through their level of palatability; and (2) plant fitness is determined by its productivity. We explore the conditions under which such an optimal production may be reached through natural selection. We model two competing plant types that differ only in palatability and are distributed in a patchy landscape determined by the plant-herbivore interaction. Patch size is determined by herbivore behavior: herbivores recycle nutrient homogeneously within patches, but recycle nutrient proportionally to consumption between patches. The model shows that a strategy of intermediate palatability can be adaptive in response to a small herbivore that lives on and recycles nutrient around one or a few individual plants. For moderately small herbivores, plant palatability may evolve towards one of two local convergent strategies, depending on the initial conditions. For medium- to large-sized herbivores, the nonpalatable strategy is always selected. We discuss the functional and evolutionary implications of these results, and suggest that the traditional dichotomy describing antagonistic and mutualistic interactions may be misleading.


A novel MATE family efflux pump contributes to the reduced susceptibility of laboratory-derived Staphylococcus aureus mutants to tigecycline.
Tigecycline, an expanded-broad-spectrum glycylcycline antibiotic is not affected by the classical tetracycline resistance determinants found in Staphylococcus aureus. The in vitro selection of mutants with reduced susceptibility to tigecycline was evaluated for two methicillin-resistant S. aureus strains by serial passage in increasing concentrations of tigecycline. Both strains showed a stepwise elevation in tigecycline MIC over a period of 16 days, resulting in an increase in tigecycline MIC of 16- and 32-fold for N315 and Mu3, respectively. Transcriptional profiling revealed that both mutants exhibited over 100-fold increased expression of a gene cluster, mepRAB (multidrug export protein), encoding a MarR-like transcriptional regulator (mepR), a novel MATE family efflux pump (mepA), and a hypothetical protein of unknown function (mepB). Sequencing of the mepR gene in the mutant strains identified changes that presumably inactivated the MepR protein, which suggested that MepR functions as a repressor of mepA. Overexpression of mepA in a wild-type background caused a decrease in susceptibility to tigecycline and other substrates for MATE-type efflux pumps, although it was not sufficient to confer high-level resistance to tigecycline. Complementation of the mepR defect by overexpressing a wild-type mepR gene reduced mepA transcription and lowered the tigecycline MIC in the mutants. Transcription of tet(M) also increased by over 40-fold in the Mu3 mutant. This was attributed to a deletion in the promoter region of the gene that removed a stem-loop responsible for transcriptional attenuation. However, overexpression of the tet(M) transcript in a tigecycline-susceptible strain was not enough to significantly increase the MIC of tigecycline. These results suggest that the overexpression of mepA but not tet(M) may contribute to decreased susceptibility of tigecycline in S. aureus.


High Glucose Variability in Hospitalized Patients with Type 1 Diabetes Mellitus.
Glucose variability (GV) has been increasingly (or more extensively) studied in patients with type 1 diabetes (T1D) in the ambulatory setting; limited data exist on GV in hospitalized patients with T1D. Retrospective single center cohort study, we analyzed in-hospital glucose measurements to assess GV in 736 hospitalized patients in different units over a consecutive 5-year period of time. GV was assessed by mean blood glucose (BG), Average daily risk range (ADRR), high BG index, and low BG index. To place our findings in context, we conducted a systematic review using Cochrane collaboration methodology to critically analyze current published literature on GV in hospitalized T1D patients. Overall, glycemic control was suboptimal with mean BG 183 ± 51.5 mg/dL and mean ADRR 35 with only 16% patients being categorized as low risk (ADRR <20) for hypo or hyperglycemia. Patients admitted in medical units had mean BG of 194.4 ± 42.8 mg/dL (95% CI = 101.2-346.6) and ADRR of 39.4 ± 16 (95% CI = 1.3-118.7), which were higher than the patients admitted in the surgical units (mean BG 168.1 ± 35.7 (95% CI = 74.8-301.8) and mean ADRR (28.8 ± 13.1 [95% CI = 0.3-93.1]). For the systematic review, initial search yielded 2336 studies for screening, however, none of them had data specific for T1D. GV is high in hospitalized T1D patients admitted at our tertiary care center. Review of the literature shows paucity of data regarding GV in hospitalized patients with T1D.


Mediators of Change in a Parent Training Program for Early ADHD Difficulties: The Role of Parental Strategies, Parental Self-Efficacy, and Therapeutic Alliance.
Objective: The aim was to explore mediators of change in parent training (PT) for 3- to 8-year-old children with ADHD difficulties. Method: Parents of 64 children received PT with Incredible Years® and assessed child ADHD symptoms and conduct problems and their parenting strategies, parental self-efficacy, and therapeutic alliance before, during, and after PT. Product-of-coefficients mediation analyses in multilevel models were applied, and causal relations between mediators and outcome were investigated in time-lagged analyses. Results: Increased parental self-efficacy and reduced negative parenting statistically mediated reductions in ADHD and conduct problems in the product-of-coefficient analyses. However, time-lagged analyses were unable to detect a causal relation between prior change in mediators and subsequent child symptom reduction. There was limited evidence of therapeutic alliance as mediator of child symptom reduction or change in parenting variables. Conclusion: Parental self-efficacy and reductions in negative parenting may mediate change in PT, but more fine-grained time-lagged analyses are needed to establish causality.


The validity, reliability and normative scores of the parent, teacher and self report versions of the Strengths and Difficulties Questionnaire in China.
The Strengths and Difficulties Questionnaire (SDQ) has become one of the most widely used measurement tools in child and adolescent mental health work across the globe. The SDQ was originally developed and validated within the UK and whilst its reliability and validity have been replicated in several countries important cross cultural issues have been raised. We describe normative data, reliability and validity of the Chinese translation of the SDQ (parent, teacher and self report versions) in a large group of children from Shanghai. The SDQ was administered to the parents and teachers of students from 12 of Shanghai's 19 districts, aged between 3 and 17 years old, and to those young people aged between 11 and 17 years. Retest data was collected from parents and teachers for 45 students six weeks later. Data was analysed to describe normative scores, bandings and cut-offs for normal, borderline and abnormal scores. Reliability was assessed from analyses of internal consistency, inter-rater agreement, and temporal stability. Structural validity, convergent and discriminant validity were assessed. Full parent and teacher data was available for 1965 subjects and self report data for 690 subjects. Normative data for this Chinese urban population with bandings and cut-offs for borderline and abnormal scores are described. Principle components analysis indicates partial agreement with the original five factored subscale structure however this appears to hold more strongly for the Prosocial Behaviour, Hyperactivity - Inattention and Emotional Symptoms subscales than for Conduct Problems and Peer Problems. Internal consistency as measured by Cronbach's alpha coefficient were generally low ranging between 0.30 and 0.83 with only parent and teacher Hyperactivity - Inattention and teacher Prosocial Behaviour subscales having alpha > 0.7. Inter-rater correlations were similar to those reported previously (range 0.23 - 0.49) whilst test retest reliability was generally lower than would be expected (range 0.40 - 0.79). Convergent and discriminant validity are supported. We report mixed findings with respect the psychometric properties of the Chinese translation of the SDQ. Reliability is a particular concern particularly for Peer Problems and self ratings by adolescents. There is good support for convergent validity but only partial support for structural validity. It may be possible to resolve some of these issues by carefully examining the wording and meaning of some of the current questions.


Treatment of fistulas in ano with fibrin glue.
Recent publications show promising results using fibrin glue in the treatment of anal fistulas. The technique is simple, repeatable, with minor surgical trauma so that sphincter function is preserved and further treatment options are not compromised. The aim of this pilot study was to analyse if we could reproduce the results reported in the literature, using this simple technique. Patients with a primary or recurrent anal fistula were included in this trial. Patients with a complex fistula were excluded. Under general or spinal anaesthesia, the fistulas were curetted and injected with fibrin glue. Follow-up visits were scheduled for 1 week, 6 weeks and 6 months. Twenty-seven patients were included. The overall success rate was 33% after a mean follow-up of 27 weeks. Patients with a recurrent fistula had a poorer outcome (success rate 14%). None of the patients suffered from postoperative continence problems, and no other complications were recorded. This study confirms the safety of fibrin glue in the treatment of anal fistulas. However, a high success rate could not be reproduced.


