Sanitation of the liberated territories in Croatia after the Storm campaign: the example of Lika-Senj County.
More than 6,000 km2 of the central and southern areas of the Republic of Croatia were liberated by the Storm campaign, August 4 to 7, 1995. On August 8, 1995, the Ministry of Health sent expert teams to the newly liberated areas, to ensure the basic conditions for the displaced persons to return and live there. To describe the public health measures taken we used the examples of Lika-Senj County (3,746 km2 and 71,215 population). By October 1, 1995, 50 bodies, killed in action, had been detected, pathologically and forensically examined, and then buried. Two hundred thirty-three mostly elderly and sick persons were admitted to the reception center in Gospić; 93 of them returned to their homes, 133 were accommodated with their relatives and friends, and 7 were referred for treatment (4 of them died). Two hundred ten cattle carcasses were buried using adequate sanitation measures, whereas 5,575 sheep, 3,138 head of horned cattle, 298 head of hoofed animals, and 300 head of other cattle were caught alive. The procedures of disinfection, disinfestation, and deodorization were performed in 720 apartments, 2 hotels, 3 schools, 1 kindergarten, 1 health care unit, 1 sports hall, and 5 shops. The main water pump was cleared of mines and partially repaired, whereas some 20 water supply facilities were placed under surveillance. Three food depots were found and properly treated. The entire territory was closely observed for possible occurrence of infectious disease.


Clinical manifestations of Rift Valley fever in humans: Systematic review and meta-analysis.
Rift Valley fever (RVF) is an emerging, neglected, mosquito-borne viral zoonosis associated with significant morbidity, mortality and expanding geographical scope. The clinical signs and symptoms in humans are non-specific and case definitions vary. We reviewed and analysed the clinical manifestations of RVF in humans. In this systematic review and meta-analysis we searched on different dates, the Embase (from 1947 to 13th October 2019), Medline (1946 to 14th October 2019), Global Health (1910 to 15th October 2019), and Web of Science (1970 to 15th October 2019) databases. Studies published in English, reporting frequency of symptoms in humans, and laboratory confirmed RVF were included. Animal studies, studies among asymptomatic volunteers, and single case reports for which a proportion could not be estimated, were excluded. Quality assessment was done using a modified Hoy and Brooks et al tool, data was extracted, and pooled frequency estimates calculated using random effects meta-analysis. Of the 3765 articles retrieved, less than 1% (32 articles) were included in the systematic review and meta-analysis. Nine RVF clinical syndromes were reported including the general febrile, renal, gastrointestinal, hepatic, haemorrhagic, visual, neurological, cardio-pulmonary, and obstetric syndromes. The most common clinical manifestations included fever (81%; 95% Confidence Interval (CI) 69-91; [26 studies, 1286 patients]), renal failure (41%; 23-59; [4, 327]), nausea (38%; 12-67; [6, 325]), jaundice (26%; 16-36; [15, 393]), haemorrhagic disease (26%; 17-36; [16, 277]), partial blindness (24%; 7-45; [11, 225]), encephalitis (21%; 11-33; [4, 327]), cough (4%; 0-17; [4, 11]), and miscarriage (54%) respectively. Death occurred in 21% (95% CI 14-29; [16 studies, 328 patients]) of cases, most of whom were hospitalised. This study delineates the complex symptomatology of human RVF disease into syndromes. This approach is likely to improve case definitions and detection rates, impact outbreak control, increase public awareness about RVF, and subsequently inform 'one-health' policies. This study provides a pooled estimate of the proportion of RVF clinical manifestations alongside a narrative description of clinical syndromes. However, most studies reviewed were case series with small sample sizes and enrolled mostly in-patients and out-patients, and captured symptoms either sparsely or using broad category terms.


