Game of Cruxes: co-designing a game for scientists and stakeholders for identifying joint problems.
Scientists increasingly cross their disciplinary boundaries and connect with local stakeholders to jointly solve complex problems. Working with stakeholders means higher legitimacy and supports practical impact of research. Games provide a tool to achieve such transdisciplinary collaboration. In this paper, we explore the use of a game in a participatory project where scientists and local stakeholders are seeking and defining a joint problem. The literature is clear that this step is essential but remains short on concrete methods. Here, we explore this potential in practice. We conducted parallel participatory processes in two alpine regions considered as socio-ecological system (SES) in Switzerland and France, both vulnerable to global change. Based on these two case studies, we co-constructed a game, integrating scientific concerns about key land use, climate change and socio-economic elements of a mountain SES (tourism, agriculture, housing and demography). With the game, we assessed the existence of joint problems connecting scientific and local interests. The game successfully engaged participants at both sites over 11 game sessions, showing potential of use in other transdisciplinary settings. By covering a wide array of issues, the game created a discussion space for listing problems and identifying where scientist and stakeholder interests overlap. In Switzerland, the game revealed no pressing joint problem to be addressed. In France, game sessions revealed, among other problems, an enduring and complex issue regarding the co-existence of inhabitants and powerful institutions. Having demonstrated the capacity of this game for joint-problem assessment, we believe other participatory research in similar SES could benefit from an early use of such an approach to frame the potential for collaboration. The online version contains supplementary material available at 10.1007/s11625-021-00983-2.


Dysphagia associated with unilateral vocal cord immobility after cardiovascular surgery.
Dysphagia associated with unilateral vocal cord immobility (UVCI) has received much less attention than did voice and phonation. The aim of this descriptive study was to evaluate the outcome of dysphagia associated with UVCI. Between June 2006 and September 2009, 69 hospitalized patients who underwent cardiovascular surgery were referred for dysphagia. Video endoscopic evaluation of swallowing was used for the detection of swallowing difficulties. Severity of dysphagia was assessed using the Dysphagia Severity Scale. Among the 69 patients, 31 UVCI patients who underwent video endoscopic evaluation of swallowing at least twice were used for analysis. All patients had severe to mild dysphagia at the first evaluation. Nineteen patients recovered from the UVCI at a mean follow-up of 125 days, whereas 12 had persistent UVCI at a mean follow-up of 216 days. Dysphagia Severity Scale at the first evaluation was not significantly different in both groups. At the last follow-up, the Dysphagia Severity Scale improved considerably without a significant difference in the magnitude of improvement in both groups. In the recovered and persistent UVCI groups, 16 and 7 patients, respectively, resumed their regular diets. Dysphagia associated with UVCI after surgery recovers, irrespective of the functional results of the UVCI.


Estimating the economic value to societies of the impact of health research: a critical review.
Estimating the economic value to societies of health research is a complex but essential step in establishing and justifying appropriate levels of investment in research. The practical difficulties encountered include: identifying and valuing the relevant research inputs (when many pieces of research may contribute to a clinical advance); accurately ascribing the impact of the research; and appropriately valuing the attributed economic impact. In this review, relevant studies identified from the literature were grouped into four categories on the basis of the methods used to value the benefits of research. The first category consists of studies that value the direct cost savings that could arise from research leading either to new, less-costly treatments or to developments such as vaccines that reduce the number of patients needing treatment. The second category comprises studies that consider the value to the economy of a healthy workforce. According to this "human capital" approach, indirect cost savings arise when better health leads to the avoidance of lost production. The third category includes studies that examine gains to the economy in terms of product development, consequent employment and sales. The studies placed in the fourth category measure the intrinsic value to society of the health gain, by placing a monetary value on a life. The review did not identify any consistency of methodology, but the fourth approach has most promise as a measure of social value. Many of the studies reviewed come from industrialized nations and a proposal is made by the present reviewers for an international initiative, covering developed and developing countries, to undertake further methodological analysis and testing.


HIV/AIDS Indicators for Clinical Care Quality Assessment: Relevance and Utility Assessed by Health Professionals.
After conducting a systematic review of quality indicators for assessing HIV/AIDS clinical care, we aimed to assess the clinical relevance and practice utility of those indicators from the point of view of HIV/AIDS physician experts. This is an observational, cross-sectional study, in which we selected, by convenience, physicians who work in an Infectious Disease Department of a central hospital to complete two questionnaires with a core set of indicators to establish the most relevant and useful indicators for assessment of the clinical HIV/AIDS care. A Likert scale was used to rank the indicators. Eleven of thirteen physicians filled two questionnaires. From the initial list of 53 quality indicators, 21 were identified as the most relevant and useful in HIV/AIDS clinical care. The internal consistency for clinical relevance in each indicators domain was for clinical signs and symptoms (p = 0.971), for therapy (p = 0.900), for prognosis (p = 0.820) and diagnosis (p = 0.733) and for practice utility were diagnosis (p = 0.934), clinical signs (p = 0.964), laboratory examinations (p = 0.947), therapy (p = 0.583) and prognosis (p = 0.368). In the process of assessing the clinical relevance and practice utility of HIV/AIDS quality care indicators, it was found that the majority of physicians agreed that diagnosis and clinical symptoms and signs indicators domains are the most important for assessing the quality of care for HIV/AIDS patients. This instrument should be considered as a diagnostic tool, allowing hospital administrators to identify if HIV/AIDS care is properly delivered or needs improvement.


Association of Positive Airway Pressure With Cardiovascular Events and Death in Adults With Sleep Apnea: A Systematic Review and Meta-analysis.
Sleep apnea (obstructive and central) is associated with adverse cardiovascular risk factors and increased risks of cardiovascular disease. Positive airway pressure (PAP) provides symptomatic relief, whether delivered continuously (CPAP) or as adaptive servo-ventilation (ASV), but the associations with cardiovascular outcomes and death are unclear. To assess the association of PAP vs control with cardiovascular events and death in patients with sleep apnea. MEDLINE, EMBASE, and the Cochrane Library were systematically searched from inception date to March 2017 for randomized clinical trials that included reporting of major adverse cardiovascular events or deaths. Two authors independently extracted data using standardized forms. Summary relative risks (RRs), risk differences (RDs) and 95% CIs were obtained using random-effects meta-analysis. The main outcomes were a composite of acute coronary syndrome (ACS) events, stroke, or vascular death (major adverse cardiovascular events); cause-specific vascular events; and death. The analyses included data from 10 trials (9 CPAP; 1 ASV) of patients with sleep apnea (N = 7266; mean age, 60.9 [range, 51.5 to 71.1] years; 5847 [80.5%] men; mean [SD] body mass index, 30.0 [5.2]. Among 356 major adverse cardiovascular events and 613 deaths recorded, there was no significant association of PAP with major adverse cardiovascular events (RR, 0.77 [95% CI, 0.53 to 1.13]; P = .19 and RD, -0.01 [95% CI, -0.03 to 0.01]; P = .23), cardiovascular death (RR, 1.15 [95% CI, 0.88 to 1.50]; P = .30 and RD -0.00 [95% CI, -0.02 to 0.02]; P = .87), or all-cause death (RR, 1.13 [95% CI, 0.99 to 1.29]; P = .08 and RD, 0.00 [95% CI, -0.01 to 0.01]; P = .51). The same was true for ACS, stroke, and heart failure. There was no evidence of different associations for CPAP vs ASV (all P value homogeneity >.24), and meta-regressions identified no associations of PAP with outcomes for different levels of apnea severity, follow-up duration, or adherence to PAP (all P values > .13). The use of PAP, compared with no treatment or sham, was not associated with reduced risks of cardiovascular outcomes or death for patients with sleep apnea. Although there are other benefits of treatment with PAP for sleep apnea, these findings do not support treatment with PAP with a goal of prevention of these outcomes.


Amphiphilic polyanhydride films promote neural stem cell adhesion and differentiation.
Several challenges currently exist for rational design of functional tissue engineering constructs within the host, which include appropriate cellular integration, avoidance of bacterial infections, and low inflammatory stimulation. This work describes a novel class of biodegradable, amphiphilic polyanhydrides with many desirable protein-material and cell-material attributes capable of confronting these challenges. The biocompatible amphiphilic polymer films were shown to release laminin in a stable and controlled manner, promote neural cell adhesion and differentiation, and evade inflammatory responses of the immune system. Using high-throughput approaches, it was shown that polymer chemistry plays an integral role in controlling cell-film interactions, which suggests that these polyanhydrides can be tailored to achieve the desired cell adhesion and differentiation while minimizing immune recognition. These findings have important implications for development of engineered constructs to regulate differentiation and target the growth of transplanted cells in stem cell-based therapies to treat nervous system disorders.


Performance of the acetic acid test when used in field conditions as a screening test for cervical cancer.
To assess if visual inspection with acetic acid (VIA) is a useful alternative screening test for cervical cancer, when used in a resource-poor setting with an existing cytology-based screening programme. Women living in Rivas district (Nicaragua), who attended the programme, were concurrently screened with VIA and Papanicolau (PAP) smear. Screening was performed by health providers who had received training in VIA and a refresher course in cytology. Women testing positive for either of the results were referred for colposcopy and biopsy when indicated. The performance of VIA was compared with PAP smear, calculating the relative true and false positive rate (RELTPR and RELFPR) and for a high threshold on biopsy (cervical intraepithelial neoplasia grade 2 or a higher grade). We determined the trade-off between both tests by calculating the ratio of extra false positives detected through extra true positives (EFP:ETP ratio). A total of 1076 patients were screened. Nearly 33% had a positive screening test. On biopsy, 7.6% had a low-grade intraepithelial lesion, 4.5% a high-grade intraepithelial lesion (HSIL) and 0.5% invasive cancer. The RELTPR (VIA to PAP) was 1.96, the RELFPR 5.02 and the EFP:ETP ratio 8.04. VIA detected twice as much HSIL and invasive cancers as the PAP smear. Yet, for every extra diagnosis, eight extra false positives had to be examined at the referral level. The VIA spectacularly increases the number of HSIL and invasive cancers detected. The high FPR is a concern for the organization of the referral level. There is a need to establish uniform criteria on test positivity and to further improve the performance in field conditions.