The contribution of posttraumatic stress disorder and mild traumatic brain injury to persistent post concussive symptoms following motor vehicle accidents.
Persistent postconcussive symptoms (PPCS) are a set of physical, cognitive, emotional, and behavioral symptoms that often follow mild traumatic brain injury (mTBI). Some of these symptoms also occur in posttraumatic stress disorder (PTSD). The current study examined the unique contribution of mTBI and PTSD to PPCS. The roles of neurocognitive and motivational factors were also addressed. Sixty one children and adolescents (ages 6-18), at least 3 months post motor vehicle accident (MVA), participated in the study. All participants were diagnosed with PTSD symptoms. Thirty three participants met mTBI criteria, and 28 did not. Standard instruments for assessment included a semistructured clinical interview, self-report questionnaires, and a neuropsychological evaluation. No differences were found between the mTBI and non-TBI groups on any of the emotional or neurocognitive measures, including PPCS symptoms. Multiple regression analyses revealed that emotional status, such as state anxiety and depression, were the best predictors of PPCS. Furthermore, hierarchical regression analyses revealed a double mediation model, in which suboptimal effort mediated the relationship between neurocognitive performance and PPCS, and emotional status mediated the relationship between suboptimal effort and PPCS. These findings underscore the importance of emotional status in the diagnosis of PPCS among children who suffer from PTSD. It is possible that PPCS reflect a more general expression of accident-related emotional distress, rather than being a direct result of the injury. (PsycINFO Database Record


Atypical and malignant solitary fibrous tumors in extrathoracic locations: evidence of their comparability to intra-thoracic tumors.
Solitary fibrous tumor (SFT), first described as a pleural lesion, has been reported at numerous extrathoracic sites over the past 10 years. About 10% to 15% of intrathoracic SFTs are histologically or clinically malignant, but such cases have very rarely been described at other locations. Among 92 cases of extrathoracic SFT in our files, we identified 10 that either had recurred (2 cases) or had a least one atypical histologic feature (8 cases). The ten tumors occurred in five men and five women, 32 to 81 years old (median 56), measured 1.9 cm to 20 cm (median 11.5 cm), and were located in the abdomen/pelvis (4 cases), retroperitoneum (3 cases), groin, trunk, and upper arm. Nuclear atypia (8 cases), markedly increased cellularity (6 cases), areas of necrosis (4 cases), and greater than 4 mitoses/10 HPFs (3 cases) were seen in addition to the typical histologic features of SFT. Six tumors had at least two of these atypical histologic features. Nine cases were positive for CD34, six were positive for O-13, and one was focally positive for smooth muscle actin. Eight were excised completely. Subsequent follow-up revealed tumor relapse in eight cases (follow up 6-180 months, median 24). Four patients had local recurrence at 12 to 168 months. Distant metastasis developed at 1 to 6 years in five cases with spread to lung (2 cases), liver (4 cases), and bone. Metastasis or local recurrence developed within 2 years in five patients. To date, no patient has died of their tumor. These findings demonstrate that nuclear atypia, hypercellularity, greater than 4 mitoses/10 HPFs, and necrosis may be seen in up to 10% of extrathoracic SFTs, and are associated with, but are not by themselves predictive of, aggressive clinical behavior. In addition, our findings confirm that the behavior of extrathoracic SFTs is unpredictable, entirely comparable to that of their better known pleural counterparts, and confirm that patients with SFTs in all locations require careful, long-term follow up. It is probably unwise to regard any such lesion as definitely benign.


Retigabine: chemical synthesis to clinical application.
Retigabine [D23129; N-(2-amino-4-(4-fluorobenzylamino)-phenyl)carbamic acid ethyl ester] is an antiepileptic drug with a recently described novel mechanism of action that involves opening of neuronal K(V)7.2-7.5 (formerly KCNQ2-5) voltage-activated K(+) channels. These channels (primarily K(V)7.2/7.3) enable generation of the M-current, a subthreshold K(+) current that serves to stabilize the membrane potential and control neuronal excitability. In this regard, retigabine has been shown to have a broad-spectrum of activity in animal models of electrically-induced (amygdala-kindling, maximal electroshock) and chemically-induced (pentylenetetrazole, picrotoxin, NMDA) epileptic seizures. These encouraging results suggest that retigabine may also prove useful in the treatment of other diseases associated with neuronal hyperexcitability. Neuropathic pain conditions are characterized by pathological changes in sensory pathways, which favor action potential generation and enhanced pain transmission. Although sometimes difficult to treat with conventional analgesics, antiepileptics can relieve some symptoms of neuropathic pain. A number of recent studies have reported that retigabine can relieve pain-like behaviors (hyperalgesia and allodynia) in animal models of neuropathic pain. Neuronal activation within several key structures within the CNS can also be observed in various animal models of anxiety. Moreover, amygdala-kindled rats, which have a lowered threshold for neuronal activation, also display enhanced anxiety-like responses. Retigabine dose-dependently reduces unconditioned anxiety-like behaviors when assessed in the mouse marble burying test and zero maze. Early clinical studies have indicated that retigabine is rapidly absorbed and distributed, and is resistant to first pass metabolism. Tolerability is good in humans when titrated up to its therapeutic dose range (600-1200 mg/day). No tolerance, dependence or withdrawal potential has been reported, although adverse effects can include mild dizziness, headache, nausea and somnolence. Thus, retigabine may prove to be useful in the treatment of a diverse range of disease states in which neuronal hyperexcitability is a common underlying factor.


Bacterial gene inserted in an engineered RNA virus: efficient expression in monocotyledonous plant cells.
Brome mosaic virus (BMV) is a plant virus whose genome consists of three RNA components. A previously described viral complementary DNA expression system has been used to express both wild-type and altered genomic RNA's in barley protoplasts. Variants of BMV RNA3 were constructed in which the coat gene had been removed or replaced with sequences encoding chloramphenicol acetyltransferase (CAT). CAT sequences were also inserted near the 5' end of the intact coat gene. When inoculated on protoplasts together with transcripts of BMV RNA's 1 and 2, all of these RNA3 derivatives were replicated and produced subgenomic RNA's analogous to the normal subgenomic coat protein messenger RNA. RNA3 derivatives in which the CAT coding sequences were oriented with the same polarity as viral genes produced significant CAT activity in protoplasts. CAT expression was improved by inserting the CAT gene in frame with the upstream coat protein initiation codon, and exceeded expression in plant cells transformed with Ti plasmid-based vectors.


Interpreting top-down mass spectra using spectral alignment.
Recent advances in mass spectrometry instrumentation, such as FTICR and OrbiTrap, have made it possible to generate high-resolution spectra of entire proteins. While these methods offer new opportunities for performing "top-down" studies of proteins, the computational tools for analyzing top-down data are still scarce. In this paper we investigate the application of spectral alignment to the problem of identifying protein forms in top-down mass spectra (i.e., identifying the modifications, mutations, insertions, and deletions). We demonstrate how spectral alignment efficiently discovers protein forms even in the presence of numerous modifications and how the algorithm can be extended to discover positional isomers from spectra of mixtures of isobaric protein forms.


Systematic investigation of initiatives to reduce seclusion and restraint in a state psychiatric hospital.
This study used an experimental design to examine the effect of systematic implementation of behavioral interventions on the rate of seclusion and restraint in an inpatient psychiatric hospital. With a variant of the multiple-baseline design, a model designed to reduce seclusion and restraint was implemented at a large state-funded hospital in the southeastern United States. The implementation schedule was established such that each of five inpatient units was randomly assigned to implement the intervention components in a different order, and each unit served as its own control. Participants were patients and staff, for a total of 89,783 patient-days over a 3.5-year period from January 2005 through June 2008. The components included trauma-informed care training, changes to unit rules and language, changes to the physical characteristics of the therapeutic environment, and involvement of patients in treatment planning. The rate of inpatient psychiatric seclusion and restraint (per patient day) was tracked continuously during the 3.5-year period. A significant reduction of 82.3% (p=.008) in the rate of seclusion and restraint was observed between the baseline phase (January 2005 through February 2006) and the follow-up, postintervention phase (April 2008 through June 2008). After control for illness severity and nonspecific effects associated with an observation-only phase, changes to the physical environment were uniquely associated with a significant reduction in rate of seclusion and restraint during the intervention rollout period. These data suggest that substantial reductions in use of seclusion and restraint are possible in inpatient psychiatric settings and that changes to the physical characteristics of the therapeutic environment may have a significant effect on use of seclusion and restraint.


Multiple double cross-section transmission electron microscope sample preparation of specific sub-10 nm diameter Si nanowire devices.
The ability to prepare multiple cross-section transmission electron microscope (XTEM) samples from one XTEM sample of specific sub-10 nm features was demonstrated. Sub-10 nm diameter Si nanowire (NW) devices were initially cross-sectioned using a dual-beam focused ion beam system in a direction running parallel to the device channel. From this XTEM sample, both low- and high-resolution transmission electron microscope (TEM) images were obtained from six separate, specific site Si NW devices. The XTEM sample was then re-sectioned in four separate locations in a direction perpendicular to the device channel: 90° from the original XTEM sample direction. Three of the four XTEM samples were successfully sectioned in the gate region of the device. From these three samples, low- and high-resolution TEM images of the Si NW were taken and measurements of the NW diameters were obtained. This technique demonstrated the ability to obtain high-resolution TEM images in directions 90° from one another of multiple, specific sub-10 nm features that were spaced 1.1 μm apart.


[Verbal memory disorders provoked by a variety of stokes].
In recent years a consensus has been reached about the neuroanatomical substrate of verbal memory, but this state of knowledge has not yet been implemented in clinical practice. One reason for this may be that most of the neuroscientific studies on verbal memory used different neuropsychological instruments and that only a small set of patient groups with the same etiology but different lesion sites were analysed. Returning to three earlier studies, we analyse the possibility to make differential judgements on the verbal memory impairments of four different patient groups by using the California Verbal Learning Test (CVLT). We compare patients with left-sided (n = 16) and right-sided (n = 10) posterior cerebral artery infarcts and patients with infarcts of the left (n = 10) or right (n = 21) frontal lobe, and we integrated data about their retention errors that had not been analysed so far. Our findings reveal significant differences between these patient groups, concerning the quantitative aspects of impairments, and also the profiles of memory errors (recall, interference and perseverations). Our study documents a relation between the site of the lesion and the type of verbal memory impairment, agreeing with some of the most recent neuroscientific findings. Starting from this observation we try to define a neuropsychological pattern of memory impairment which enables differential clinical diagnoses using the CVLT as a memory test.