18F-FDG PET/CT-based early treatment response evaluation of nanoparticle-assisted photothermal cancer therapy.
Within the field of nanoparticle-assisted photothermal cancer therapy, focus has mostly been on developing novel heat-generating nanoparticles with the right optical and dimensional properties. Comparison and evaluation of their performance in tumor-bearing animals are commonly assessed by changes in tumor volume; however, this is usually a late-occurring event. This study implements 2-deoxy-2-[F-18]fluoro-D-glucose positron emission tomography imaging to perform early evaluation of the treatment outcome of photothermal therapy. Silica-gold nanoshells (NS) are administered intravenously to nude mice bearing human neuroendocrine tumor xenografts and the tumors are irradiated by a near-infrared laser. The animals are positron emission tomography scanned with 2-deoxy-2-[F-18]fluoro-D-glucose one day before and one day after treatment. Using this setup, a significant decrease in tumor uptake of 2-deoxy-2-[F-18]fluoro-D-glucose is found already one day after therapy in the group receiving NS and laser treatment compared to control animals. At this time point no change in tumor volume can be detected. Moreover, the change in tumor uptake, is used to stratify the animals into responders and non-responders, where the responding group matched improved survival. Overall, these findings support the use of 2-deoxy-2-[F-18]fluoro-D-glucose positron emission tomography imaging for preclinical and clinical evaluation and optimization of photothermal therapy.


Intratumoral toremifene therapy and tissue distribution in the baboon.
The purpose of the present study was to evaluate the tissue distribution of toremifene (TOR) in baboons following intra-tissue injections and to examine the effectiveness of intratumoral TOR therapy of baboons with various spontaneous neoplasms. Five healthy baboons (Papio sp.) were used to examine the distribution of TOR following intra-tissue injections. Twenty-three different tissue specimens were collected for HPLC analysis. In addition, four baboons with various spontaneous neoplasms (myxoma, squamous cell carcinoma, lymphosarcoma and adenocarcinoma) were treated with intratumoral TOR and their responses were evaluated. Tissue TOR distribution was also examined in these animals. In the tissue distribution study, target tissue/serum TOR concentration ratios ranged from 138 to 8873 and the target tissue/other tissue ratios ranged from 1.2 to 2428. The distribution of TOR was very favorable, with the highest concentrations outside the injection sites noted in adjacent organs. A marked response was observed in the myxoma and partial responses were observed in the other three cases. Drug level analysis data from these four animals revealed tissue concentrations similar to those seen in the TOR tissue distribution study. Intratumoral administration of TOR can achieve effective local tumor and tissue concentrations, while systemic distribution via circulation to other organs is limited.


Syncytiotrophoblast Extracellular Vesicles from Pre-Eclampsia Placentas Differentially Affect Platelet Function.
Pre-eclampsia (PE) complicates around 3% of all pregnancies and is one of the most common causes of maternal mortality worldwide. The pathophysiology of PE remains unclear however its underlying cause originates from the placenta and manifests as raised blood pressure, proteinuria, vascular or systemic inflammation and hypercoagulation in the mother. Women who develop PE are also at significantly higher risk of subsequently developing cardiovascular (CV) disease. In PE, the failing endoplasmic reticulum, oxidative and inflammatory stressed syncytiotrophoblast layer of the placenta sheds increased numbers of syncytiotrophoblast extracellular vesicles (STBEV) into the maternal circulation. Platelet reactivity, size and concentration are also known to be altered in some women who develop PE, although the underlying reasons for this have not been determined. In this study we show that STBEV from disease free placenta isolated ex vivo by dual placental perfusion associate rapidly with platelets. We provide evidence that STBEV isolated from normal placentas cause platelet activation and that this is increased with STBEV from PE pregnancies. Furthermore, treatment of platelets with aspirin, currently prescribed for women at high risk of PE to reduce platelet aggregation, also inhibits STBEV-induced reversible aggregation of washed platelets. Increased platelet reactivity as a result of exposure to PE placenta derived STBEVs correlates with increased thrombotic risk associated with PE. These observations establish a possible direct link between the clotting disturbances of PE and dysfunction of the placenta, as well as the known increased risk of thromboembolism associated with this condition.