Two misfolding routes for the prion protein around pH 4.5.
Using molecular dynamics simulations, we show that the prion protein (PrP) exhibits a dual behavior, with two possible transition routes, upon protonation of H187 around pH 4.5, which mimics specific conditions encountered in endosomes. Our results suggest a picture in which the protonated imidazole ring of H187 experiences an electrostatic repulsion with the nearby guanidinium group of R136, to which the system responds by pushing either H187 or R136 sidechains away from their native cavities. The regions to which H187 and R136 are linked, namely the C-terminal part of H2 and the loop connecting S1 to H1, respectively, are affected in a different manner depending on which pathway is taken. Specific in vivo or in vitro conditions, such as the presence of molecular chaperones or a particular experimental setup, may favor one transition pathway over the other, which can result in very different [Formula: see text] monomers. This has some possible connections with the observation of various fibril morphologies and the outcome of prion strains. In addition, the finding that the interaction of H187 with R136 is a weak point in mammalian PrP is supported by the absence of the [Formula: see text] residue pair in non-mammalian species that are known to be resistant to prion diseases.


Can Human Pluripotent Stem Cell-Derived Cardiomyocytes Advance Understanding of Muscular Dystrophies?
Muscular dystrophies (MDs) are clinically and molecularly a highly heterogeneous group of single-gene disorders that primarily affect striated muscles. Cardiac disease is present in several MDs where it is an important contributor to morbidity and mortality. Careful monitoring of cardiac issues is necessary but current management of cardiac involvement does not effectively protect from disease progression and cardiac failure. There is a critical need to gain new knowledge on the diverse molecular underpinnings of cardiac disease in MDs in order to guide cardiac treatment development and assist in reaching a clearer consensus on cardiac disease management in the clinic. Animal models are available for the majority of MDs and have been invaluable tools in probing disease mechanisms and in pre-clinical screens. However, there are recognized genetic, physiological, and structural differences between human and animal hearts that impact disease progression, manifestation, and response to pharmacological interventions. Therefore, there is a need to develop parallel human systems to model cardiac disease in MDs. This review discusses the current status of cardiomyocytes (CMs) derived from human induced pluripotent stem cells (hiPSC) to model cardiac disease, with a focus on Duchenne muscular dystrophy (DMD) and myotonic dystrophy (DM1). We seek to provide a balanced view of opportunities and limitations offered by this system in elucidating disease mechanisms pertinent to human cardiac physiology and as a platform for treatment development or refinement.


Cellular Selenoprotein mRNA Tethering via Antisense Interactions with Ebola and HIV-1 mRNAs May Impact Host Selenium Biochemistry.
Regulation of protein expression by non-coding RNAs typically involves effects on mRNA degradation and/or ribosomal translation. The possibility of virus-host mRNA-mRNA antisense tethering interactions (ATI) as a gain-of-function strategy, via the capture of functional RNA motifs, has not been hitherto considered. We present evidence that ATIs may be exploited by certain RNA viruses in order to tether the mRNAs of host selenoproteins, potentially exploiting the proximity of a captured host selenocysteine insertion sequence (SECIS) element to enable the expression of virally-encoded selenoprotein modules, via translation of in-frame UGA stop codons as selenocysteine. Computational analysis predicts thermodynamically stable ATIs between several widely expressed mammalian selenoprotein mRNAs (e.g., isoforms of thioredoxin reductase) and specific Ebola virus mRNAs, and HIV-1 mRNA, which we demonstrate via DNA gel shift assays. The probable functional significance of these ATIs is further supported by the observation that, in both viruses, they are located in close proximity to highly conserved in-frame UGA stop codons at the 3' end of open reading frames that encode essential viral proteins (the HIV-1 nef protein and the Ebola nucleoprotein). Significantly, in HIV/AIDS patients, an inverse correlation between serum selenium and mortality has been repeatedly documented, and clinical benefits of selenium in the context of multi-micronutrient supplementation have been demonstrated in several well-controlled clinical trials. Hence, in the light of our findings, the possibility of a similar role for selenium in Ebola pathogenesis and treatment merits serious investigation.


Evidence-based practice - an incomplete model of the relationship between theory and professional work.
Current day realities of diminishing resources, reductions in spending and organizational change within health care systems have resulted in an increased emphasis on a multidisciplinary team approach to quality patient care. The movement of nursing towards more autonomous practice combined with the current trend towards 'evidence-based practice' in health care demands increased accountability in clinical decision making. This paper focuses on one aspect of nurses' clinical decision making within the demands of evidence-based practice and cardiac surgery. In this field recent advances, combined with increasing demands on health care institutions, have promoted early extubation of post-operative cardiac patients. While this remains a medical role in many institutions, an increasing number of intensive care units now consider it as a nursing role. This paper explores the realities of nurses' clinical decision making through a discussion of current practice in the extubation of patients following cardiac surgery. In addition, it considers the implications of current practice for both nurse education and the continued development of clinical nursing practice. The findings indicate that evidence-based practice appears to be an incomplete model of the relationship between theory and professional work.


Population Diversity of Campylobacter jejuni in Poultry and Its Dynamic of Contamination in Chicken Meat.
This study aimed to analyse the diversity of the Campylobacter jejuni population in broilers and to evaluate the major source of contamination in poultry meat. Eight rearing cycles over one year provided samples from three different broiler farms processed at the same slaughterhouse. A total of 707 C. jejuni were isolated from cloacal swabs before slaughter and from the breast skin of carcasses after slaughter and after chilling. All suspected Campylobacter colonies were identified with PCR assays and C. jejuni was genotyped by sequence analysis of the flaA short variable region (SVR) and by pulsed-field gel electrophoresis (PFGE) using SmaI enzyme. Phenotypic antibiotic resistance profiles were also assayed using minimal inhibitory concentration (MIC). The flocks carried many major C. jejuni clones possibly carrying over the rearing cycles, but cross contamination between farms may happen. Many isolates were resistant to fluoroquinolones, raising an issue of high public concern. Specific Campylobacter populations could be harboured within each poultry farm, with the ability to contaminate chickens during each new cycle. Thus, although biosecurity measures are applied, with a persistent source of contamination, they cannot be efficient. The role of the environment needs further investigation to better address strategies to control Campylobacter.


The Effect of Zinc Deficiency and Supplementation on Elements in the Kidney Tissue of Ovariectomized Rats: Histopathologic Changes.
The objective of the present study is to determine the effects of zinc deficiency in and zinc supplementation to ovariectomized rats on some elements in kidney tissue. The study included 40 Sprague-Dawley type adult female rats. The experimental animals were randomized into four groups with equal numbers as follows: Group 1: Control (10). Group 2: Ovariectomized control (10). Group 3: Ovariectomized + zinc supplemented (10). Group 4: Ovariectomized + zinc deficient (10). After the animals were decapitated at the end of the experiment, element levels were determined by Atomic Emission (ICP-AES) as mg/g/wet tissue for calcium, phosphate, zinc, aluminum, copper, iron, lithium, and manganese and μg/g/wet tissue for magnesium in the kidney tissue. Additionally, the tissue samples were subjected to a histopathologic assessment. An examination of the study results showed that ovariectomy significantly reduced calcium, phosphorus, and zinc levels, while zinc supplementation to the rats following ovariectomy restored the reduced element levels to normal (0.10 ± 0.03, 0.85 ± 0.16, 0.11 ± 0.03 vs 0.19 ± 0.06, 1.86 ± 0.18, 0.52 ± 0.05). Group 4, which was both ovariectomized and fed on a zinc-deficient diet, had significantly lower aluminum, copper, and lithium values. Calcification, inflammation, and sclerotic changes in group 4, the group which was fed on a zinc-deficient diet, were greater in comparison to other groups (p < 0.05). Results of the study suggest that ovariectomy + zinc deficiency leads to calcification, inflammation, and sclerotic changes in renal tissue and significantly reduces element levels, whereas zinc supplementation after ovariectomy restores the lowered element levels to normal.


The antioxidative defense system is involved in the premature senescence in transgenic tobacco (Nicotiana tabacum NC89).
α-Farnesene is a volatile sesquiterpene synthesized by the plant mevalonate (MVA) pathway through the action of α-farnesene synthase. The α-farnesene synthase 1 (MdAFS1) gene was isolated from apple peel (var. white winter pearmain), and transformed into tobacco (Nicotiana tabacum NC89). The transgenic plants had faster stem elongation during vegetative growth and earlier flowering than wild type (WT). Our studies focused on the transgenic tobacco phenotype. The levels of chlorophyll and soluble protein decreased and a lower seed biomass and reduced net photosynthetic rate (Pn) in transgenic plants. Reactive oxygen species (ROS) such as hydrogen peroxide (H2O2) and superoxide radicals (O 2 (·-) ) had higher levels in transgenics compared to controls. Transgenic plants also had enhanced sensitivity to oxidative stress. The transcriptome of 8-week-old plants was studied to detect molecular changes. Differentially expressed unigene analysis showed that ubiquitin-mediated proteolysis, cell growth, and death unigenes were upregulated. Unigenes related to photosynthesis, antioxidant activity, and nitrogen metabolism were downregulated. Combined with the expression analysis of senescence marker genes, these results indicate that senescence started in the leaves of the transgenic plants at the vegetative growth stage. The antioxidative defense system was compromised and the accumulation of reactive oxygen species (ROS) played an important role in the premature aging of transgenic plants.


A new punctal plug insertion technique to prevent intracanalicular plug migration.
To evaluate the intracanalicular migration rate during plug insertion with a new plug insertion technique compared with a standard technique. Interventional, nonrandomized, comparative study. Forty-five patients with dry eye syndrome underwent a punctal plug insertion with the new technique, and 33 patients underwent a punctal plug insertion with a standard technique at the dry eye subspecialty outpatient clinic of the Department of Ophthalmology at Keio University. Tear function examinations and ocular surface evaluations, including the Schirmer test, tear film breakup time, fluorescein and Rose Bengal vital staining scores, were performed before punctal plug insertion. Super Flex Punctum Plugs (Eagle Vision, Memphis, Tennessee, USA; Softplug-Oasis Medical Inc, Glendora, California, USA) were implanted in all subjects with both techniques. In total, 120 procedures were carried out with the new technique and 132 procedures with the standard technique. There were no statistical differences between the two groups in tear function and ocular surface staining scores (P > .05). There was no intracanalicular plug migration with the new insertion technique, whereas there were 18 incidents of intracanalicular migration of 132 standard plug insertion procedures (13.6%). The new plug insertion technique seems to be effective in eliminating intracanalicular plug migration during the insertion procedure.