Microscale mechanical properties of single elastic fibers: the role of fibrillin-microfibrils.
Micromechanical properties of single elastic fibers and fibrillin-microfibrils, isolated from equine ligamentum nuchae using chemical and enzymatic methods, were determined with atomic force microscopy (AFM). Young's moduli of single elastic fibers immersed in water, devoid of or containing fibrillin-microfibrils, were determined using bending tests. Bending freely suspended elastic fibers on a micro-channeled substrate by a tip-less AFM cantilever generated a force versus displacement curve from which Young's moduli were calculated. For single elastic fibers, Young's moduli in the range of 0.3-1.5 MPa were determined, values not significantly affected by the presence of fibrillin-microfibrils. To further understand the role of fibrillin-microfibrils in vertebrate elastic fibers, layers of fibrillin-microfibrils were subjected to nano-indentation tests. From the slope of the force versus indentation curves, Young's moduli ranging between 0.56 and 0.74 MPa were calculated. The results suggest that fibrillin-microfibrils are not essential for the mechanical properties of single vertebrate elastic fibers.


People's care seeking journey for a chronic illness in rural India: Implications for policy and practice.
Drawing on interviews conducted in 2019-2020, across twenty villages in India, this paper unpacks how people with chronic illness navigate complex care-seeking terrain. We show how the act of seeking care involves navigating through personal, family, social, economic, cultural, and most importantly, difficult health systems spaces-and entails making difficult social, moral, and financial choices. We show how the absence of reliable and accessible points of first contact for primary care results in people running from pillar to post, taking wrong turns, and becoming disappointed, frustrated, and, sometimes, impoverished. We reveal the complex individual and social dynamics of hope and misplaced and misguided expectations, as well as social obligations and their performance that animate the act of navigating care in rural India. We shine light on how a health system with weak primary care and poor regulation amplifies the medical, social, and financial consequences of an otherwise manageable chronic illness, and how these consequences are the worst for those with the least social, network and economic capital. Crucially we highlight the problematic normalisation of the absence of reliable primary care services for chronic illness in India, in rural India specifically. We signpost implications for research, and for policy and practice in India and similar health system contexts, i.e. those with weak primary care and poor regulation of the private sector. We argue that in India, having in place accessible, good quality, and trustworthy sources of advice and care for chronic illness at the first point of call, for all, is critical. We contend that this first point of call should be quality, public primary care services. We conclude that if such arrangements are in place in public services, people will use them.


Defining a structural and kinetic rationale for paralogous copies of phenylacetate-CoA ligases from the cystic fibrosis pathogen Burkholderia cenocepacia J2315.
The phenylacetic acid (PAA) degradation pathway is the sole aerobic route for phenylacetic acid metabolism in bacteria and facilitates degradation of environmental pollutants such as styrene and ethylbenzene. The PAA pathway also is implicated in promoting Burkholderia cenocepacia infections in cystic fibrosis patients. Intriguingly, the first enzyme in the PAA pathway is present in two copies (paaK1 and paaK2), yet each subsequent enzyme is present in only a single copy. Furthermore, sequence divergence indicates that PaaK1 and PaaK2 form a unique subgroup within the adenylate-forming enzyme (AFE) superfamily. To establish a biochemical rationale for the existence of the PaaK paralogs in B. cenocepacia, we present high resolution x-ray crystal structures of a selenomethionine derivative of PaaK1 in complex with ATP and adenylated phenylacetate intermediate complexes of PaaK1 and PaaK2 in distinct conformations. Structural analysis reveals a novel N-terminal microdomain that may serve to recruit subsequent PAA enzymes, whereas a bifunctional role is proposed for the P-loop in stabilizing the C-terminal domain in conformation 2. The potential for different kinetic profiles was suggested by a structurally divergent extension of the aryl substrate pocket in PaaK1 relative to PaaK2. Functional characterization confirmed this prediction, with PaaK1 possessing a lower K(m) for phenylacetic acid and better able to accommodate 3' and 4' substitutions on the phenyl ring. Collectively, these results offer detailed insight into the reaction mechanism of a novel subgroup of the AFE superfamily and provide a clear biochemical rationale for the presence of paralogous copies of PaaK of B. cenocepacia.


Effect of Health-Related Uncertainty and Natural Variability on Health Impacts and Cobenefits of Climate Policy.
Climate policy can mitigate health risks attributed to intensifying air pollution under climate change. However, few studies quantify risks of illness and death, examine their contribution to climate policy benefits, or assess their robustness in light of natural climate variability. We employ an integrated modeling framework of the economy, climate, air quality, and human health to quantify the effect of natural variability on U.S. air pollution impacts under future climate and two global policies (2 and 2.5 °C stabilization scenarios) using 150 year ensemble simulations for each scenario in 2050 and 2100. Climate change yields annual premature deaths related to fine particulate matter and ozone (95CI: 25 000-120 000), heart attacks (900-9400), and lost work days (3.6M-4.9M) in 2100. It raises air pollution health risks by 20%, while policies avert these outcomes by 40-50% in 2050 and 70-88% in 2100. Natural variability introduces "climate noise", yielding some annual estimates with negative cobenefits, and others that reach 100% of annual policy costs. This "noise" is three times the magnitude of uncertainty (95CI) in health and economic responses in 2050. Averaging five annual simulations reduces this factor to two, which is still substantially larger than health-related uncertainty. This study quantifies the potential for inaccuracy in climate impacts projected using too few annual simulations.


Relationship between location and activity in injurious falls: an exploratory study.
Knowledge about the circumstances under which injurious falls occur could provide healthcare workers with better tools to prevent falls and fall-related injuries. Therefore, we assessed whether older persons who sustain an injurious fall can be classified into specific fall types, based on a combination of fall location and activity up to the moment of the fall. In addition, we assessed whether specific injurious fall types are related to causes of the fall, consequences of the fall, socio-demographic characteristics, and health-related characteristics. An exploratory, cross-sectional study design was used to identify injurious fall types. The study population comprised 333 community-dwelling Dutch elderly people aged 65 years or over who attended an accident and emergency department after a fall. All participants received a self-administered questionnaire after being discharged home. The questionnaire comprised items concerning circumstances of the injurious fall, causes of the fall, consequences of the fall, socio-demographic characteristics and health-related characteristics. Injurious fall types were distinguished by analyzing data by means of HOMALS (homogeneity analysis by means of alternating least squares). We identified 4 injurious fall types: 1) Indoor falls related to lavatory visits (hall and bathroom); 2) Indoor falls during other activities of daily living; 3) Outdoor falls near the home during instrumental activities of daily living; 4) Outdoor falls away from home, occurring during walking, cycling, and shopping for groceries. These injurious fall types were significantly related to age, cause of the fall, activity avoidance and daily functioning. The face validity of the injurious fall typology is obvious. However, we found no relationship between the injurious fall types and severity of the consequences of the fall. Nevertheless, there appears to be a difference between the prevalence of fractures and the cause of the fall between the injurious fall types. Our data suggests that with regard to prevention of serious injuries, we should pay special attention to outdoor fallers and indoor fallers during lavatory visits. In addition, we should have special attention for causes of the fall. However, the conclusions reached in this exploratory analysis are tentative and need to be validated in a separate dataset.


Circular dichroism spectra and electrophoretic mobility shift assays show that human replication protein A binds and melts intramolecular G-quadruplex structures.
Noncanonical DNA structures such as G-quadruplexes might obstruct the binding of hRPA, compromising the accuracy of replication, and be a source of genomic instability. In this study, circular dichroism (CD) and electrophoretic mobility shift assay (EMSA) experiments were used to show that hRPA can bind and melt nontelomeric, intramolecular DNA G-quadruplexes under physiologically germane conditions. EMSA results show that hRPA binds to a 58-mer that includes an embedded quadruplex with an affinity equal to or greater than to nonquadruplex forming 58-mers. Moreover, hRPA binds to a 26-mer purine-rich quadruplex-forming sequence with an affinity indistinguishable from that for binding to the complementary pyrimidine-rich sequence. Under the same conditions, hRPA does not have significant affinity for binding to the duplex formed from the two sequences. Thus, DNA secondary structures can significantly modulate the binding affinity of hRPA over and above its known preference for pyrimidine-rich single-stranded sequences, so that at least some intramolecular G-quadruplex structures may not inhibit hRPA binding during DNA replication. CD spectral changes in combination with EMSA titrations suggest that one hRPA heterotrimer is sufficient to form a stable complex with an unfolded 26-mer G-quadruplex prior to the binding of a second hRPA molecule.


Microtensile bond strength of bulk-fill restorative composites to dentin.
To facilitate the easier placement of direct resin composite in deeper cavities, bulk fill composites have been introduced. The Mechanical stability of fillings in stress bearing areas restored with bulk-fill resin composites is still open to question, since long term clinical studies are not available so far. Thus, the objective of the study was to evaluate and compare the microtensile bond strength of three bulk-fill restorative composites with a nanohybrid composite. Class I cavities were prepared on sixty extracted mandibular molars. Teeth were divided into 4 groups (n= 15 each) and in group I, the prepared cavities were restored with nanohybrid (Filtek Z250 XT) restorative composite in an incremental manner. In group II, III and IV, the bulk-fill composites (Filtek, Tetric EvoCeram, X-tra fil bulk-fill restoratives) were placed as a 4 mm single increment and light cured. The restored teeth were subjected to thermocycling and bond strength testing was done using instron testing machine. The mode of failure was assessed by scanning electron microscope (SEM). The bond strength values obtained in megapascals (MPa) were subjected to statistical analysis, using SPSS/PC version 20 software.One-way ANOVA was used for groupwise comparison of the bond strength. Tukey's Post Hoc test was used for pairwise comparisons among the groups. The highest mean bond strength was achieved with Filtek bulk-fill restorative showing statistically significant difference with Tetric EvoCeram bulk-fill (p< 0.003) and X-tra fil bulk-fill (p<0.001) composites. Adhesive failures are mostly observed with X-tra fil bulk fill composites, whereas mixed failures are more common with other bulk fill composites. Bulk-fill composites exhibited adequate bond strength to dentin and can be considered as restorative material of choice in posterior stress bearing areas. Key words:Bond strength, Bulk-fill restoratives, Configuration factor, Polymerization shrinkage.