Digital inclusion and functional capacity of older adults living in Florianópolis, Santa Catarina, Brazil (EpiFloripa 2009-2010).
To study the relationship between digital inclusion, represented by the exchange of online messages, and functional capacity of older adults living in Florianópolis, Santa Catarina, Brazil. Data were from EpiFloripa Idoso, a cross-sectional household-based survey conducted between 2009 and 2010 with older adults (60+ years). Functional capacity was represented by the inability / difficulty to perform basic / instrumental activities of daily living, and formed the outcome variable called functional dependence. The main independent variable was self-reported ability to send and receive online messages using the computer. Prevalence ratios (PR) and 95% confidence intervals (95%CI) were estimated with multivariable Poisson regression models. The sample consisted of 1,656 older adults between 60 and 102 years old with a mean age of 70.39 years (SD = 7.79). Older adults who could send and receive online messages without difficulty demonstrated a significant lower prevalence of moderate / severe functional dependence (PR = 0.61; 95%CI: 0.40 - 0.94) after adjustment for demographic, socioeconomic, health and behavioral factors. The exchange of online messages has a strong association with functional independence. The causality of this association cannot be determined. Studies support the hypothesis that the exchange of online messages and functional independence have a bidirectional, additive and synergistic association. Longitudinal studies could elucidate the mechanisms involved in this association, in order to support digital inclusion policies for older adults, and to identify the profile of older adults who would benefit the most from digital inclusion.


Immigration Status, Ethnicity, and Long-term Outcomes following Ischemic Stroke.
To assess the association between immigration status and ethnicity and the outcomes of mortality and vascular event recurrence following ischemic stroke in Ontario, Canada. We conducted a retrospective cohort study using linked administrative and clinical registry-based data from 2002 to 2018 and compared hazards of all-cause mortality and vascular event recurrence in immigrants and long-term residents using inverse probability of treatment weighting accounting for age, sex, income and comorbidities. We stratified analyses by age (≤ 75 and > 75 years) and used interaction terms to evaluate if the association between immigration status and outcomes varied with age or ethnicity. We followed 31,918 adult patients, of whom 2740 (8.6%) were immigrants, for a median follow-up of 5 years. Immigrants had a lower mortality than long-term residents (46.1% vs. 64.5%) which was attenuated after adjustment (hazard ratio 0.94; 95% confidence interval 0.88-1.00), but persisted in those aged under 75 years (HR 0.82; 0.74-0.91). Compared to their respective ethnic long-term resident counterparts, the adjusted hazard of death was higher in South Asian immigrants, similar in Chinese immigrants, and lower in other immigrants (P value for interaction = 0.003). The adjusted hazard of vascular event recurrence (HR 1.01; 0.92-1.11) was similar in immigrants and long-term residents, and this observation persisted across all age and ethnic groups. Long-term mortality following ischemic stroke is lower in immigrants and long-term residents, but is similar after adjustment of baseline characteristics, and it is modified by age at the time of stroke and by ethnicity.


Single-prolonged stress induces apoptosis in dorsal raphe nucleus in the rat model of posttraumatic stress disorder.
Post-traumatic stress disorder (PTSD) is an anxiety disorder that develops after exposure to a life-threatening traumatic experience. Meta-analyses of the brainstem showed that midsagittal area of the pons was significantly reduced in patients with PTSD, suggesting a potential apoptosis in dorsal raphe nucleus after single-prolonged stress (SPS). The aim of this study is to investigate whether SPS induces apoptosis in dorsal raphe nucleus in PTSD rats, which may be a possible mechanism of reduced volume of pons and density of gray matter. In this study, rats were randomly divided into 1d, 7d and 14d groups after SPS along with the control group. The apoptosis rate was determined using annexin V-FITC/PI double-labeled flow cytometry (FCM). Levels of Cytochrome c (Cyt-C) was examined by Western blotting. Expression of Cyt-C on mitochondria in the dorsal raphe nucleus neuron was determined by enzymohistochemistry under transmission electron microscopy (TEM). The change of thiamine monophosphatase (TMP) levels was assessed by enzymohistochemistry under light microscope and TEM. Morphological changes of the ultrastructure of the dorsal raphe nucleus neuron were determined by TEM. Apoptotic morphological alterations were observed in dorsal raphe nucleus neuron for all SPS-stimulate groups of rats. The apoptosis rates were significantly increased in dorsal raphe nucleus neuron of SPS rats, along with increased release of cytochrome c from the mitochondria into the cytoplasm, increased expression of Cyt-C and TMP levels in the cytoplasm, which reached to the peak of increase 7 days of SPS. The results indicate that SPS induced Cyt-C released from mitochondria into cytosol and apoptosis in dorsal raphe nucleus neuron of rats. Increased TMP in cytoplasm facilitated the clearance of apoptotic cells. We propose that this presents one of the mechanisms that lead to reduced volume of pons and gray matter associated with PTSD.