A subgroup of dorsal raphe serotonergic neurons in the cat is strongly activated during oral-buccal movements.
A subgroup of approximately 25% of dorsal raphe nucleus serotonergic neurons in cat was strongly activated in association with oral-buccal movements, such as chewing, licking, and grooming. The mean magnitude of increase in neuronal activity for these cells was approximately 100% above the spontaneous waking level. However, some of these cells were activated by as much as 200-300%. The neuronal activation frequently preceded the initiation of the movement and stopped abruptly in association with either pauses in the motor sequence or with its cessation. Most of the neurons in this subgroup were also strongly and preferentially activated by somatosensory stimuli applied to the head, neck, and face. During orientation to a strong or novel stimulus, the activity of these neurons fell silent for periods of 1-5 s. These data and results from our previous studies of medullary raphe neurons are discussed within the context of the general role of serotonin in tonic and central pattern generator-related motor activity.


Climate refugia: joint inference from fossil records, species distribution models and phylogeography.
Climate refugia, locations where taxa survive periods of regionally adverse climate, are thought to be critical for maintaining biodiversity through the glacial-interglacial climate changes of the Quaternary. A critical research need is to better integrate and reconcile the three major lines of evidence used to infer the existence of past refugia - fossil records, species distribution models and phylogeographic surveys - in order to characterize the complex spatiotemporal trajectories of species and populations in and out of refugia. Here we review the complementary strengths, limitations and new advances for these three approaches. We provide case studies to illustrate their combined application, and point the way towards new opportunities for synthesizing these disparate lines of evidence. Case studies with European beech, Qinghai spruce and Douglas-fir illustrate how the combination of these three approaches successfully resolves complex species histories not attainable from any one approach. Promising new statistical techniques can capitalize on the strengths of each method and provide a robust quantitative reconstruction of species history. Studying past refugia can help identify contemporary refugia and clarify their conservation significance, in particular by elucidating the fine-scale processes and the particular geographic locations that buffer species against rapidly changing climate.


Hmga1 is required for normal sperm development.
The Hmgi protein family of chromosomal architectural factors is extensively studied for its roles in embryogenesis and its association with benign mesenchymal tumors. Although the biochemical function of Hmga1 has been studied in vitro, to provide in vivo insight into its biological function, a targeted disruption of Hmga1 was initiated. Chimeric founder mice were derived from embryonic stem (ES) cells harboring a targeted mutation in a single Hmga1 allele. These 14 different chimeric founders produced 494 black progeny. Since none of these 494 progeny were agouti, none of them were derived from ES cells. Control injections of the wild-type ES cell lines resulted in ES cell derived agouti mice, indicating that the ES cells were totipotent. Therefore, our results indicate that one intact Hmga1 allele was not sufficient for germ-line transmission of the ES cells. Seven chimeric founder mice that were examined histologically demonstrated aberrant regions in their reproductive organs. Aberrant regions of seminiferous tubules were reduced in diameter, demonstrated vacuolated Sertoli cells, and had an absolute deficiency of sperm. While the Hmga1(+/-) ES cells were shown to contribute to the formation of the epididymides, they did not significantly contribute to the testes of chimeric founder mice. No sperm isolated from any of the Hmga1(+/-) chimeric mice were shown to arise from the ES cells, as none of them contained the targeted disruption of the Hmga1 gene. Our results suggest that both alleles of Hmga1 are required for normal sperm production in the mouse.


Differential roles for group 1 mGluR subtypes in induction and expression of chemically induced hippocampal long-term depression.
Although metabotropic glutamate receptors (mGluRs) mGluR1 and mGluR5 are often found to have similar functions, there is considerable evidence that the two receptors also serve distinct functions in neurons. In hippocampal area CA1, mGluR5 has been most strongly implicated in long-term synaptic depression (LTD), whereas mGluR1 has been thought to have little or no role. Here we show that simultaneous pharmacological blockade of mGluR1 and mGluR5 is required to block induction of LTD by the group 1 mGluR agonist, (RS)-3,5-dihydroxyphenylglycine (DHPG). Blockade of mGluR1 or mGluR5 alone has no effect on LTD induction, suggesting that activation of either receptor can fully induce LTD. Consistent with this conclusion, mGluR1 and mGluR5 both contribute to activation of extracellular signal-regulated kinase (ERK), which has previously been shown to be required for LTD induction. In contrast, selective blockade of mGluR1, but not mGluR5, reduces the expression of LTD and the associated decreases in AMPA surface expression. LTD is also reduced in mGluR1 knockout mice confirming the involvement of mGluR1. This shows a novel role for mGluR1 in long-term synaptic plasticity in CA1 pyramidal neurons. In contrast to DHPG-induced LTD, synaptically induced LTD with paired-pulse low-frequency stimulation persists in the pharmacological blockade of group 1 mGluRs and in mGluR1 or mGluR5 knockout mice. This suggests different receptors and/or upstream mechanisms for chemically and synaptically induced LTD.


LoxP-FRT Trap (LOFT): a simple and flexible system for conventional and reversible gene targeting.
Conditional gene knockout (cKO) mediated by the Cre/LoxP system is indispensable for exploring gene functions in mice. However, a major limitation of this method is that gene KO is not reversible. A number of methods have been developed to overcome this, but each method has its own limitations. We describe a simple method we have named LOFT [LoxP-flippase (FLP) recognition target (FRT) Trap], which is capable of reversible cKO and free of the limitations associated with existing techniques. This method involves two alleles of a target gene: a standard floxed allele, and a multi-functional allele bearing an FRT-flanked gene-trap cassette, which inactivates the target gene while reporting its expression with green fluorescent protein (GFP); the trapped allele is thus a null and GFP reporter by default, but is convertible into a wild-type allele. The floxed and trapped alleles can typically be generated using a single construct bearing a gene-trap cassette doubly flanked by LoxP and FRT sites, and can be used independently to achieve conditional and constitutive gene KO, respectively. More importantly, in mice bearing both alleles and also expressing the Cre and FLP recombinases, sequential function of the two enzymes should lead to deletion of the target gene, followed by restoration of its expression, thus achieving reversible cKO. LOFT should be generally applicable to mouse genes, including the growing numbers of genes already floxed; in the latter case, only the trapped alleles need to be generated to confer reversibility to the pre-existing cKO models. LOFT has other applications, including the creation and reversal of hypomorphic mutations. In this study we proved the principle of LOFT in the context of T-cell development, at a hypomorphic allele of Baf57/Smarce1 encoding a subunit of the chromatin-remodeling Brg/Brahma-associated factor (BAF) complex. Interestingly, the FLP used in the current work caused efficient reversal in peripheral T cells but not thymocytes, which is advantageous for studying developmental epigenetic programming of T-cell functions, a fundamental issue in immunology. LOFT combines well-established basic genetic methods into a simple and reliable method for reversible gene targeting, with the flexibility of achieving traditional constitutive and conditional KO.


Diarrhea-predominant irritable bowel syndrome is associated with diverticular disease: a population-based study.
A subset of patients with colonic diverticular disease have chronic gastrointestinal symptoms, and some have a clinical diagnosis of irritable bowel syndrome (IBS), but whether IBS and diverticular disease are linked is uncertain. We aimed to evaluate this association in the community. A population-based, cross-sectional survey was conducted by mailing a valid symptom questionnaire to the eligible residents of Olmsted County, MN, aged 30-95 years. Colonic diverticular disease (diverticulosis and diverticulitis) was ascertained through a review of the complete medical history of all responders. Subjects with at least one relevant test (colonoscopy, computed tomography (CT) scan, CT colonography, or barium enema) were included. IBS was defined using Rome II criteria. Among 2,267 eligible respondents, there were 1,712 subjects who had undergone colon testing (76%): 919 women (54%); mean (+/-s.d.) age 65 (+/-11 years). Colonic diverticular disease was identified in 44.4% (95% confidence interval (CI) 42.1-46.8) of the subject. IBS was reported by 8.8% (95% CI 6.9-11.0) of men and 17.0% (95% CI 14.6-19.6) of women. After adjusting for age and gender, the presence of IBS was associated with an increased odds for diverticulosis (odds ratio (OR) =1.8, 95% CI 1.3-2.4) but not diverticulitis (OR=1.7, 95% CI 0.9-3.2). In those 65 years of age or older, the presence of IBS was associated with a ninefold higher odds for diverticulosis (OR=9.4, 95% CI 5.8-15.1). Relative to the non-IBS subgroup, diarrhea-predominant IBS and mixed IBS were significantly associated with an increased odds for diverticular disease (OR=1.9, 95% CI 1.1-3.2; OR=2.6, 95% CI 1.0-6.4, respectively). There is a significantly increased odds for colonic diverticulosis in subjects with IBS (relative to those without IBS). These results suggest that IBS and colonic diverticular disease may be connected.


Structural MRI findings in long-term cannabis users: what do we know?
In animal studies, tetrahydrocannabinol (THC) has been found to affect brain morphology, particularly within areas rich in cannabinoid receptors (e.g., hippocampus, cerebral cortex). While cannabis remains the most widely used illicit drug worldwide, there has been limited work investigating its effects on human brain tissue. In this paper, we conducted a systematic review of existing structural magnetic resonance imaging studies to examine whether cannabis use is associated with significant changes in brain anatomy. We identified only 13 structural neuroimaging studies, which were diverse in terms of sample characteristics (e.g., age of participants, duration and frequency of use) and methodology (e.g., image analysis). No study found global structural changes in cannabis users, although six studies reported regional alterations. While changes in the hippocampus and parahippocampus were frequently identified, the findings were inconsistent across studies. The available literature also provides some evidence that regional structural changes are associated with cannabis use patterns (particularly cumulative dosage and frequency of use), as well as measures of psychopathology (e.g., measures of depressive and psychotic symptoms). Together, these structural imaging findings suggest that THC exposure does affect brain morphology, especially in medial-temporal regions. Given the small literature available and the limitations of studies to date, further research is clearly required, particularly given the prevalence of cannabis use worldwide.