Frequency and mechanism of daytime pulmonary hypertension in patients with obstructive sleep apnoea syndrome.
In order to study the frequency and the mechanisms of daytime pulmonary hypertension (PH) in obstructive sleep apnoea syndrome (OSAS) lung function tests, blood gas analysis and right-heart catheterization were performed in 46 consecutive patients. OSAS was assessed by polysomnography. 9 patients only (20%) had PH (mean pulmonary artery pressure (Ppa) greater than or equal to 20 mmHg). Patients with PH had lower daytime PaO2 (60.8 +/- 7.6 vs. 76.2 +/- 9.4 mmHg; p less than 0.001), higher daytime PaCO2 (44.8 +/- 4.2 vs. 38.0 +/- 4.0 mmHg; p less than 0.001), lower forced vital capacity (FVC) and forced expiratory volume (FEV1) (p less than 0.001), but the severity of OSAS was not different whether PH was present or not (apnoea index: 62 +/- 34 hour in the PH group vs. 65 +/- 40 hour, apnoea + hypopnoea index 102 +/- 33 hour in the PH group vs. 86 +/- 36 hour, lowest sleep SaO2: 59 +/- 21% in the PH group vs. 66 +/- 18%). There were significant correlations between Ppa and: daytime PaO2 (r = -0.61; p less than 0.001), PaCO2 (r = 0.55; p less than 0.001), FEV1 (r = -0.52; p less than 0.001) but not between Ppa and apnoea index, apnoea + hypopnoea index, lowest sleep SaO2. PH and daytime hypoxaemia were associated either with chronic airway obstruction or with severe obesity.


Cost-effectiveness of human papillomavirus vaccination and cervical cancer screening in Thailand.
To assess the health and economic outcomes of various screening and vaccination strategies for cervical cancer prevention. Cost-effectiveness analysis from a societal perspective. Thailand. Females aged 9 years and older. Using a mathematical model of human papillomavirus (HPV) infection and cervical cancer, calibrated to epidemiological data from Thailand, we estimated the cost-effectiveness of pre-adolescent HPV vaccination, screening [visual inspection with acetic acid (VIA), HPV DNA testing, and cytology] between one and five times per lifetime in adulthood, and combined pre-adolescent vaccination and screening. Vaccine efficacy, coverage, cost, and screening frequency were varied in sensitivity analyses. Incremental cost-effectiveness ratios, expressed as cost per year of life saved (YLS). Assuming lifelong efficacy and 80% coverage, pre-adolescent HPV vaccination alone was projected to reduce the lifetime risk of cervical cancer by 55%, which was greater than any strategy of screening alone. When cost per vaccinated girl was I$10 (approximately $2 per dose) or less, HPV vaccination alone was cost saving. Pre-adolescent vaccination and HPV DNA testing five times per lifetime, starting at age 35 years, reduced the lifetime cervical cancer risk by 70%, and had a cost-effectiveness ratio less than Thailand's GDP per capita (I$8100), provided the cost per vaccinated girl was I$200 or less. Low cost pre-adolescent HPV vaccination followed by HPV screening five times per lifetime is an efficient strategy for Thailand. Costs may need to be lower, however, for this strategy to be affordable. If vaccination is not feasible, HPV DNA testing five times per lifetime is efficient.


Stepwise photoinhibition of photosystem II. Studies with Synechocystis species PCC 6803 mutants with a modified D-E loop of the reaction center polypeptide D1.
Several mutant strains of Synechocystis sp. PCC 6803 with large deletions in the D-E loop of the photosystem II (PSII) reaction center polypeptide D1 were subjected to high light to investigate the role of this hydrophilic loop in the photoinhibition cascade of PSII. The tolerance of PSII to photoinhibition in the autotrophic mutant DeltaR225-F239 (PD), when oxygen evolution was monitored with 2,6-dichloro-p-benzoquinone and the equal susceptibility compared with control when monitored with bicarbonate, suggested an inactivation of the QB-binding niche as the first event in the photoinhibition cascade in vivo. This step in PD was largely reversible at low light without the need for protein synthesis. Only the next event, inactivation of QA reduction, was irreversible and gave a signal for D1 polypeptide degradation. The heterotrophic deletion mutants DeltaG240-V249 and DeltaR225-V249 had severely modified QB pockets, yet exhibited high rates of 2,6-dichloro-p-benzoquinone-mediated oxygen evolution and less tolerance to photoinhibition than PD. Moreover, the protein-synthesis-dependent recovery of PSII from photoinhibition was impaired in the DeltaG240-V249 and DeltaR225-V249 mutants because of the effects of the mutations on the expression of the psbA-2 gene. No specific sequences in the D-E loop were found to be essential for high rates of D1 polypeptide degradation.


Control Analysis of Protein-Protein Interaction Network Reveals Potential Regulatory Targets for MYCN.
MYCN is an oncogenic transcription factor of the MYC family and plays an important role in the formation of tissues and organs during development before birth. Due to the difficulty in drugging MYCN directly, revealing the molecules in MYCN regulatory networks will help to identify effective therapeutic targets. We utilized network controllability theory, a recent developed powerful tool, to identify the potential drug target around MYCN based on Protein-Protein interaction network of MYCN. First, we constructed a Protein-Protein interaction network of MYCN based on public databases. Second, network control analysis was applied on network to identify driver genes and indispensable genes of the MYCN regulatory network. Finally, we developed a novel integrated approach to identify potential drug targets for regulating the function of the MYCN regulatory network. We constructed an MYCN regulatory network that has 79 genes and 129 interactions. Based on network controllability theory, we analyzed driver genes which capable to fully control the network. We found 10 indispensable genes whose alternation will significantly change the regulatory pathways of the MYCN network. We evaluated the stability and correlation analysis of these genes and found EGFR may be the potential drug target which closely associated with MYCN. Together, our findings indicate that EGFR plays an important role in the regulatory network and pathways of MYCN and therefore may represent an attractive therapeutic target for cancer treatment.


Inhibitory effects of chlorophyllin on 7,12-dimethylbenz[a]anthracene-induced bacterial mutagenesis and mouse skin carcinogenesis.
Chlorophyllin (CHL), a water-soluble derivative of chlorophyll, has been used for the treatment of several abnormal human conditions without apparent toxicity. Recent studies have revealed that CHL has the excellent chemopreventive potential. In the present investigation, we have found the inhibitory activities of CHL against 7,12-dimethylbenz[a]anthracene (DMBA)-induced mutagenesis in Salmonella typhimurium TA100 and also on DMBA-initiated and 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-promoted mouse skin tumor formation. The incidence and the multiplicity of skin tumors were not significantly decreased in mice by a single topical application of CHL prior to the DMBA treatment, but there was a marked suppression of papillomagenesis in mice treated with CHL during the promotional stage. Furthermore, the formation of DMBA-induced papillomagenesis was reduced in all mice that had received CHL for 6 weeks following treatment with TPA for 6, 18 and 24 weeks. These results indicate that CHL can inhibit both tumor promotion and the progression of papillomagenesis in the two-stage mouse skin carcinogenesis induced by DMBA and TPA.


Optimized outcome prediction in breast cancer by combining the 70-gene signature with clinical risk prediction algorithms.
Clinical guidelines for breast cancer treatment differ in their selection of patients at a high risk of recurrence who are eligible to receive adjuvant systemic treatment (AST). The 70-gene signature is a molecular tool to better guide AST decisions. The aim of this study was to evaluate whether adding the 70-gene signature to clinical risk prediction algorithms can optimize outcome prediction and consequently treatment decisions in early stage, node-negative breast cancer patients. A 70-gene signature was available for 427 patients participating in the RASTER study (cT1-3N0M0). Median follow-up was 61.6 months. Based on 5-year distant-recurrence free interval (DRFI) probabilities survival areas under the curve (AUC) were calculated and compared for risk estimations based on the six clinical risk prediction algorithms: Adjuvant! Online (AOL), Nottingham Prognostic Index (NPI), St. Gallen (2003), the Dutch National guidelines (CBO 2004 and NABON 2012), and PREDICT plus. Also, survival AUC were calculated after adding the 70-gene signature to these clinical risk estimations. Systemically untreated patients with a high clinical risk estimation but a low risk 70-gene signature had an excellent 5-year DRFI varying between 97.1 and 100 %, depending on the clinical risk prediction algorithms used in the comparison. The best risk estimation was obtained in this cohort by adding the 70-gene signature to CBO 2012 (AUC: 0.644) and PREDICT (AUC: 0.662). Clinical risk estimations by all clinical algorithms improved by adding the 70-gene signature. Patients with a low risk 70-gene signature have an excellent survival, independent of their clinical risk estimation. Adding the 70-gene signature to clinical risk prediction algorithms improves risk estimations and therefore might improve the identification of early stage node-negative breast cancer patients for whom AST has limited value. In this cohort, the PREDICT plus tool in combination with the 70-gene signature provided the best risk prediction.