High-resolution mapping of the protein interaction network for the human transcription machinery and affinity purification of RNA polymerase II-associated complexes.
Thirty years of research on gene transcription has uncovered a myriad of factors that regulate, directly or indirectly, the activity of RNA polymerase II (RNAPII) during mRNA synthesis. Yet many regulatory factors remain to be discovered. Using protein affinity purification coupled to mass spectrometry (AP-MS), we recently unraveled a high-density interaction network formed by RNAPII and its accessory factors from the soluble fraction of human cell extracts. Validation of the dataset using a machine learning approach trained to minimize the rate of false positives and false negatives yielded a high-confidence dataset and uncovered novel interactors that regulate the RNAPII transcription machinery, including a new protein assembly we named the RNAPII-Associated Protein 3 (RPAP3) complex.


The Immunomodulatory Signature of Extracellular Vesicles From Cardiosphere-Derived Cells: A Proteomic and miRNA Profiling.
Experimental data demonstrated that the regenerative potential and immunomodulatory capacity of cardiosphere-derived cells (CDCs) is mediated by paracrine mechanisms. In this process, extracellular vesicles derived from CDCs (EV-CDCs) are key mediators of their therapeutic effect. Considering the future applicability of these vesicles in human diseases, an accurate preclinical-to-clinical translation is needed, as well as an exhaustive molecular characterization of animal-derived therapeutic products. Based on that, the main goal of this study was to perform a comprehensive characterization of proteins and miRNAs in extracellular vesicles from porcine CDCs as a clinically relevant animal model. The analysis was performed by identification and quantification of proteins and miRNA expression profiles. Our results revealed the presence of clusters of immune-related and cardiac-related molecular biomarkers in EV-CDCs. Additionally, considering that priming stem cells with inflammatory stimuli may increase the therapeutic potential of released vesicles, here we studied the dynamic changes that occur in the extracellular vesicles from IFNγ-primed CDCs. These analyses detected statistically significant changes in several miRNAs and proteins. Notably, the increase in interleukin 6 (IL6) protein, as well as the increase in mir-125b (that targets IL6 receptor) was especially relevant. These results suggest a potential involvement of EV-CDCs in the regulation of the IL6/IL6R axis, with implications in inflammatory-mediated diseases.


[The lifestyle modification coaching program for secondary stroke prevention].
The purpose of this study was to investigate the effects of the lifestyle modification coaching program on self efficacy, lifestyle and physiologic indexes related to the recurrence of stroke in patients with stroke. Sixty-one patients with stroke registered with a stroke center participated in this study. Of the participants, 32 were assigned to the experimental group and 29 to the control group. Self efficacy, alcohol drinking, smoking, physical activity, BMI (body mass index), WHR (waist-hip ratio), blood lipid level, and blood pressure were measured both for the baseline, as well as after intervention. The lifestyle modification coaching program consisted of an 8-week telecoaching session following face-to-face education. The control group received only the face-to-face education. There were significant differences in physical activities, WHR, blood pressure within and between groups after intervention. The lifestyle modification coaching program had significant influences on blood pressure even after gender, age, and physical activity had been adjusted. The results of the study indicate that lifestyle modification coaching program is effective for physical activity, abdominal obesity and blood pressure. Therefore it can be used by nurses in hospitals and communities as one of the secondary stroke prevention programs for patients with stroke.


Yeast Mph1 helicase dissociates Rad51-made D-loops: implications for crossover control in mitotic recombination.
Eukaryotes possess mechanisms to limit crossing over during homologous recombination, thus avoiding possible chromosomal rearrangements. We show here that budding yeast Mph1, an ortholog of human FancM helicase, utilizes its helicase activity to suppress spontaneous unequal sister chromatid exchanges and DNA double-strand break-induced chromosome crossovers. Since the efficiency and kinetics of break repair are unaffected, Mph1 appears to channel repair intermediates into a noncrossover pathway. Importantly, Mph1 works independently of two other helicases-Srs2 and Sgs1-that also attenuate crossing over. By chromatin immunoprecipitation, we find targeting of Mph1 to double-strand breaks in cells. Purified Mph1 binds D-loop structures and is particularly adept at unwinding these structures. Importantly, Mph1, but not a helicase-defective variant, dissociates Rad51-made D-loops. Overall, the results from our analyses suggest a new role of Mph1 in promoting the noncrossover repair of DNA double-strand breaks.