Two-stage tuberculin skin testing in individuals with human immunodeficiency virus infection. Community Programs for Clinical Research on AIDS.
In this study we estimated occurrence of the booster effect in a population infected with the human immunodeficiency virus (HIV) and assessed the relation between the booster effect, T-lymphocyte CD4 cell counts, tuberculosis risk categories, and HIV exposure categories. Patients were recruited from 13 participating sites of the Terry Beirn Community Programs for Clinical Research on AIDS (CPCRA). A two-stage tuberculin skin test was applied to 709 HIV-infected patients using the Mantoux method. An induration reading < 5 mm on the first test and > or = 5 on the second skin test defined the booster effect. Overall, 18 patients, or 2.7% (95% confidence interval, 1.6 to 4.2) experienced the booster effect. Boosted responses were seen in eight (2.1%) anergic patients, six (4.5%) nonanergic patients, and four (2.5%) with anergy status unknown. Boosting was noted in 1 of the 131 women enrolled. Age, race, CD4 cell count, injection drug use, anergy status, tuberculosis risk categories, and HIV exposure categories were not predictive of boosting. The booster effect occurs in a small percentage of HIV-infected patients tested, thus identifying small numbers of patients with latent tuberculosis infection. The two-stage procedure is probably of limited value in the diagnosis of latent tuberculosis in HIV-infected persons.


Twenty-four-hour preservation of the pig heart by a portable hypothermic perfusion system.
A portable hypothermic perfusion system for storage of hearts has been developed. The system uses the airlift pump principle, whereby the flow of gas maintains circulation of the perfusate through the heart; no other energy source is required. Performance on ex vivo functional testing of 10 pig hearts stored for 20 to 24 hr using this system (group 3) was compared with that of freshly excised hearts (group 1) and hearts stored simply in the perfusate under hypothermic conditions, but not perfused (group 2). Group 2 hearts performed less well on functional testing than those of groups 1 and 3 which showed little statistical difference, suggesting good preservation by hypothermic perfusion. This has been confirmed by orthotopic transplantation of similarly preserved baboon hearts with survival until rejection at a mean of 27 days. The importance of the various constituents of the perfusate and the significance of weight gain during the storage and reperfusion periods are discussed.


Adenosine and deoxyadenosine induces apoptosis in oestrogen receptor-positive and -negative human breast cancer cells via the intrinsic pathway.
In this study we have examined the cytotoxic effects of different concentrations of adenosine (Ado) and deoxyadenosine (dAdo) on human breast cancer cell lines. Ado and dAdo alone had little effect on cell cytotoxicity. However, in the presence of adenosine deaminase (ADA) inhibitor, EHNA, adenosine and deoxyadenosine led to significant growth inhibition of cells of the lines tested. Ado/EHNA and dAdo/EHNA-induced cell death was significantly inhibited by NBTI, an inhibitor of nucleoside transport, and 5'-amino-5'-deoxyadenosine, an inhibitor of adenosine kinase, but the effects were not affected by 8-phenyltheophylline, a broad inhibitor of adenosine receptors. The Ado/EHNA combination brought about morphological changes consistent with apoptosis. Caspase-9 activation was observed in MCF-7 and MDA-MB468 human breast cancer cell lines on treatment with Ado/EHNA or dAdo/EHNA, but, as expected, caspase-3 activation was only observed in MDA-MB468 cells. The results of the study, thus, suggest that extracellular adenosine and deoxyadenosine induce apoptosis in both oestrogen receptor-positive (MCF-7) and also oestrogen receptor-negative (MDA-MB468) human breast cancer cells by its uptake into the cells and conversion to AMP (dAMP) followed by activation of nucleoside kinase, and finally by the activation of the mitochondrial/intrinsic apoptotic pathway.


VEGF-C-dependent stimulation of lymphatic function ameliorates experimental inflammatory bowel disease.
Crohn's disease (CD) and ulcerative colitis (UC) are chronic inflammatory bowel diseases (IBDs) of unknown etiology that are associated with an aberrant mucosal immune response. Neoangiogenesis and vascular injury are observed in IBD along with increased lymphangiogenesis. While the pathogenic role of angiogenesis in IBD is well characterized, it is not clear how or if increased lymphangiogenesis promotes disease. Here, we determined that enhancing lymphangiogenesis and lymphatic function reduces experimental IBD. Specifically, we demonstrated that adenoviral induction of prolymphangiogenic factor VEGF-C provides marked protection against the development of acute and chronic colitis in 2 different animal models. VEGF-C-dependent protection was observed in combination with increased inflammatory cell mobilization and bacterial antigen clearance from the inflamed colon to the draining lymph nodes. Moreover, we found that the VEGF-C/VEGFR3 pathway regulates macrophage (MΦ) plasticity and activation both in cultured MΦs and in vivo, imparting a hybrid M1-M2 phenotype. The protective function of VEGF-C was meditated by the so-called resolving MΦs during chronic experimental colitis in a STAT6-dependent manner. Together, these findings shed light on the contribution of lymphatics to the pathogenesis of gut inflammation and suggest that correction of defective lymphatic function with VEGF-C has potential as a therapeutic strategy for IBD.


Transport characteristics of isolated newborn rabbit ileum.
Segments of ileum from newborn rabbits were mounted as flat sheets in Lucite chambers and transmural fluxes of Na, Cl, and alanine were measured in the absence of electrochemical potential gradients. In the presence of 140 mM Na, the ileum of the newborn exhibited a serosa-positive electrical PD and a corresponding short-circuit current (Isc) which was markedly enhanced by the addition of D-glucose of L-alanine to the mucosal solution. Alanine-induced increments in Isc were a saturable function of the mucosal alanine concentration. In the presence of Na, alanine was actively absorbed, and the net alanine flux was a saturable function of alanine concentration. When the Na in the bathing solutions was completely replaced by choline, Isc declined to near zero, and the response of Isc to mucosal alanine was abolished. In addition, active amino acid transport was abolished in the absence of Na. In the absence of alanine, the isolated ileum of the newborn actively absorbed Na and Cl, and the algebraic sum of the net movements of these ions accounted for Isc. In the presence of alanine, active Cl transport was abolished and Isc was equal to the net Na absorption. These results indicate that the ileum of the newborn rabbit is similar to that of the adult in its ability to actively absorb Na, Cl, and alanine, but differs from the ileum of the adult by having a greater passive permeability to ions and amino acid, and differs particularly with regard to the effect of mucosal alanine on transmural ion transport.


Islet beta-cell-specific MafA transcription requires the 5'-flanking conserved region 3 control domain.
MafA is a key transcriptional activator of islet beta cells, and its exclusive expression within beta cells of the developing and adult pancreas is distinct among pancreatic regulators. Region 3 (base pairs -8118 to -7750 relative to the transcription start site), one of six conserved 5' cis domains of the MafA promoter, is capable of directing beta-cell-line-selective expression. Transgenic reporters of region 3 alone (R3), sequences spanning regions 1 to 6 (R1-6; base pairs -10428 to +230), and R1-6 lacking R3 (R1-6(DeltaR3)) were generated. Only the R1-6 transgene was active in MafA(+) insulin(+) cells during development and in adult cells. R1-6 also mediated glucose-induced MafA expression. Conversely, pancreatic expression was not observed with the R3 or R1-6(DeltaR3) line, although much of the nonpancreatic expression pattern was shared between the R1-6 and R1-6(DeltaR3) lines. Further support for the importance of R3 was also shown, as the islet regulators Nkx6.1 and Pax6, but not NeuroD1, activated MafA in gel shift, chromatin immunoprecipitation (ChIP), and transfection assays and in vivo mouse knockout models. Lastly, ChIP demonstrated that Pax6 and Pdx-1 also bound to R1 and R6, potentially functioning in pancreatic and nonpancreatic expression. These data highlight the nature of the cis- and trans-acting factors controlling the beta-cell-specific expression of MafA.


Absence of toxicity and genotoxicity in an extract of Rubus coriifolius.
Rubus coriifolius Focke is a wild plant from the Rosaceae family. It grows in both Guatemala and Mexico. The polar extract of the aerial parts of this plant has antibacterial, anti-inflammatory, and anti-protozoal activities. These properties may explain the traditional use of this plant. In vivo and in vitro assays were used to assess the genotoxic and toxic effects of an ethanol extract of the aerial parts of R. coriifolius. Three groups of rats were orally administered the R. coriifolius extract diluted in ethanol (5%) at doses of 1.89 mg/kg body weight (low dose), 4.72 mg/kg body weight (medium dose), and 9.44 mg/kg body weight (high dose) for 3 weeks. Genotoxic/cytotoxic effects induced by the R. coriifolius ethanol extract were evaluated in vivo by a micronuclei (MN) test in rat's bone marrow cells and in vitro by MN and sister chromatid exchange (SCE) in human lymphocyte cultures. In vivo genotoxicity analyses revealed that the average number of micronucleated polychromatic erythrocytes and the polychromatic erythrocyte/red blood cell ratio at all doses were not significantly different from those of the negative control. In vitro genotoxicity analyses showed that MN, SCE, and proliferative index frequencies in a human lymphocyte cell culture were not significantly different from those of the negative control. These results demonstrate that the ethanol extract of R. coriifolius aerial parts is not toxic or mutagenic (in vitro and in vivo) and does not affect cell proliferation at the concentrations analyzed.