Sodium butyrate improves mitochondrial function and kidney tissue injury in diabetic kidney disease via the AMPK/PGC-1α pathway.
Investigate the mechanism of how sodium butyrate (NaBut) improves mitochondrial function and kidney tissue injury in diabetic kidney disease (DKD) via the AMPK/PGC-1α pathway. Assess the effects of NaBut on glucose and insulin tolerance, urine, and gut microbial composition in db/db and db/m mice. Use flow cytometry and western blotting to detect the effects of NaBut on apoptosis, kidney mitochondrial function, and AMPK/PGC-1α signaling. Use HK-2 cells induced by high glucose (HG) to establish the DKD model in vitro and detect changes in the AMPK/PGC-1α signaling pathway and mitochondrial function after NaBut intervention. NaBut attenuated blood glucose levels and reversed increases in urine and serum levels of glucose, BUN, Ucr, TG, TC, and UAE in db/db mice. NaBut improved insulin tolerance, reversed PGC-1α and p-AMPK expression level in the kidneys of db/db mice, and improved lipid accumulation and mitochondrial function. NaBut was able to reverse the effects of elevated glucose, compound C, and siRNA-PGC on ROS and ATP levels. Additionally, it increased protein expression of PGC-1α and p-AMPK. NaBut activates the kidney mitochondrial AMPK/PGC-1α signaling pathway and improves mitochondrial dysfunction in DKD, thus protecting kidney tissue in vitro and in vivo.


Memory retrieval of smoking-related images induce greater insula activation as revealed by an fMRI-based delayed matching to sample task.
Nicotine dependence is a chronic and difficult to treat disorder. While environmental stimuli associated with smoking precipitate craving and relapse, it is unknown whether smoking cues are cognitively processed differently than neutral stimuli. To evaluate working memory differences between smoking-related and neutral stimuli, we conducted a delay-match-to-sample (DMS) task concurrently with functional magnetic resonance imaging (fMRI) in nicotine-dependent participants. The DMS task evaluates brain activation during the encoding, maintenance and retrieval phases of working memory. Smoking images induced significantly more subjective craving, and greater midline cortical activation during encoding in comparison to neutral stimuli that were similar in content yet lacked a smoking component. The insula, which is involved in maintaining nicotine dependence, was active during the successful retrieval of previously viewed smoking versus neutral images. In contrast, neutral images required more prefrontal cortex-mediated active maintenance during the maintenance period. These findings indicate that distinct brain regions are involved in the different phases of working memory for smoking-related versus neutral images. Importantly, the results implicate the insula in the retrieval of smoking-related stimuli, which is relevant given the insula's emerging role in addiction.


[Microscopic observation of the enamel microstructures of SD rats with different degrees of fluorosis].
Objective: To establish a dental fluorosis model of SD rats with various degrees, to observe the microstructures of enamel samples under scanning electron microscope and to clarify the changes of enamel microstructures with various degrees of dental fluorosis, so as to provide clinical reference for the treatment of patients with moderate and severe dental fluorosis. Methods: Thirty male SD rats (6 weeks of age) were randomly divided into 3 groups with 10 rats in each group. The control group was fed with deionized water without fluoride, the low fluoride group was fed with 50 mg/L NaF deionized water and the high fluoride group was fed with 100 mg/L NaF deionized water in order to establish the dental fluorosis model of rats. After feeding for 6 weeks, the rats were sacrificed and the mandibular incisor teeth were collected and recorded. The surface and sagittal plane of each tooth were observed by scanning electron microscopy and the enamel thickness was measured. Results: In the control group, the enamel color was brown yellow. Enamel color discoloration occurred both in low-fluoride group and high-fluoride group. The enamel color in low-fluoride group was mostly yellow and white striped while in high-fluoride group was mostly chalky white. Under electronic microscope, the enamel rods were alternately arranged and their structure was clear and plump in the control group. The enamel rods of moderate fluorosis were arranged in a straight orientation like tips of bamboo shoots. The enamel rods of severe fluorosis, however, became thinner and the tips of rods were broken. In the control group, sagittal images of enamel turned out to be a dense outer structure with clear boundaries among the inner. The structure of the middle layer was reticulated showing a clear boundary with middle and outer layers. The structure of enamel rods in the inner layers was arranged vertically and horizontally. In the moderate fluorosis group, the outer layer of the enamel became thinner and the middle layer disappeared although the boundary between the outer and middle layers was still clear. In the inner layer, the vertically arranged enamel rods seemed still clear, however the horizontal enamel rods disappeared. In the severe fluorosis group, the outer layer could not be traced. The middle layer was exposed to the air and the inner enamel rods contracted. The inner layers of the enamel had gradually become thinner with the development of the dental fluorosis. The thicknesses of inner layers in control, moderate and the severe groups were (180.71±7.01), (157.10±11.04) and (121.10±12.56) μm respectively. As for the thicknesses of the full layers in the above mentioned three groups, the same trend was observed. The thicknesses, in order of the severity of dental fluorosis, were (241.54±7.76), (207.42±14.36) and (143.79±14.60) μm. Conclusions: With the development of dental fluorosis, the outer enamel layers became thinner or disappeared and the inner enamel layers became thinner or lost its normal structure as well. It is highly recommended that the resin penetration could be used for the proper treatment of moderate and severe dental fluorosis and the strong bleaching and the micro-grinding should be used cautiously.


Perspectives of 360-Degree Cinematic Virtual Reality: Interview Study Among Health Care Professionals.
The global market for medical education is projected to increase exponentially over the next 5 years. A mode of delivery expected to drive the growth of this market is virtual reality (VR). VR simulates real-world objects, events, locations, and interactions in 3D multimedia sensory environments. It has been used successfully in medical education for surgical training, learning anatomy, and advancing drug discovery. New VR research has been used to simulate role-playing and clinical encounters; however, most of this research has been conducted with health professions students and not current health care professionals. Thus, more research is needed to explore how health care professionals experience VR with role-playing and clinical encounters. The aim of this study was to explore health care professionals' experiences with a cinematic VR (cine-VR) training program focused on role-playing and clinical encounters addressing social determinants of health, Appalachian culture, and diabetes. Cine-VR leverages 360-degree video with the narrative storytelling of cinema to create an engaging educational experience. We conducted in-depth telephone interviews with health care professionals who participated in the cine-VR training. The interviews were audio recorded and transcribed verbatim. A multidisciplinary team coded and analyzed the data using content and thematic analyses with NVivo software. We conducted 24 in-depth interviews with health care professionals (age=45.3, SD 11.3, years; n=16, 67%, women; n=22, 92%, White; and n=4, 17%, physicians) to explore their experiences with the cine-VR training. Qualitative analysis revealed five themes: immersed in the virtual world: seeing a 360-degree sphere allowed participants to immerse themselves in the virtual world; facilitated knowledge acquisition: all the participants accurately recalled the culture of Appalachia and listed the social determinants of health presented in the training; empathized with multiple perspectives: the cine-VR provided a glimpse into the real life of the main character, and participants described thinking about, feeling, and empathizing with the character's frustrations and disappointments; perceived ease of use of cine-VR: 96% (23/24) of the participants described the cine-VR as easy to use, and they liked the 360-degree movement, image resolution, and sound quality but noted limitations with the buttons on the headsets and risk for motion sickness; and perceived utility of cine-VR as a teaching tool: participants described cine-VR as an effective teaching tool because it activated visual and affective learning for them. Participants emphasized the realism of the cine-VR training program. They attributed the utility of the cine-VR to visual learning in conjunction with the emotional connection to the VR characters. Furthermore, participants reported that the cine-VR increased their empathy for people. More research is needed to confirm an association between the level of immersion and empathy in cine-VR training for health care professionals.


Prospective effects of social support on internalized homonegativity and sexual identity concealment among middle-aged and older gay men: a longitudinal cohort study.
Middle-aged and older gay men experience higher rates of depression and anxiety compared to their heterosexual counterparts, with internalized homonegativity and sexual identity concealment known to be major stress-related contributors. This study examined the prospective effect of different types and sources of social support on internalized homonegativity and sexual identity concealment experienced among middle-aged and older gay men. A longitudinal survey involving two waves of data collection separated by 12 months was conducted among a cohort of 186 gay-identified men aged 40 years and older. Two types of social support were found to be important. Greater baseline tangible or practical support independently predicted lower internalized homonegativity at 12-month follow-up, while greater baseline emotional or psychological support independently predicted a lower tendency toward sexual identity concealment at 12-month follow-up. Greater baseline support from community or government agencies, such as health services and support organizations, predicted higher internalized homonegativity at 12-month follow-up. These findings suggest that tangible and emotional support may be beneficial in reducing internalized homonegativity and sexual identity concealment among middle-aged and older gay men. Ensuring that services provide environments that do not compound the stressful impact of stigma also appears to be important.


The minimal invasive surgery anterior approach with supine patient positioning: a step-wise introduction of technique.
The purpose of the review is to report the modified Smith-Peterson technique for use with minimal invasive total hip arthroplasty (MIS THA) with supine patient positioning. The modified Smith-Peterson technique allows for direct visualisation of the acetabulum and is an internerval approach. The MIS anterior approach with supine patient positioning offers a complete intermuscular and internervous access to the hip joint. The use of this technique yields a reduction of muscular trauma, intraoperative bloodloss, and postoperative rehabilitation. The reduction of soft tissue trauma may lead to minimal postoperative pain. While cosmesis is never a reason for using MIS, it does yield great patient satisfaction. The MIS anterior approach with supine patient positioning is a safe and reproducible method, offering an alternative option in MIS THA.