Diosgenin inhibits atherosclerosis via suppressing the MiR-19b-induced downregulation of ATP-binding cassette transporter A1.
Diosgenin (Dgn), a structural analogue of cholesterol, has been reported to have the hypolipidemic and antiatherogenic properties, but the underlying mechanisms are not fully understood. Given the key roles of macrophages in cholesterol metabolism and atherogenesis, it is critical to investigate macrophage cholesterol efflux and development of atherosclerotic lesion after Dgn treatment. This study was designed to evaluate the potential effects of Dgn on macrophage cholesterol metabolism and the development of aortic atherosclerosis, and to explore its underlying mechanisms. Dgn significantly up-regulated the expression of ATP-binding cassette transporter A1 (ABCA1) protein, but didn't affect liver X receptor α levels in foam cells derived from human THP-1 macrophages and mouse peritoneal macrophages (MPMs) as determined by western blotting. The miR-19b levels were markedly down-regulated in Dgn-treated THP-1 macrophages/MPM-derived foam cells. Cholesterol transport assays revealed that treatment with Dgn alone or together with miR-19b inhibitor notably enhanced ABCA1-dependent cholesterol efflux, resulting in the reduced levels of total cholesterol, free cholesterol and cholesterol ester as determined by high-performance liquid chromatography. The fecal 3H-sterol originating from cholesterol-laden MPMs was increased in apolipoprotein E knockout mice treated with Dgn or both Dgn and antagomiR-19b. Treatment with Dgn alone or together with antagomiR-19b elevated plasma high-density lipoprotein levels, but reduced plasma low-density lipoprotein levels. Accordingly, aortic lipid deposition and plaque area were reduced, and collagen content and ABCA1 expression were increased in mice treated with Dgn alone or together with antagomiR-19b. However, miR-19b overexpression abrogated the lipid-lowering and atheroprotective effects induced by Dgn. The present study demonstrates that Dgn enhances ABCA1-dependent cholesterol efflux and inhibits aortic atherosclerosis progression by suppressing macrophage miR-19b expression.


The role of mothers' and fathers' religiosity in African American adolescents' religious beliefs and practices.
To advance understanding of youth religiosity in its sociocultural context, this study examined the associations between parents' and adolescents' religious beliefs and practices and tested the roles of parent and youth gender and youth ethnic identity in these linkages. The sample included 130 two-parent, African American families. Adolescents (49% female) averaged 14.43 years old. Mothers, fathers, and adolescents were interviewed in their homes about their family and personal characteristics, including their religious beliefs. In a series of 7 nightly phone calls, adolescents reported on their daily practices, including time spent in religious practices (e.g., attending services, prayer), and parents reported on their time spent in religious practices with their adolescents. Findings indicated that mothers' beliefs were linked to the beliefs of sons and daughters, but fathers' beliefs were only associated with the beliefs of sons. Mothers' practices were associated with youths' practices, but the link was stronger when mothers' held moderately strong religious beliefs. Fathers' practices were also linked to youth practices, but the association was stronger for daughters than for sons. Findings highlight the understudied role of fathers in African American families, the importance of examining religiosity as a multidimensional construct, and the utility of ethnic homogeneous designs for illuminating the implications of sociocultural factors in the development of African American youth. (PsycINFO Database Record


A consensual validation of schizoid personality in childhood and adult life.
Life histories and interview descriptions of 23 young men diagnosed as offizoid in middle childhood and adult life and of 20 control subjects not given this diagnosis were presented for diagnostic rating to two independent general psychiatrists. Agreement about the presence or absence of schizoid personality was good. It is concluded that, in the case of boys, the syndrome of schizoid personality in childhood (Asperger's syndrome) corresponds to or is subsumed by the more general clinical picture psychiatrists have of schizoid personality in adult life, and that our original use of this diagnostic label for the youngsters we described was not idiosyncratic.