Isoenzymes of human lice: pediculus humanus and P. capitis.
Human lice (Phthiraptera: Pediculidae) from Africa, America and Europe were electrophoresed for 28 enzymes, with special interest in metabolic factors likely to be involved with insecticide resistance. Zymogram profiles of the body louse (Pediculus humanus L. from France and U.S.A.) and the head louse (P. capitis DeGeer from France, Madagascar, Mali & Senegal) were compared. Only esterase two enzymes, phosphoglucomutase (Pgm) and 3 (Est-3), showed electrophoretic variation. In our starch gel electrophoresis conditions, P. humanus showed three electromorphs of Pgm migrating anodally 6, 11 and 16 mm (designated alleles a, b, c, respectively). Of the putative Pgm alleles, b and c occurred in all samples of both species of lice, whereas allele a was found only in P. humanus lab strain from U.S.A. Esterase 3 had four electromorphs migrating 23, 26, 30 and 35 mm (designated alleles a, b, c and d). Among putative Est alleles, a was found only in P. capitis from Bamako (all 14 specimens aa homozygotes), allele d was found only in P. capitis from Dakar (39% frequency), whereas Est-3 alleles b and c showed apparently balanced polymorphism in all samples of both P. humanus and P. capitis except that from Bamako. Despite the limited amount of isoenzyme variation detected (only 2/31 polymorphic loci), divergences of Est-3 and Pgm among Pediculus populations may be relevant to their biosystematics and resistance.


Periprosthetic supracondylar femur fractures following total knee arthroplasty.
Periprosthetic supracondylar femur fractures following total knee arthroplasty (TKA) are an infrequent, but devastating, complication. From 1998 to 2000, we treated 30 supracondylar femur fractures above TKAs. Eighteen fractures were managed with retrograde intramedullary rod fixation (FIMR) and the other 12 fractures with traditional open reduction with internal fixation (ORIF). Follow-up averaged 3 years, with Knee Society knee scores being 84 and 82, respectively. Complications included 1 above-knee amputation for deep sepsis and 1 nonunion with varus alignment. FIMR appears to be the treatment of choice when it is feasible. However, traditional ORIF also may yield satisfactory results in those designs that cannot accommodate retrograde FIMR fixation.


An examination of ethical issues raised in the pretreatment of normal volunteer granulocyte donors with granulocyte colony-stimulating factor.
To explore some of the ethical issues surrounding the administration of granulocyte colony-stimulating factor (G-CSF) to healthy individuals for the purpose of retrieval of granulocytes. Review of the historical precedent of drug administration to normal blood donors and review of the literature concerning the side effects of G-CSF administration to healthy individuals, particularly as related to granulocyte collection. We identify and discuss some of the ethical questions regarding this issue. Although the short-term side effects of G-CSF use in normal donors are generally felt to be benign, little is known about the long-term side effects. Ethical questions regarding the administration of this drug to normal donors for the purpose of collecting large numbers of granulocytes include the following: Does the potential benefit to a patient/recipient justify the unknown risks to the medicated granulocyte donor? Who should act as an advocate for donors so that their best interests are protected? What is the role and quality of informed consent for donors undergoing G-CSF administration? Is monetary compensation appropriate for donors administered G-CSF as part of a research protocol? We recommend the establishment of a donor registry to collect the needed data on the side effects of G-CSF on normal donors. Until adequate data are collected, the use of G-CSF and similar agents in normal donors should be regarded as experimental and subject to review by institutional review boards.


Functional definition and characterization of acute traumatic coagulopathy.
To identify an appropriate diagnostic tool for the early diagnosis of acute traumatic coagulopathy and validate this modality through prediction of transfusion requirements in trauma hemorrhage. Prospective observational cohort study. Level 1 trauma center. Adult trauma patients who met the local criteria for full trauma team activation. Exclusion criteria included emergency department arrival >2 hrs after injury, >2000 mL of intravenous fluid before emergency department arrival, or transfer from another hospital. None. Blood was collected on arrival in the emergency department and analyzed with laboratory prothrombin time, point-of-care prothrombin time, and rotational thromboelastometry. Prothrombin time ratio was calculated and acute traumatic coagulopathy defined as laboratory prothrombin time ratio >1.2. Transfusion requirements were recorded for the first 12 hrs following admission. Three hundred patients were included in the study. Laboratory prothrombin time results were available at a median of 78 (62-103) mins. Point-of-care prothrombin time ratio had reduced agreement with laboratory prothrombin time ratio in patients with acute traumatic coagulopathy, with 29% false-negative results. In acute traumatic coagulopathy, the rotational thromboelastometry clot amplitude at 5 mins was diminished by 42%, and this persisted throughout clot maturation. Rotational thromboelastometry clotting time was not significantly prolonged. Clot amplitude at a 5-min threshold of ≤35 mm had a detection rate of 77% for acute traumatic coagulopathy with a false-positive rate of 13%. Patients with clot amplitude at 5 mins ≤35 mm were more likely to receive red cell (46% vs. 17%, p < .001) and plasma (37% vs. 11%, p < .001) transfusions. The clot amplitude at 5 mins could identify patients who would require massive transfusion (detection rate of 71%, vs. 43% for prothrombin time ratio >1.2, p < .001). In trauma hemorrhage, prothrombin time ratio is not rapidly available from the laboratory and point-of-care devices can be inaccurate. Acute traumatic coagulopathy is functionally characterized by a reduction in clot strength. With a threshold of clot amplitude at 5 mins of ≤35 mm, rotational thromboelastometry can identify acute traumatic coagulopathy at 5 mins and predict the need for massive transfusion.


Postnatal construction of neural circuitry in the mouse olfactory bulb.
We have undertaken a quantitative analysis of the mouse olfactory bulb to address several major questions concerning the development of neural circuitry in the postnatal mammalian brain. These are: (1) To what degree are new elements and circuits added during maturation? (2) How long do such processes go on? and (3) Does postnatal development involve a net addition of circuits and their constituent elements, or is there elimination of some portion of an initial surfeit? Using male mice of known age, weight, and length, we measured the overall size of the bulb, the numbers of processing units (glomeruli) within the bulb, the extent and complexity of postsynaptic dendrites within the glomeruli, and the number of synapses in different regions of the bulb. Between birth and the time mice reach sexual maturity at 6-7 weeks of age, the bulb increases in size by a factor of 8, the number of glomeruli by a factor of 4-5, the length of mitral cell dendritic branches by a factor of 11, and the number of glomerular and extraglomerular synapses by factors of 90 and 170, respectively. Each of these parameters increases steadily from birth, in concert with the enlargement of the olfactory mucosa, the overall growth of the brain, and indeed, of the entire animal. We found no evidence of an initial surfeit of processing units, dendritic branches, or synapses. Further elaboration of neural circuitry by each of these measures is also apparent from the time of sexual maturity until the animals reach their full adult size at about 10-12 weeks of age. The developmental strategy in this part of the mouse brain evidently involves prolonged construction that persists until the growth of the body is complete. This ongoing elaboration of neural circuitry in the postnatal mammalian brain may be relevant to understanding a number of unexplained developmental phenomena, including critical periods, the ability of the juvenile brain to recover from injuries that would cause severe and permanent deficits in older animals, and the special ability of the maturing brain to encode large amounts of new information.


The Measurement of Eye Movements in Mild Traumatic Brain Injury: A Structured Review of an Emerging Area.
Mild traumatic brain injury (mTBI), or concussion, occurs following a direct or indirect force to the head that causes a change in brain function. Many neurological signs and symptoms of mTBI can be subtle and transient, and some can persist beyond the usual recovery timeframe, such as balance, cognitive or sensory disturbance that may pre-dispose to further injury in the future. There is currently no accepted definition or diagnostic criteria for mTBI and therefore no single assessment has been developed or accepted as being able to identify those with an mTBI. Eye-movement assessment may be useful, as specific eye-movements and their metrics can be attributed to specific brain regions or functions, and eye-movement involves a multitude of brain regions. Recently, research has focused on quantitative eye-movement assessments using eye-tracking technology for diagnosis and monitoring symptoms of an mTBI. However, the approaches taken to objectively measure eye-movements varies with respect to instrumentation, protocols and recognition of factors that may influence results, such as cognitive function or basic visual function. This review aimed to examine previous work that has measured eye-movements within those with mTBI to inform the development of robust or standardized testing protocols. Medline/PubMed, CINAHL, PsychInfo and Scopus databases were searched. Twenty-two articles met inclusion/exclusion criteria and were reviewed, which examined saccades, smooth pursuits, fixations and nystagmus in mTBI compared to controls. Current methodologies for data collection, analysis and interpretation from eye-tracking technology in individuals following an mTBI are discussed. In brief, a wide range of eye-movement instruments and outcome measures were reported, but validity and reliability of devices and metrics were insufficiently reported across studies. Interpretation of outcomes was complicated by poor study reporting of demographics, mTBI-related features (e.g., time since injury), and few studies considered the influence that cognitive or visual functions may have on eye-movements. The reviewed evidence suggests that eye-movements are impaired in mTBI, but future research is required to accurately and robustly establish findings. Standardization and reporting of eye-movement instruments, data collection procedures, processing algorithms and analysis methods are required. Recommendations also include comprehensive reporting of demographics, mTBI-related features, and confounding variables.


Synthesis and Characterization of Inhalable Flavonoid Nanoparticle for Lung Cancer Cell Targeting.
Current cancer treatments are not adequate to cure cancer disease, as most chemotherapeutic drugs do not differentiate between cancerous and non-cancerous cells; which lead to systemic toxicity and adverse effects. We have developed a promising approach to deliver a potential anti-cancer compound (curcumin) for lung cancer treatment through pulmonary delivery. Three different sizes of curcumin micellar nanoparticles (Cur-NPs) were fabricated and their cytotoxicity effects (proliferation, apoptosis, cell cycle progression) were evaluated against non-small-cell lung cancer, human lung carcinoma (A549) and human lung adenocarcinoma (Calu-3). The in vitro cytotoxicity assay showed that Cur-NPs were more effective to kill lung cancer cells compared to DMSO-solubilised raw curcumin. The potency of the anti-cancer killing activities was size-dependent. Both raw curcumin and Cur-NPs were not toxic to healthy lung cells (BEAS-2B). Smaller Cur-NPs accumulated within nucleus, membrane and cytoplasm. Cur-NPs also induced apoptosis and caused G2/M arrest in both A549 and Calu-3 cell lines. Compared to raw curcumin, Cur-NPs were more effective in suppressing the expression of the inflammatory marker, Interleukin-8 (IL8). The aerosol performance of Cur-NPs was characterized using the next generation impactor (NGI). All Cur-NPs showed promising aerosolization property with mass median aerodynamic diameter (MMAD) and geometric standard deviation (GSD) ranging between 4.8-5.2 and 2.0-2.1, respectively. This study suggests that inhaled curcumin nanoparticles could potentially be used for lung cancer treatment with minimal side effects.