Calibration-free extraction of constitutive parameters of magnetically coupled anisotropic metamaterials using waveguide measurements.
A calibration-free method for extraction of electromagnetic properties of magnetically coupled anisotropic biaxial metamaterial (MM) slabs from waveguide measurements is proposed. It relies on three measurement steps (thru, empty line, and the same line arbitrarily loaded by the MM slab) to extract electromagnetic properties. It is evaluated against another calibration-dependent method for retrieval of electromagnetic properties of a MM slab constructed by C-shaped resonators. From evaluation analysis, we note that our method, as compared to the calibration-dependent method, not only accurately extracts electromagnetic properties without requiring the use of expensive calibration standards but also is reference-invariant.


Algal pheromone biosynthesis: stereochemical analysis and mechanistic implications in gametes of Ectocarpus siliculosus.
During sexual reproduction, female gametes or eggs of brown algae release pheromones to attract their male mating partners. The biologically active compounds comprise linear or alicyclic unsaturated hydrocarbons derived from the aliphatic terminus of C(20) polyunsaturated fatty acids (PUFAs) by oxidative cleavage. The current study addresses the stereochemical course of the pheromone biosynthesis using female gametes of the marine brown alga E. siliculosus and chiral deuterium-labeled arachidonic acids. The biosynthetic sequence is likely to proceed via an intermediary 9-hydroperoxyarachidonic acid, which is cleaved with loss of the C(16)-H(R) into the C(11)-hydrocarbon dictyopterene C and 9-oxonona-(5Z,7E)-dienoic acid.


Observation of the inverse Doppler effect.
We report experimental observation of an inverse Doppler shift, in which the frequency of a wave is increased on reflection from a receding boundary. This counterintuitive effect has been produced by reflecting a wave from a moving discontinuity in an electrical transmission line. Doppler shifts produced by this system can be varied in a reproducible manner by electronic control of the transmission line and are typically five orders of magnitude greater than those produced by solid objects with kinematic velocities. Potential applications include the development of tunable and multifrequency radiation sources.


Rat ultrasonic vocalizations and novelty-induced social and non-social investigation behavior in a seminatural environment.
Although rats are known to emit ultrasonic vocalizations (USVs), it remains unclear whether these calls serve an auditory communication purpose. For USVs to be part of communication, the vocal signals will need to be a transfer of information between two or more conspecifics, and with the possibility to induce changes in the behavior of the recipient. Therefore, the aim of our study was to investigate the role of USVs in adult rats' social and non-social investigation strategies when introduced into a large novel environment with unfamiliar conspecifics. We quantified a wide range of social and non-social behaviors in the seminatural environment, which could be affected by subtle signals, including USVs. We found that during the first hour in the seminatural environment the ability to vocalize did not affect how quickly adult rats met each other, their overall social investigation behavior, their passive social behavior nor their aggressive behavior. Furthermore, the non-social exploratory behaviors and behaviors reflecting anxiety/stress-like states were also unaffected. These results demonstrated that a disability to vocalize did not result in significant disadvantages (or changes) compared to intact conspecifics regarding social and non-social behaviors. This suggests that other (multi)sensory cues are more relevant in social interactions than USVs.


Role of proton ordering in adsorption preference of polar molecule on ice surface.
Adsorption of polar monomers on ice surface, relevant to the physical/chemical reaction in ice clouds as well as growth of ice, remains an open issue partially due to the unusual surface characteristics with protons at the top layer of ice. Using first-principle calculations, we explore the adsorption properties of ice surface in terms of a surface proton order parameter, which characterizes the inhomogeneity of the dangling atoms on ice surface. We show that, due to an effective electric field created by dangling OH bonds and lone pairs of water molecules not only directly neighboring but also further away from the adsorbed polar molecule on the ice surface, the adsorption energy of polar monomer on ice surface exhibits large variance and a strong correlation with the proton order parameter of ice surface. Our results about the positive correlation between the inhomogeneity of ice surface and adsorption energies suggest that the physical/chemical reactions as well as the growth of ice may prefer to occur firstly on surfaces with larger proton order parameter.


Isolation of Kaeng Khoi virus from dead Chaerephon plicata bats in Cambodia.
A virus isolated from dead Chaerephon plicata bats collected near Kampot, Cambodia, was identified as a member of the family Bunyaviridae by electron microscopy. The only bunyavirus previously isolated from Chaerephon species bats in South-East Asia is Kaeng Khoi (KK) virus (genus Orthobunyavirus), detected in Thailand over 30 years earlier and implicated as a public health problem. Using RT-PCR, nucleotide sequences from the M RNA segment of several virus isolates from the Cambodian C. plicata bats were found to be almost identical and to differ from those of the prototype KK virus by only 2.6-3.2 %, despite the temporal and geographic separation of the viruses. These results identify the Cambodian bat viruses as KK virus, extend the known virus geographic range and document the first KK virus isolation in 30 years. These genetic data, together with earlier serologic data, show that KK viruses represent a distinct group within the genus Orthobunyavirus.


Evaluation of polyphenol composition and biological activities of two samples from summer and winter seasons of Ligularia fischeri var. Spiciformis Nakai.
Phenolic contents, antioxidant and antimicrobial activities were determined by two samples from summer (June) and winter (December) seasons of Ligularia fischeri var. spiciformis Nakai. A total of 24 phenolic compounds were identified by ultra-performance liquid chromatography (UPLC) analysis. Myricetin (1964.35 and 1829.12 μg/g) was the most dominant flavonol compared to quercetin and kaempferol. Salicylic acid (222.80 and 215.25 μg/g) was the most important phenolic compound compared to pyrogallol, caffeic acid, gentisic acid, o-coumaric acid, gallic acid, protocatechuic acid and ferulic acid in summer (June) and winter (December) seasons. Phenolic contents and antioxidant capacities were estimated for the various solvent extracts (petroleum ether, butanol, ethyl acetate, methanol and water). Ethyl acetate extract exhibited the highest phenolic (332.64 and 299.44 mg/g gallic acid equivalent) and flavonoid contents (5.72 and 5.29 mg/g quercetin equivalent) and also the strongest antioxidant activity in summer and winter seasons. Due to these metabolic variations, the antioxidant and antimicrobial activities were increased with summer seasons compared to winter seasons. Our study shows that the samples collected in June had higher phenolic compounds, stronger antioxidative and antimicrobial activity than the samples of L. fischeri leaf extracts collected in December.


MMG and EMG responses during 25 maximal, eccentric, isokinetic muscle actions.
The purpose of this study was to examine the patterns of the mechanomyographic (MMG) and electromyographic (EMG) amplitude responses of the vastus lateralis (VL), rectus femoris (RF), and vastus medialis (VM) during 25 maximal, eccentric, isokinetic muscle actions. Seven healthy adult females (22.1 +/- 2.4 yr) completed 25 maximal, eccentric muscle actions on a calibrated Cybex 6000 isokinetic dynamometer. EMG and MMG signals were obtained from the VL, RF, and VM during this test, and polynomial regression models were used to examine the relationships for normalized torque, EMG amplitude, and MMG amplitude versus repetition number. There was a cubic relationship (P < 0.05) between torque and repetition number. EMG amplitude demonstrated a positive linear relationship (P < 0.05) for the VL, a quadratic relationship (P < 0.05) for the RF, and no relationship (P > 0.05) for the VM over the 25 repetitions. MMG amplitude demonstrated a negative linear relationship (P < 0.05) for both the VL and VM, and a cubic relationship (P < 0.05) for the RF over the 25 repetitions. The results of the present study demonstrated differences between the VL, RF, and VM muscles for the patterns of EMG and MMG responses across the 25 maximal, eccentric muscle actions. These findings suggested that there are muscle-specific motor control strategies during repeated eccentric, isokinetic muscle actions.


Proliferation gradients in the inner ear of the monkey (Macaca mulatta) embryo.
Radioautographic studies were conducted on early developmental stages of the inner ear in monkey (Macaca mulatta) embryos ranging in age from stage 11 to stage 20 (25 to 39 days' gestation) and labeled for one hour with [3H] thymidine. The most active period of proliferation occurred at stage 13, at which time all regions of the otocyst, including the endolymphatic duct, were heavily labeled. In subsequent stages the dorsal portion of the endolymphatic duct failed to incorporate label, whereas proliferation continued in the ventral portion of the duct near its communication with the medial wall of the otocyst. The duct thus appeared to grow upward as a result of cell division from below. Mitotic activity continued in the remainder of the otocyst although there was a progressive temporal decrease in the labeling index. Spatial gradients also occurred, with dorsal and lateral regions less active than ventral and medial ones. As the otocyst differentiated, the cells in various areas became attenuated. These flattened cells were still capable of undergoing cell division, as indicated by the presence of labeled nuclei in all regions of the utricle, saccule, semicircular ducts, and cochlear duct.