Enablers and barriers in hospital-to-home transitional care for stroke survivors and caregivers: A systematic review.
To synthesise qualitative research evidence on the experience of stroke survivors and informal caregivers in hospital-to-home transitional care. Due to a shortened hospital stay, stroke survivors/caregivers must take over complex care on discharge from hospital to home. Gaps in the literature warrant a meta-synthesis of qualitative studies on perceived enablers and barriers during this crucial period. A systematic review and meta-synthesis. A review was guided by Enhancing Transparency in Reporting the Synthesis of Qualitative Research (ENTREQ) checklist where six databases were searched from April to June 2020 including CINAHL Plus, MEDLINE, PsycINFO, Scopus, Web of Science and ProQuest and ProQuest Dissertations and Theses. There was no date limit to the search. Selected studies were critically appraised. A thematic synthesis approach was applied. The synthesis of 29 studies identified three major findings. First, partnerships with stroke survivors/caregivers empower discharge preparation, foster competence to navigate health and social care systems and activate self-management capabilities. Second, gaps in discharge planning and the lack of timely postdischarge support contribute to unmet care needs for stroke survivors/caregivers and affect their ability to cope with poststroke changes. Third, stroke survivors/caregivers expect integrated transitional care that promotes shared decision-making and enables long-term self-management at home. Hospital-to-home transition is a challenging period in the trajectory of poststroke rehabilitation and recovery. Further research is required to deepen understandings of all stakeholders' views and address unmet needs during transitional care. Protocols and clinical guidelines relating to discharge planning and transitional care need to be reviewed to ensure partnership approach with survivors/caregivers in the design and delivery of individualised transitional care. Stroke nurses are in a unique position to lead timely support for survivors/caregivers and to bridge service gaps in hospital-to-home transitional care.


Identification of social relation within pedestrian dyads.
This study focuses on social pedestrian groups in public spaces and makes an effort to identify the type of social relation between the group members. As a first step for this identification problem, we focus on dyads (i.e. 2 people groups). Moreover, as a mutually exclusive categorization of social relations, we consider the domain-based approach of Bugental, which precisely corresponds to social relations of colleagues, couples, friends and families, and identify each dyad with one of those relations. For this purpose, we use anonymized trajectory data and derive a set of observables thereof, namely, inter-personal distance, group velocity, velocity difference and height difference. Subsequently, we use the probability density functions (pdf) of these observables as a tool to understand the nature of the relation between pedestrians. To that end, we propose different ways of using the pdfs. Namely, we introduce a probabilistic Bayesian approach and contrast it to a functional metric one and evaluate the performance of both methods with appropriate assessment measures. This study stands out as the first attempt to automatically recognize social relation between pedestrian groups. Additionally, in doing that it uses completely anonymous data and proves that social relation is still possible to recognize with a good accuracy without invading privacy. In particular, our findings indicate that significant recognition rates can be attained for certain categories and with certain methods. Specifically, we show that a very good recognition rate is achieved in distinguishing colleagues from leisure-oriented dyads (families, couples and friends), whereas the distinction between the leisure-oriented dyads results to be inherently harder, but still possible at reasonable rates, in particular if families are restricted to parent-child groups. In general, we establish that the Bayesian method outperforms the functional metric one due, probably, to the difficulty of the latter to learn observable pdfs from individual trajectories.


Localization of a poliovirus type 1 neutralization epitope in viral capsid polypeptide VP1.
Poliovirus type 1 cDNA sequences coding for viral capsid polypeptide VP1 were inserted into the beta-lactamase sequence of Escherichia coli plasmid pBR322. Resulting recombinant plasmid pSW119 expressed in Escherichia coli a VP1-beta-lactamase fusion protein that reacted with antibodies raised against poliovirus capsid polypeptide VP1 and with a monoclonal poliovirus type 1 neutralizing antibody, C3. Deletions of various lengths were generated within the VP1 sequence. The hybrid proteins expressed by the deleted plasmids did not react any more with C3 when the region of VP1 amino acids 95-110 (poliovirus nucleotides 2,754-2,806) was deleted. Therefore, the C3 epitope responsible for virus neutralization is most probably located in this region of the capsid polypeptide.


Autophagy in health and disease: focus on the cardiovascular system.
Autophagy is a highly conserved mechanism of lysosome-mediated protein and organelle degradation that plays a crucial role in maintaining cellular homeostasis. In the last few years, specific functions for autophagy have been identified in many tissues and organs. In the cardiovascular system, autophagy appears to be essential to heart and vessel homeostasis and function; however defective or excessive autophagy activity seems to contribute to major cardiovascular disorders including heart failure (HF) or atherosclerosis. Here, we review the current knowledge on the role of cardiovascular autophagy in physiological and pathophysiological conditions.