Does prenatal exposure to vitamin D-fortified margarine and milk alter birth weight? A societal experiment.
The present study examined whether exposure to vitamin D from fortified margarine and milk during prenatal life influenced mean birth weight and the risk of high or low birth weight. The study was based on the Danish vitamin D fortification programme, which was a societal intervention with mandatory fortification of margarine during 1961-1985 and voluntary fortification of low-fat milk between 1972 and 1976. The influence of prenatal vitamin D exposure on birth weight was investigated among 51 883 Danish children, by comparing birth weight among individuals born during 2 years before or after the initiation and termination of vitamin D fortification programmes. In total, four sets of analyses were performed. Information on birth weight was available in the Copenhagen School Health Record Register for all school children in Copenhagen. The mean birth weight was lower among the exposed than non-exposed children during all study periods (milk initiation - 20·3 (95 % CI - 39·2, - 1·4) g; milk termination - 25·9 (95 % CI - 46·0, - 5·7) g; margarine termination - 45·7 (95 % CI - 66·6, - 24·8) g), except during the period around the initiation of margarine fortification, where exposed children were heavier than non-exposed children (margarine initiation 27·4 (95 % CI 10·8, 44·0) g). No differences in the odds of high (>4000 g) or low ( < 2500 g) birth weight were observed between the children exposed and non-exposed to vitamin D fortification prenatally. Prenatal exposure to vitamin D from fortified margarine and milk altered birth weight, but the effect was small and inconsistent, reaching the conclusion that vitamin D fortification seems to be clinically irrelevant in relation to fetal growth.


Lesion network localization of free will.
Our perception of free will is composed of a desire to act (volition) and a sense of responsibility for our actions (agency). Brain damage can disrupt these processes, but which regions are most important for free will perception remains unclear. Here, we study focal brain lesions that disrupt volition, causing akinetic mutism (n = 28), or disrupt agency, causing alien limb syndrome (n = 50), to better localize these processes in the human brain. Lesion locations causing either syndrome were highly heterogeneous, occurring in a variety of different brain locations. We next used a recently validated technique termed lesion network mapping to determine whether these heterogeneous lesion locations localized to specific brain networks. Lesion locations causing akinetic mutism all fell within one network, defined by connectivity to the anterior cingulate cortex. Lesion locations causing alien limb fell within a separate network, defined by connectivity to the precuneus. Both findings were specific for these syndromes compared with brain lesions causing similar physical impairments but without disordered free will. Finally, our lesion-based localization matched network localization for brain stimulation locations that disrupt free will and neuroimaging abnormalities in patients with psychiatric disorders of free will without overt brain lesions. Collectively, our results demonstrate that lesions in different locations causing disordered volition and agency localize to unique brain networks, lending insight into the neuroanatomical substrate of free will perception.


The effect of normal aging on the coupling of neural activity to the bold hemodynamic response.
The use of functional neuroimaging to test hypotheses regarding age-related changes in the neural substrates of cognitive processes relies on assumptions regarding the coupling of neural activity to neuroimaging signal. Differences in neuroimaging signal response between young and elderly subjects can be mapped directly to differences in neural response only if such coupling does not change with age. Here we examined spatial and temporal characteristics of the BOLD fMRI hemodynamic response in primary sensorimotor cortex in young and elderly subjects during the performance of a simple reaction time task. We found that 75% of elderly subjects (n = 20) exhibited a detectable voxel-wise relationship with the behavioral paradigm in this region as compared to 100% young subjects (n = 32). The median number of suprathreshold voxels in the young subjects was greater than four times that of the elderly subjects. Young subjects had a slightly greater signal:noise per voxel than the elderly subjects that was attributed to a greater level of noise per voxel in the elderly subjects. The evidence did not support the idea that the greater head motion observed in the elderly was the cause of this greater voxel-wise noise. There were no significant differences between groups in either the shape of the hemodynamic response or in its the within-group variability, although the former evidenced a near significant trend. The overall finding that some aspects of the hemodynamic coupling between neural activity and BOLD fMRI signal change with age cautions against simple interpretations of the results of imaging studies that compare young and elderly subjects.


Treatments for Ocular Adnexal Lymphoma: A Report by the American Academy of Ophthalmology.
To review the literature to determine the efficacy of available treatments for ocular adnexal lymphoma (OAL) and to evaluate the outcomes and complications of treatments in patients older than 13 years. A literature search was conducted last in March 2017 in the PubMed and Cochrane Library databases for English-language original research investigations that evaluated treatment outcomes for OAL. The searches identified 307 unique citations, and 27 studies were selected according to the criteria outlined for this assessment. The 27 studies reviewed comprised 2009 patients. Seventy-five percent of the cases reported were extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT). Twenty-five studies reported results using radiotherapy with or without chemotherapy or surgery. The local control rate of MALT lymphomas with treatments involving radiotherapy averaged 95.9%. Distant and local relapses can occur, and in studies reporting only on MALT lymphomas (884 patients), the 5-year and 10-year disease-free survival rates were reported to be 86.4% and 78.7%, respectively. However, overall survival in patients receiving radiotherapy remained very good, with the 5-year and 10-year survival rates reported to be 93.8% and 84.9%, respectively. Studies that included data on multiple histologic subtypes of lymphoma or non-MALT lymphomas (988 patients) reported local control rates to be 93.1%; 5-year and 10-year disease-free survival rates to be 75.7% and 71.0%, respectively; and 5-year and 10-year overall survival rates to be 78.9% and 73.5%, respectively. Studies on the use of doxycycline for MALT lymphomas (137 patients) reported complete responses of between 4.4% and 13%. Complete and partial responses combined were between 26.7% and 65%. Disease-free survival was not reported for these 2 studies, although progression-free survival was reported to be between 55% and 60.9%. The most frequently reported complications of treatment were cataracts (12.1%) and dry eye (8.5%). For MALT lymphomas, local control, disease-free survival, and overall survival are good with radiation treatment. The results of treatment of non-MALT lymphomas using radiotherapy also were good, but they were not as favorable as the treatment results of MALT lymphomas.


Environmental Noise Annoyance and Mental Health in Adults: Findings from the Cross-Sectional German Health Update (GEDA) Study 2012.
The health implications of environmental noise, especially cardiovascular effects, have been studied intensively. Research on associations between noise and mental health, however, has shown contradictory results. The present study examined associations between individual levels of noise annoyance due to noise from various sources in the living environment and mental health of adults in Germany. It evaluated whether these associations persisted after adjusting for potential covariates. Data were obtained from the cross-sectional "German Health Update" study 2012 (GEDA 2012), a national health interview survey among adults in Germany conducted by the Robert Koch Institute (n = 19,294). Noise annoyance questions referred to overall noise and that from road traffic, neighbours, and air traffic. Mental health was measured with the five-item Mental Health Inventory. Bivariate analysis showed associations between high levels of noise annoyance and impaired mental health for all noise sources except air traffic. After adjusting for covariates (sociodemographic factors, chronic disease, and social support), both men and women who reported high overall noise annoyance showed more than doubled odds of impaired mental health compared to those who were not annoyed. The odds of impaired mental health in the highest noise annoyance category from road traffic and neighbours were also significantly increased. These findings indicate that high noise annoyance is associated with impaired mental health and that this association can vary with the source of environmental noise. Further research on covariates of this association is necessary. Particularly, longitudinal data are required to establish the direction of associations and to address questions of causality.


Sexual risk behaviors among youth heads of household in Gikongoro, south province of Rwanda.
As a result of the 1994 genocide and AIDS, Rwanda has a crisis of orphans. In 2005, the Ministry of Local Governance and Social Affairs of Rwanda has reported one million vulnerable children. Many of these are not only orphans but also youth heads of households (YHH). The purpose of this study was to: (a) identify risk behaviors that expose YHH to HIV infection, (b) determine gender-specific high risk profiles and, (c) determine predictors of sexual onset. A household survey was conducted among 692 YHH, aged 12-24, all beneficiaries of a World Vision basic needs program in Gikongoro, Rwanda, from January to March 2004. Participants were interviewed using a structured questionnaire. Data was collected on socio-demographic variables, HIV/AIDS prevention knowledge and sexual risk behaviors. Bivariate analyses of the study variables were performed to examine differences between males and females. A logistic regression analysis was conducted to analyze factors that were independently associated with the debut of having sex. Forty-one percent of respondents reported sexual onset before age 15. Males were more likely to start earlier than females (50.4% versus 26.7%) but females reported more sexual onset with an older partner. Fifty-eight percent of females had their first intercourse with a partner who was four or more years older than themselves. While sexual activity was low (1.75 mean lifetime sexual partner, 0.45 mean sexual partner last twelve months), sexual experience was related to less social connectedness and use of drugs. Having a close friend also appeared to be protective for sexual debut. The analysis also found that although YHH were aware of some prevention measures against HIV/AIDS, there was low (19.8%) knowledge of the "ABC" prevention program promoted by the government. In addition, despite 85% of respondents knowing someone who had died of AIDS, only 31% perceived themselves at risk of HIV infection, and there was very low (13.2%) condom use among the sexually experienced. Results suggest the urgent need of HIV prevention programs tailored to YHH that provide knowledge, enhance negotiations skills, and increase the perception of HIV infection risk among YHH in Rwanda.


Long-term sobriety strategies for men with co-occurring disorders.
Roughly half of people with severe mental disorders also experience a co-occurring substance use disorder, and recovery from both is a critical objective for health care services. While understanding of abstinence initiation has grown, the strategies people with co-occurring disorders use to maintain sobriety are largely unknown. This article reports strategies for relapse prevention as described by men with co-occurring disorders who achieved one or more years of sobriety. We analyzed semi-structured interviews conducted with a sample of 12 men with co-occurring psychosis and substance use disorder who achieved and maintained sobriety for at least one year, supplemented with demographic and diagnostic clinical record data. These men were participating in residential or outpatient treatment at a private, nonprofit integrated treatment clinic. The 12 men were primarily Caucasian (91.7%) and unmarried (100%), and their ages ranged from 23 to 42 years. The two most common psychiatric disorders were schizoaffective disorder (n = 4, 33.3%) and bipolar disorder (n = 4, 33.3%), while the two most commonly misused substances were alcohol and cannabis. Qualitative analyses showed that participants maintained sobriety for at least one year by building a supportive community, engaging in productive activities, and carefully monitoring their own attitudes toward substances, mental health, and responsibility. Alcoholics Anonymous might act as a catalyst for building skills. People with co-occurring disorders who achieve sobriety use a variety of self-management strategies to prevent relapse-seeking support, activities, and a healthy mindset. The findings suggest a relapse prevention model that focuses on social networks, role functioning, and self-monitoring and conceptualizes self-care as critical to extending periods of wellness.