In silico and in vitro metabolism studies of the new synthetic opiate AP-237 (bucinnazine) using bioinformatics tools.
The recent emergence of new synthetic opioids (NSOs) compounds in the illicit market is increasingly related to fatal cases. Identification and medical care of NSO intoxication cases are challenging, particularly due to high frequency of new products and extensive metabolism. As the study of NSO metabolism is crucial for the identification of these drugs in cases of intoxication, we aimed to investigate the metabolism of the piperazine NSO AP-237 (= bucinnazine). Two complementary approaches (in silico and in vitro) were used to identify putative AP-237 metabolites which could be used as consumption markers. In silico metabolism studies were realized by combining four open access softwares (MetaTrans, SyGMa, Glory X, Biotransformer 3.0). In vitro experiments were performed by incubating AP-237 (20 µM) in differentiated HepaRG cells during 0 h, 8 h, 24 h or 48 h. Cell supernatant were extracted and analyzed by liquid chromatography coupled to high-resolution mass spectrometry and data were reprocessed using three strategies (MetGem, GNPS or Compound Discoverer®). A total of 28 phase I and six phase II metabolites was predicted in silico. Molecular networking identified seven putative phase I metabolites (m/z 203.154, m/z 247.180, m/z 271.180, two m/z 289.191 isomers, m/z 305.186, m/z 329.222), including four previously unknown metabolites. Overall, this cross-disciplinary approach with molecular networking on data acquired in vitro and in silico prediction enabled to propose relevant candidate as AP-237 consumption markers that could be added to mass spectrometry libraries to help diagnose intoxication.


Alternative antibody for the detection of CA19-9 antigen: a European multicenter study for the evaluation of the analytical and clinical performance of the Access GI Monitor assay on the UniCel Dxl 800 Immunoassay System.
Gastrointestinal cancer antigen CA19-9 is known as a valuable marker for the management of patients with pancreatic cancer. The analytical and clinical performance of the Access GI Monitor assay (Beckman Coulter) was evaluated on the UniCel Dxl 800 Immunoassay System at five different European sites and compared with a reference method, defined as CA19-9 on the Elecsys System (Roche Diagnostics). Total imprecision (%CV) of the GI Monitor ranged between 3.4% and 7.7%, and inter-laboratory reproducibility between 3.6% and 4.0%. Linearity upon dilution showed a mean recovery of 97.4% (SD + 7.2%). Endogenous interferents had no influence on GI Monitor levels (mean recoveries: hemoglobin 103%, bilirubin 106%, triglycerides 106%). There was no high-dose hook effect up to 115,000 kU/L. Clinical performance investigated in sera from 1811 individuals showed a good correlation between the Access GI Monitor and Elecsys CA19-9 (R = 0.959, slope = 1.004, intercept = +0.17). GI Monitor serum levels were low in healthy individuals (n = 267, median = 6.0 kU/L, 95th percentile=23.1 kU/L), higher in individuals with various benign diseases (n = 550, medians = 5.8-13.4 kU/L, 95th percentiles = 30.1-195.5 kU/L) and even higher in individuals suffering from various cancers (n = 995, medians = 8.4-233.8 kU/L, 95th percentiles = 53.7-13,902 kU/L). Optimal diagnostic accuracy for cancer detection against the relevant benign control group by the GI Monitor was found for pancreatic cancer [area under the curve (AUC) 0.83]. Results for the reference CA19-9 assay were comparable (AUC 0.85). The Access GI Monitor provides very good methodological characteristics and demonstrates an excellent analytical and clinical correlation with the Elecsys CA19-9. The GI Monitor shows the best diagnostic accuracy in pancreatic cancer. Our results also suggest a clinical value of the GI Monitor in other cancers.


Blind axillar venous access.
The axillary vein has been suggested as an alternate site for venous access to avoid the "subclavian crush phenomenon. " Many techniques have been used to access this structure. They include complicated anatomical landmarks, contrast venography, Doppler, and ultrasound. A simple technique using the basic anatomical landmark of the deltopectoral groove and a blind venous stick has been used successfully in 165 of 168 consecutive pacemaker and ICD procedures; there were only three failures. These required an alternate approach. With a thorough knowledge of the regional anatomy, the axillary vein can be safely used as a primary site of venous access.


Aspects of delayed implantation in the roe deer (Capreolus capreolus).
Embryonic diapause in the roe deer is initiated at the blastocyst stage of development and persists for a period of 5 months. In common with other species exhibiting such diapause, quiescence of the roe deer blastocyst is associated with inhibition of endometrial secretory activity. The termination of diapause is characterized by the rapid elongation of the chorionic sac to a length of 30 cm and the simultaneous release of a secretion from the endometrial ducts and glands. The factors responsible for inducing this change in endometrial secretory activity are unknown. Although increases in plasma concentrations of oestrogen and progesterone are observed at the time of termination of diapause, these changes appear to be a consequence rather than a cause of embryonic growth.


[How to prescribe sport in patients?]
Physical activity is a non-pharmacological treatment validated by the Haute Autorité de santé since 2011. Since March 2017, a new French law recalls that generalist physicians can prescribe physical activity to their patients suffering from a long-term financial exonerating condition. This physical activity will be adapted to the individual physical capacities and medical risk of the patient. After assessing his/her functional abilities, the attending physician will send the patient, with his or her agreement, to a supervisory structure employing specifically trained professional for physical activity. It should be noted that neither the prescription nor the dispensation of adapted physical activity are covered by health insurance. The introduction of training in the prescription of physical activity in the framework of medical studies will be indispensable for the success of this law.


Knowledge of and attitudes toward nonpharmacological interventions for treatment of behavior symptoms associated with dementia: a comparison of physicians, psychologists, and nurse practitioners.
Behavior problems are common in nursing homes. Current guidelines recommend nonpharmacological interventions (NPHIs) as first-line treatment, but pharmacological regimens (PIs) continue to be used. Given differences in background and training of those who treat behavior problems in residents, we compared attitudes of physicians (MDs), psychologists (PhDs), and nurse practitioners (NPs) concerning PI and NPHI usage as well as knowledge of NPHIs. One hundred and eight MDs, 36 PhDs, and 89 NPs responded to a web-based questionnaire that captured level of agreement with statements concerning treatment of behavior symptoms and familiarity with NPHIs. NPs were the most favorable toward NPHIs. MDs were significantly more favorable to the use of PIs than were PhDs, with attitudes of NPs falling in between. All felt that NPHI usage should increase and that NPHIs should be implemented before using PIs but also believed that PIs work well for behavior problems. MDs had significantly lower knowledge of NPHIs than PhDs or NPs. Overall, NPHI knowledge was similar for PhDs and NPs, although they differed on their use of specific interventions. As levels of knowledge and familiarity with NPHIs differed among providers, it is conceivable that all might benefit from training and experience with a wider range of NPHIs. Future studies might evaluate the impact of a uniform understanding of NPHI on communication and teamwork in nursing homes and examine ways to enhance a multidisciplinary approach that would allow for the tailoring and individualization that is required of successful interventions.


The prognosis and treatment of dislocated total hip arthroplasties with a 22 mm head.
We studied the risk of recurrent dislocation in 121 primary and 39 revision Charnley or Charnley hybrid total hip arthroplasties which had been treated for a primary dislocation between 1979 and 1995. Only 35% of these hips had no further dislocation or a revision for instability within one year. The rates of survival gradually declined with time or if a second, third or fourth dislocation occurred. The risk of recurrence was greater in men, but was not related to age, diagnosis, time of the first dislocation or whether the index operation had been a primary or a revision procedure. Operative treatment included 15 reoperations leaving intact components, 50 revisions, and permanent removal of the femoral stem in seven patients. The operation was successful in four patients with reoperations and in 36 who had an exchange procedure within two years. Treatment was successful in 35 of 49 hips in which it was possible to correct a technical error compared with 5 out of 16 hips in which malposition of the components was not seen (p = 0.007).


An inverse association between ovarian cysts and breast cancer in the breast cancer family registry.
Ovarian cysts of several types are common in women of reproductive age. Their etiology is not well understood but is likely related to perturbations in the hypothalamic-pituitary-gonadal axis. The relationship of ovarian cysts to breast cancer risk is not known, although a negative association with polycystic ovarian syndrome has been reported. Incident, invasive female breast cancer cases, population-based controls and unaffected sisters of cases were studied from 3 countries participating in the Breast Cancer Family Registry: Melbourne and Sydney, Australia; the San Francisco Bay Area, USA; and Ontario, Canada. Using the same questionnaire, information was collected on self-reported history of ovarian cysts and other risk factors. Analyses were based on 3,049 cases, 2,344 population controls and 1,934 sister controls from all sites combined. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using both unconditional and conditional logistic regression using an offset term to account for sampling fractions at 2 of the sites. A significantly reduced risk of breast cancer was observed for women reporting a history of ovarian cysts (OR = 0.70, 95% CI 0.59-0.82, among all cases and all controls). This risk estimate was similar regardless of control group used, within all 3 sites and in both premenopausal and postmenopausal women (ORs ranging from 0.68-0.75, all 95% CI excluded 1.00). A self-reported history of ovarian cysts was strongly and consistently associated with a reduced risk of breast cancer. Further study of ovarian cysts may increase our understanding of hormonal and other mechanisms of breast cancer etiology.