The processing of feature discontinuities for different cue types in primary visual cortex.
This study examines whether neurons in the primary visual cortex (V1) of the cat (also referred to as area 17) are sensitive to boundaries that are delineated by a difference in features other than luminance contrast. Most research on this issue has concentrated on the responses to texture borders (e.g. 'illusory contours') and has found neurons that are sensitive to such borders in V2 and to a lesser extent in V1. Here neurons in cat area 17 (V1) were exposed to borders that were oblique to the orientation preference of the neuron and that were created by differences in phase, orientation or direction of motion of two drifting sinewave gratings. Nearby phase borders evoked increased firing in 15 out of 98 neurons, orientation borders in 18 out of 98, and direction borders in 15 out of 70 neurons recorded in area 17 (V1) of anesthetized cats. The firing rates of these neurons were enhanced when a feature border was presented close to their receptive field, partly independent of the cue involved. Control experiments with a contrast border showed that the enhanced firing was due to a release of suppression rather than facilitation. A conceptual model is presented that can describe the data and uncovers a peculiarity of the phase domain compared to the orientation and direction domain. The model unifies the knowledge gained here about orientation-specific center-surround interactions, contextual effects, and end-stopping. The data and model suggest that these phenomena are part of a single mechanism that enables the brain to detect feature discontinuities across a range of features.


Inhibition of oxidative stress and cytokine activity by curcumin in amelioration of endotoxin-induced experimental hepatoxicity in rodents.
The present study is aimed at investigating the effect of curcumin (CMN) in salvaging endotoxin-induced hepatic dysfunction and oxidative stress in the liver of rodents. Hepatotoxicity was induced by administering lipopolysaccharide (LPS) in a single dose of 1 mg/kg intraperitoneally to the animals, which were being treated with CMN daily for 7 days. Liver enzymes serum alanine aminotransferase (ALT), serum aspartate aminotransferase (AST) and alkaline phosphatase (ALP), total bilirubin and total protein were estimated in serum. Oxidative stress in liver tissue homogenates was estimated by measuring thiobarbituric acid reactive substances (TBARS), glutathione (GSH) content and superoxide dismutase (SOD) activity. Serum and tissue nitrite was estimated using Greiss reagent and served as an indicator of NO production. A separate set of experiments was performed to estimate the effect of CMN on cytokine levels in mouse serum after LPS challenge. LPS induced a marked hepatic dysfunction evident by rise in serum levels of ALT, AST, ALP and total bilirubin (P < 0.05). TBARS levels were significantly increased, whereas GSH and SOD levels decreased in the liver homogenates of LPS-challenged rats. CMN administration attenuated these effects of LPS successfully. Further CMN treatment also regressed various structural changes induced by LPS in the livers of rats and decreased the levels of tumour necrosis factor-alpha and interleukin-6 in mouse plasma. In conclusion, these findings suggest that CMN attenuates LPS-induced hepatotoxicity possibly by preventing cytotoxic effects of NO, oxygen free radicals and cytokines.


A comparison between performance on selected directions of the star excursion balance test and the Y balance test.
The Star Excursion Balance Test (SEBT) is a widely accepted method of assessing dynamic postural stability. The Y Balance Test (YBT) is a commercially available device for measuring balance that uses 3 (anterior, posteromedial, and posterolateral) of the 8 SEBT directions and has been advocated as a method for assessing dynamic balance. To date, no studies have compared reach performance in these tests in a healthy population. To determine whether any differences exist between reach distance performance for the anterior, posteromedial, and posterolateral directions of the SEBT and the YBT. Descriptive laboratory study. University motion analysis laboratory. A total of 20 healthy active male participants (age = 22.50 ± 3.05 years, height = 1.78 ± 0.82 m, weight = 79.48 ± 11.32 kg, body mass index = 24.96 ± 2.56 kg/m²). Participants carried out 3 trials in each reach direction on each leg on the SEBT and the YBT a minimum of 1 week apart. The means of the 3 trials in each direction on each leg on both tests were calculated. Data were collected after 4 practice trials in each direction. Paired t tests and Bland-Altman plots were used to compare reach distances between the SEBT and the YBT. Participants reached farther in the anterior direction on the SEBT than on the YBT. No differences were observed in the posteromedial and posterolateral directions. Differing postural-control strategies may be used to complete these tasks. This finding has implications for the implementation and interpretation of these dynamic balance tests.


Who decides to have sex? Exploring the perceptions of Timorese women and men through a reproductive justice lens.
Despite national policies to support sexual rights, Timorese women are constrained when making sexual and reproductive health decisions. Contextual understanding of sexual decision making is vital for effective engagement by sexual and reproductive health service providers with communities. An intersectional reproductive justice approach broadens the sexual rights lens allowing for an examination of multi-system factors impacting on sexual rights and health. Using the Matrix of Domination as a conceptual framework, we explored Timorese perceptions around decisions to have sex, and examined intersecting systems of oppression impacting on these decisions. Our study adopted a critical medical anthropological approach using ethnographic methods. A decolonising methodology aimed to make Timorese worldviews central to the analysis. Nine focus group discussions with 80 men and 17 individual reproductive history interviews with women were held in 4 of Timor-Leste's 13 municipalities during October 2015. Findings suggest that decisions to have sex are framed in terms of wishes and rights; however, it was the perceived entitlements of men that were prioritised and predominantly men who made these decisions. Violence, coercion and unwanted pregnancies were linked to decisions about sex, and identified as potential consequences for women, impacting on women's health and sexual rights.


Physical model for predicting grinding rates.
When grinding an optical surface, the material removal rate depends linearly upon pressure and velocity and upon a proportionality constant which accounts for differences in material properties. A physical description of the grinding process has been postulated which results in the linear pressure and velocity relationships and which also allows the value of the proportionality constant to be determined from a knowledge of material properties alone. Previous attempts at predicting material removal rely strictly upon experimental data to determine the proportionality constant. The postulated description incorporates the combined effects of many individual abrasive particles to statistically determine the proportionality constant in terms of abrasive size and the breaking stress of the glass. A simple example indicates that the result is consistent with experiment.


The nab-paclitaxel/gemcitabine regimen for patients with refractory advanced pancreatic adenocarcinoma.
The phase III MPACT trial for metastatic pancreatic cancer (PC) showed improved overall survival (OS), progression free survival (PFS) and response rates (RRs) for first-line nab-paclitaxel (Abraxane) and gemcitabine (the AG combination) compared to gemcitabine monotherapy. The safety and efficacy of the AG combination has not been systematically studied as second-line therapy or beyond for metastatic PC. We conducted an IRB-approved retrospective analysis of all patients diagnosed between September 2010 and August 2014 with advanced refractory PC that received combination treatment with AG at our institution. Demographic and survival data were extracted from the registry. Patients received nab-paclitaxel 125 mg/m2 and gemcitabine 1,000 mg/m2 and on days 1, 8 and 15 of a 28-day cycle with subsequent dose modifications based on tolerance. Data on 59 patients was available; the median age was 61; 55% were male; 56% received AG as second line therapy and 44% received it as third-line or beyond. Five (10%) patients had a confirmed partial response (PR), 23 (47%) had stable disease (SD) and 21 (43%) had disease progression as their best response. Among the 31 (52%) patients who received prior gemcitabine, 18 (58%) had clinical benefit; 3 had a PR and 15 had SD. The median OS was 3.9 months and the median progression-free survival was 3 months. Toxicity was similar to what was reported in the MPACT trial. This retrospective analysis suggests that AG is active in PC patients previously treated with either fluoropyrimidine-based therapy or gemcitabine-based therapy with manageable toxicities.


Effect of Ramadan fasting on diabetes mellitus: a population-based study in Qatar.
Over one billion Muslims fast worldwide during the month of Ramadan. Fasting during Ramadan is a radical change in lifestyle for the period of a lunar month, and it might affect the biochemical parameters among diabetic patients. This study aimed to investigate the effect of Ramadan fasting on the blood levels of glucose, glycated hemoglobin (HbA1c), and lipid profile among diabetic patients observing fast during the Ramadan. An observational study recruiting 1301 Muslim diabetic patients above 18 years age was conducted in diabetic outpatient clinic of Hamad General Hospital, Hamad Medical Corporation, and Primary Health Care Center, Qatar, from July 2012 to September 2013. Data on sociodemographic characteristics (age, sex, nationality, marital status, education level, and occupation) and lifestyle habits (smoking and physical activity), blood pressures, and anthropometric measurements were obtained by a face-to-face interview and measurement using a structured questionnaire. Blood samples were collected for testing glucose, glycosylated hemoglobin (HbA1C), lipid profile, urea, and creatinine (by the licensed research assistants). Slightly less than half of the participants were overweight (BMI: 25-29.9). Significantly higher proportion of female participants were obese as compared with male participants (P<0.001). Among both male participants and female participants, the average level of blood glucose, HbA1c, total cholesterol, low-density and high-density lipoprotein cholesterol, triglycerides, albumin, bilirubin, uric acid, and systolic and diastolic blood pressures were significantly lower during the Ramadan as compared with before Ramadan (P<0.001 each). Results revealed that fasting during Ramadan is significantly associated with decrease in blood lipid profile, blood pressures, glucose, and HbA1C level among diabetic patients. Muslim diabetic patients after the consultation of their primary physician can fast during the month of Ramadan and it might be beneficial for their health.


Language and literacy development of deaf and hard-of-hearing children: successes and challenges.
Childhood hearing loss presents challenges to language development, especially spoken language. In this article, we review existing literature on deaf and hard-of-hearing (DHH) children's patterns and trajectories of language as well as development of theory of mind and literacy. Individual trajectories vary significantly, reflecting access to early identification/intervention, advanced technologies (e.g., cochlear implants), and perceptually accessible language models. DHH children develop sign language in a similar manner as hearing children develop spoken language, provided they are in a language-rich environment. This occurs naturally for DHH children of deaf parents, who constitute 5% of the deaf population. For DHH children of hearing parents, sign language development depends on the age that they are exposed to a perceptually accessible 1st language as well as the richness of input. Most DHH children are born to hearing families who have spoken language as a goal, and such development is now feasible for many children. Some DHH children develop spoken language in bilingual (sign-spoken language) contexts. For the majority of DHH children, spoken language development occurs in either auditory-only contexts or with sign supports. Although developmental trajectories of DHH children with hearing parents have improved with early identification and appropriate interventions, the majority of children are still delayed compared with hearing children. These DHH children show particular weaknesses in the development of grammar. Language deficits and differences have cascading effects in language-related areas of development, such as theory of mind and literacy development.


Protocol-based management of older adults with hip fractures in Delhi, India: a feasibility study.
Worldwide hip fractures are projected to increase from 1.7 million in 1990 to 6.3 million in 2050. In India, conservative estimates suggest an annual incidence of 600,000 osteoporotic hip fractures and this is expected to increase significantly due to ageing and increase life expectancy. Protocol-based 'care pathways' for the management of adults, over 60 years of age, with hip fractures in high-income countries has resulted in decreased mortality rates, early hospital discharge, improved quality of life and reduction in healthcare costs. The study objectives are to determine appropriateness, acceptability and feasibility of adopting best-practice guideline or protocol-based care for the management of hip fractures among older adults in India. The study will also identify barriers and facilitators in recruiting patients and retention till the agreed follow-up period. This will be a mixed-methods prospective cohort study. The quantitative data collection will involve recruitment of consecutive patients aged >50 years with an X-ray-confirmed hip fracture admitted in four tertiary care hospitals in Delhi, India, over a 2-month period. The quantitative data will be collected at three points: from patients at admission to hospital, from medical records at discharge and by telephone interviews with patients at 30 days post hip fracture. Qualitative data collection will involve key informant interviews, conducted with clinical leads and focus group discussions, conducted with groups of healthcare providers and patients and/or their carers. COM-B theoretical framework (capability, opportunity, motivation and behaviour) will be used to explore healthcare providers' behaviour in order to facilitate development and implementation of appropriate integrated care pathway for management of older adults with hip fractures in India. The proposed study will identify gaps in best practice in the management of older people with hip fractures in tertiary care hospitals in Delhi and document barriers and facilitators to the implementation of protocol-based care through recording the contextual realities of the health systems and care-seeking behaviours. Insights into these factors will be used to facilitate the development of protocol-based management of older people with hip fractures that is appropriate, context specific and acceptable by stakeholders in a low- and middle-income country setting, such as India.


Home visitors and child health: analysis of selected programs.
The relationships between selected child health outcomes and programmatic interventions using home visitors are analyzed. The following features of seven programs are systematically reported: program characteristics; description of the home visitors; program objectives, sample size, and research design; outcome measures and reported data. A number of issues such as funding and long-term viability, use of professional or paraprofessional visitors, visitor selection and supervision, and evaluation of home visitor programs require clarification and are discussed. It is concluded that home visitor programs can contribute to child health outcomes such as increased birth weight, improved prenatal care, improved maternal-infant interaction, and improved use of community resources. Pediatricians can be supportive of such programs at many levels: becoming aware of the existence and range of services of home visitor programs in their area that serve families with children and referring families to those programs; being available to advise programs that are in the planning stages; providing advocacy at the local, state, and national level for the funding and development of such programs; and taking the initiative to join multidisciplinary efforts to develop new programs.


Responsibility, alcoholism, and liver transplantation.
Many believe that it is morally wrong to give lower priority for a liver transplant to alcoholics with end-stage liver disease than to patients whose disease is not alcohol-related. Presumably, alcoholism is a disease that results from factors beyond one's control and therefore one cannot be causally or morally responsible for alcoholism or the liver failure that results from it. Moreover, giving lower priority to alcoholics unfairly singles them out for the moral vice of heavy drinking. I argue that the etiology of alcoholism may involve enough control for the alcoholic to be responsible for his condition and accordingly have a weaker claim to receive a new liver than someone who acquires the disease through no fault of his own. In addition, I show why it is more plausible to reframe the question of priority in terms of control and responsibility rather than virtue and vice. Given that medical resources like livers are scarce, some people may justifiably be given lower priority than others in receiving these resources.


Germline CDH1 deletions in hereditary diffuse gastric cancer families.
Germline CDH1 point or small frameshift mutations can be identified in 30-50% of hereditary diffuse gastric cancer (HDGC) families. We hypothesized that CDH1 genomic rearrangements would be found in HDGC and identified 160 families with either two gastric cancers in first-degree relatives and with at least one diffuse gastric cancer (DGC) diagnosed before age 50, or three or more DGC in close relatives diagnosed at any age. Sixty-seven carried germline CDH1 point or small frameshift mutations. We screened germline DNA from the 93 mutation negative probands for large genomic rearrangements by Multiplex Ligation-Dependent Probe Amplification. Potential deletions were validated by RT-PCR and breakpoints cloned using a combination of oligo-CGH-arrays and long-range-PCR. In-silico analysis of the CDH1 locus was used to determine a potential mechanism for these rearrangements. Six of 93 (6.5%) previously described mutation negative HDGC probands, from low GC incidence populations (UK and North America), carried genomic deletions (UK and North America). Two families carried an identical deletion spanning 193 593 bp, encompassing the full CDH3 sequence and CDH1 exons 1 and 2. Other deletions affecting exons 1, 2, 15 and/or 16 were identified. The statistically significant over-representation of Alus around breakpoints indicates it as a likely mechanism for these deletions. When all mutations and deletions are considered, the overall frequency of CDH1 alterations in HDGC is approximately 46% (73/160). CDH1 large deletions occur in 4% of HDGC families by mechanisms involving mainly non-allelic homologous recombination in Alu repeat sequences. As the finding of pathogenic CDH1 mutations is useful for management of HDGC families, screening for deletions should be offered to at-risk families.


The landscape context of cereal aphid-parasitoid interactions.
Analyses at multiple spatial scales may show how important ecosystem services such as biological control are determined by processes acting on the landscape scale. We examined cereal aphid-parasitoid interactions in wheat fields in agricultural landscapes differing in structural complexity (32-100% arable land). Complex landscapes were associated with increased aphid mortality resulting from parasitism, but also with higher aphid colonization, thereby counterbalancing possible biological control by parasitoids and lastly resulting in similar aphid densities across landscapes. Thus, undisturbed perennial habitats appeared to enhance both pests and natural enemies. Analyses at multiple spatial scales (landscape sectors of 0.5-6 km diameter) showed that correlations between parasitism and percentage of arable land were significant at scales of 0.5-2 km, whereas aphid densities responded to percentage of arable land at scales of 1-6 km diameter. Hence, the higher trophic level populations appeared to be determined by smaller landscape sectors owing to dispersal limitation, showing the 'functional spatial scale' for species-specific landscape management.


The hepatitis B vaccine protects re-exposed health care workers, but does not provide sterilizing immunity.
Infection with hepatitis B virus (HBV) can be prevented by vaccination with HB surface (HBs) antigen, which induces HBs-specific antibodies and T cells. However, the duration of vaccine-induced protective immunity is poorly defined for health care workers who were vaccinated as adults. We investigated the immune mechanisms (antibody and T-cell responses) of long-term protection by the HBV vaccine in 90 health care workers with or without occupational exposure to HBV, 10-28 years after vaccination. Fifty-nine of 90 health care workers (65%) had levels of antibodies to HBs antigen above the cut-off (>12 mIU/mL) and 30 of 90 (33%) had HBs-specific T cells that produced interferon-gamma. Titers of antibodies to HBs antigen correlated with numbers of HBs-specific interferon-gamma-producing T cells, but not with time after vaccination. Although occupational exposure to HBV after vaccination did not induce antibodies to the HBV core protein (HBcore), the standard biomarker for HBV infection, CD4(+) and CD8(+) T cells against HBcore and polymerase antigens were detected. Similar numbers of HBcore- and polymerase-specific CD4(+) and CD8(+) T cells were detected in health care workers with occupational exposure to HBV and in patients who acquired immunity via HBV infection. Most of the HBcore- and polymerase-specific T cells were CD45RO(+)CCR7(-)CD127(-) effector memory cells in exposed health care workers and in patients with acquired immunity. In contrast, most of the vaccine-induced HBs-specific T cells were CD45RO(-)CCR7(-)CD127(-) terminally differentiated cells. HBs antigen vaccine-induced immunity protects against future infection but does not provide sterilizing immunity, as evidenced by HBcore- and polymerase-specific CD8(+) T cells in vaccinated health care workers with occupational exposure to HBV. The presence of HBcore- and HBV polymerase-specific T-cell responses is a more sensitive indicator of HBV exposure than detection of HBcore-specific antibodies.


Gene expression, synteny, and local similarity in human noncoding mutation rates.
The human genome is organized with regard to many features such as isochores, Giemsa bands, clusters of genes with similar expression patterns, and contiguous regions with shared evolutionary histories (synteny blocks). In addition to these genomic features, it is clear that mutation rates also vary across the human genome. To address how mutation rates and genomic features are related, we analyzed substitution rates at three classes of putatively neutral noncoding sites (nongenic, intronic, and ancestral repeats) in approximately 14 Mb of human-chimpanzee alignments covering human chromosome 7. Patterns of mutation rate variation inferred from substitution rate variation differ among the three site classes. In particular, we find that intronic mutation rates are strongly affected by the breadth of expression of the genes in which they reside, with broadly expressed genes exhibiting low mutation rates, probably as a consequence of the transcription-coupled repair process acting in the germ line. All site classes show significant local similarities in mutation rate at the megabase scale, and regional similarities in nongenic mutation rate covary with blocks of synteny between the human and mouse genomes, indicating that the evolutionary history of a genomic region is an important determinant of mutation rate.