Massively parallel pyrosequencing of the mitochondrial genome with the 454 methodology in forensic genetics.
of sequencing of whole mitochondrial genome, HV1 and HV2 DNA with the second generation system (SGS) Roche 454 GS Junior were compared with results of Sanger sequencing and SNP typing with SNaPshot single base extension detected with MALDI-TOF and capillary electrophoresis. We investigated the performance of the software analysis of the data, reproducibility, ability to sequence homopolymeric regions, detection of mixtures and heteroplasmy as well as the implications of the depth of coverage. We found full reproducibility between samples sequenced twice with SGS. We found close to full concordance between the mtDNA sequences of 26 samples obtained with (1) the 454 SGS method using a depth of coverage above 100 and (2) Sanger sequencing and SNP typing. The discrepancies were primarily observed in homopolymeric regions. The 454 SGS method was able to sequence 95% of the reads correctly in homopolymers up to 4 bases, and up to 6 bases could be sequenced with similar success if the results were carefully, visually inspected. The 454 technology was able to detect mixtures or heteroplasmy of approximately 10%. We detected previously unreported heteroplasmy in the GM9947A component of the NIST human mitochondrial DNA SRM-2392 standard reference material.


Aromatic hydrocarbon receptor (AhR) in the porcine theca and granulosa cells: effect of TCDD, PCB 126 and PCB 153 on the expression of AhR.
In our previous papers, we demonstrated that TCDD and PCBs (both coplanar PCB 126 and non-coplanar PCB 153) caused the decreased estradiol secretion in cultured pig ovarian follicles. However, the mechanism of action is not clearly understood. Moreover, to our knowledge, the expression of AhR in pig follicular cells was not described yet. In order to elucidate if TCDD, PCB 126, and PCB 153 can disrupt ovarian steroidogenesis via AhR-dependent mechanism, granulosa and theca cells were isolated from pig ovarian small follicles and subsequently treated with 32 pg/ml of TCDD, 100 ng/ml of PCB 153, 100 pg/ml of PCB 126 in an in vitro culture. After 3-hour exposure to the chemicals, cells were prepared for immunohistochemical analysis or collected for immunobloting, and the effects of TCDD and PCBs on the levels of AhR protein were studied. Under basal conditions, AhR staining showed a diffuse cytoplasmic pattern of distribution as well in granulosa and theca cells. However, immunocytochemical labeling of AhR in theca cells was less evident and only few cells displayed cytoplasmic staining. Immunoblotts prepared from granulosa and theca cell lysates detected a strong band of 120 kDa indicating AhR protein expression. The level of AhR expression detected by Western blot analysis was higher in granulosa cells than in theca cells. Intensive cytoplasmic and nuclear labeling corresponding to AhR protein in TCDD and PCB 126 treated granulosa cells was observed. In PCB 153 treated cells, the level of AhR staining was comparable with control. Western blot analysis showed a strong band in TCDD-treated grnulosa cells and only slightly differences compared with control in those exposed to PCB 126. The effect of both PCBs on immunocytochemical AhR staining in theca cells was less evident. Immunoblott analysis did not reveal any substantial differences between the control and TCDD- and PCBs-treated theca cells. This study has shown different AhR expression in porcine theca and granulosa cells, in respect of both its intracellular localization and cellular distribution. Both, dioxin and dioxin-like PCB activated AhRs in granulosa but not in theca cells. Differences in AhR-responsiveness of granulosa and theca cells to TCDD and dioxin-like PCB 126 are probably connected with the differences of these cells in estradiol secretion and different proliferative potential.


Genetic analysis of the pAD1 pheromone response in Streptococcus faecalis, using transposon Tn917 as an insertional mutagen.
The conjugative plasmid pAD1 (56.7 kilobases) in Streptococcus faecalis has been shown to confer a mating response to the sex pheromone cAD1 excreted by recipient strains. The response is characterized by the synthesis of a proteinaceous adhesin which coats the surface of the pAD1 -containing donor cell and facilitates the formation of mating aggregates. Donors exposed to cAD1 -containing filtrates of recipients undergo self-aggregation (clumping), an event believed to be associated with an interaction between the adhesin and a binding substance always present on the surface of both recipients and donors. To analyze the molecular processes involved in the mating response, mutants were generated by the erythromycin resistance transposon Tn917 . Transpositions to pAD1 in S. faecalis DS16 gave rise to a number of derivatives that exhibited "constitutive clumping" and the ability to transfer at high frequencies in short (10-min) matings. These mutants fell into two subclasses, which exhibited colony morphologies that were "dry" or "normal". The Tn917 insertions were mapped by restriction enzyme analysis to two separate clusters, designated traA and traB. The dry colony subclass corresponded to traA and represented a span of 1.5 kilobases, whereas the normal subclass corresponded to traB and spanned 1.3 kilobases. The two clusters were separated by 1.7 kilobases in which insertions of Tn917 did not affect the ability to respond normally to cAD1 . Neither type of constitutive clumper produced cAD1 . Another series of insertions exhibited reduced donor potential. In two cases, the reduction in transfer was three to four orders of magnitude; these mapped in traA . In two other cases, the reduction was one to two orders of magnitude. These mapped outside of traA and traB, and one was associated with an increase in plasmid copy number.


Social, cultural and economical determinants of diabetes mellitus in Kalutara district, Sri Lanka: a cross sectional descriptive study.
Sri Lanka is a country that is expected to face a high burden of diabetes mellitus (DM). There is a paucity of data on social and demographic determinants of DM, especially in the plantation sector. To describe social and economic correlates and inequalities of DM in Kalutara District. A cross sectional descriptive study was carried out among adults over the age of 35 years. A sample of 1300 individuals was selected using stratified random cluster sampling method from 65 Grama Niladari Divisions (GND), which were representative of urban, rural and plantation sectors. Twenty households were randomly selected from each division and one adult was randomly selected from each household. Data were collected using a pre-tested questionnaire. Fasting plasma blood sugar of ≥126mg/dl was used to define DM. Significance of prevalence of diseases and risk factors across different socio-economic strata were determined by chi square test for trend. Of 1234 adults who were screened (628 males), 202 (14.7%) had DM. Higher DM proportions (16.1%) were seen in the highest income quintile and in those educated up to Advanced Levels (AL) and above (17.3%). Prevalence in the urban, rural and plantation sectors were 23.6%, 15.5% and 8.5% respectively. Prevalence among Sinhalese, Tamils and Muslims were 14.4%, 29.0% and 20.0% respectively. There was a gradient in prevalence according to the unsatisfactory basic needs index of the GND with the highest proportion (20.7%) observed in the richest GND. The highest social status quintile demonstrated the highest proportion (17.4%) with diabetes mellitus. There is a higher prevalence of diabetes mellitus in the more affluent and educated segments of society. There is also a higher prevalence among urban compared to rural and estates. Sri Lanka is in an early stage of the epidemic where the wealthy people are at a higher risk of DM.


Is Geropsychology "Forever Young" on the Mother Continent? Addressing the Limited Research on Psychology of Aging in Africa.
Our paper highlights areas of interest to psychologists studying aging in Africa and what has already been learned about aspects of psychological aging from studies done in the African context. We also considered whether there are issues that are unique about aging in the African context that might challenge how aging is typically considered in psychology research. We reviewed articles recently published in major gerontology journals focusing on African older adults, with the goal of identifying key themes of current geropsychology in Africa. We identified a number of dominant themes including: subjective aging, psychosocial impact of widowhood at old age, aging in place, stress and caregiving, cognitive aging, interrelations between HIV and aging, and successful aging. The themes of psychological aging research identified as especially relevant in Africa provide a starting point for psychological aging researchers in general to engage with the African context. Many African countries may be facing common challenges, but the existing studies are not fully representative of the aging population in the continent. Creating a framework for organizing and coordinating funding will help to address the limited research on geropsychology in Africa. Collaborative research projects/programs, joint classrooms, and fellowships that support African geropsychology scholars will be helpful. Geropsychology will benefit from more engagement with the African context by applying innovative approaches such as the adoption of longitudinal designs, mixed-method research, Experience Sampling Methods, translation of measures into indigenous languages, involvement of rural samples, and implementation science and community-engaged research.


Zinc-induced formation of a coactivator complex containing the zinc-sensing transcription factor MTF-1, p300/CBP, and Sp1.
Herein, the mechanisms of transactivation of gene expression by mouse metal response element-binding transcription factor 1 (MTF-1) were investigated. Evidence obtained from coimmunoprecipitation assays revealed that exposure of the cells to zinc resulted in the rapid formation of a multiprotein complex containing MTF-1, the histone acetyltransferase p300/CBP, and the transcription factor Sp1. Down-regulation of endogenous p300 expression by small interfering RNA transfection significantly decreased zinc-dependent metallothionein I (MT-I) gene transcription without altering induction of zinc transporter 1 (ZnT1). MTF-1 independently facilitated the recruitment of Sp1 and p300 to the protein complex in response to zinc. Mutagenesis demonstrated that the acidic domain, one of three transactivation domains of MTF-1, is required for recruitment of p300 but not Sp1 as well as for zinc-dependent activation of MT-I gene transcription. Furthermore, mutation of leucine residues (L-->A) within a nuclear exclusion signal in the MTF-1 acidic domain impaired recruitment of p300 and zinc-dependent activation of the MT-I gene. Nuclear magnetic resonance characterization of an isolated protein fragment corresponding to the MTF-1 acidic region demonstrated that this region is largely unstructured in the presence and absence of excess stoichiometric amounts of zinc. This suggests that the mechanism by which MTF-1 recruits p300 to this complex involves extrinsic-zinc-dependent steps. These studies reveal a novel zinc-responsive mechanism requiring an acidic region of MTF-1 that functions as a nuclear exclusion signal as well as participating in formation of a coactivator complex essential for transactivation of MT-I gene expression.