A draft genome assembly of the solar-powered sea slug Elysia chlorotica.
Elysia chlorotica, a sacoglossan sea slug found off the East Coast of the United States, is well-known for its ability to sequester chloroplasts from its algal prey and survive by photosynthesis for up to 12 months in the absence of food supply. Here we present a draft genome assembly of E. chlorotica that was generated using a hybrid assembly strategy with Illumina short reads and PacBio long reads. The genome assembly comprised 9,989 scaffolds, with a total length of 557 Mb and a scaffold N50 of 442 kb. BUSCO assessment indicated that 93.3% of the expected metazoan genes were completely present in the genome assembly. Annotation of the E. chlorotica genome assembly identified 176 Mb (32.6%) of repetitive sequences and a total of 24,980 protein-coding genes. We anticipate that the annotated draft genome assembly of the E. chlorotica sea slug will promote the investigation of sacoglossan genetics, evolution, and particularly, the genetic signatures accounting for the long-term functioning of algal chloroplasts in an animal.


Evaluating the utility of an accelerometer and urinary hormone analysis as indicators of estrus in a Zoo-housed koala (Phascolarctos cinereus).
Induced ovulators, such as the koala, do not always have overt signs of estrus, which makes pairing these animals for breeding purposes difficult to achieve in a zoo setting. This study examined the possibility of using alternative methods to behavioral sampling and weight fluctuations for monitoring estrus in a female koala of reproductive age. We attempt to gain an improved understanding of koala estrus and enhance our ability to detect it by combining a noninvasive technique for hormone analysis with a newer method for recording activity level. The findings suggest that activity levels were accurately measured using an accelerometer and increased activity was associated with one estrus behavior, bellowing. Weekly weight declines occur with increases in activity registered from the accelerometer. We also validated an assay to detect progesterone and estradiol in female koala urine in an attempt to detect an estrus spike. However, neither urinary concentrations of estradiol nor progesterone was associated with behavioral cues of estrus in our subject. We suggest that increased female activity, bellowing, and weight loss all indicate a pro-estrus state that requires proper male stimulation in order to achieve a full estrus display.


Comparison of novel risk markers for improvement in cardiovascular risk assessment in intermediate-risk individuals.
Risk markers including coronary artery calcium, carotid intima-media thickness, ankle-brachial index, brachial flow-mediated dilation, high-sensitivity C-reactive protein (CRP), and family history of coronary heart disease (CHD) have been reported to improve on the Framingham Risk Score (FRS) for prediction of CHD, but there are no direct comparisons of these markers for risk prediction in a single cohort. We compared improvement in prediction of incident CHD/cardiovascular disease (CVD) of these 6 risk markers within intermediate-risk participants (FRS >5%-<20%) in the Multi-Ethnic Study of Atherosclerosis (MESA). Of 6814 MESA participants from 6 US field centers, 1330 were intermediate risk, without diabetes mellitus, and had complete data on all 6 markers. Recruitment spanned July 2000 to September 2002, with follow-up through May 2011. Probability-weighted Cox proportional hazard models were used to estimate hazard ratios (HRs). Area under the receiver operator characteristic curve (AUC) and net reclassification improvement were used to compare incremental contributions of each marker when added to the FRS, plus race/ethnicity. Incident CHD defined as myocardial infarction, angina followed by revascularization, resuscitated cardiac arrest, or CHD death. Incident CVD additionally included stroke or CVD death. After 7.6-year median follow-up (IQR, 7.3-7.8), 94 CHD and 123 CVD events occurred. Coronary artery calcium, ankle-brachial index, high-sensitivity CRP, and family history were independently associated with incident CHD in multivariable analyses (HR, 2.60 [95% CI, 1.94-3.50]; HR, 0.79 [95% CI, 0.66-0.95]; HR, 1.28 [95% CI, 1.00-1.64]; and HR, 2.18 [95% CI, 1.38-3.42], respectively). Carotid intima-media thickness and brachial flow-mediated dilation were not associated with incident CHD in multivariable analyses (HR, 1.17 [95% CI, 0.95-1.45] and HR, 0.95 [95% CI, 0.78-1.14]). Although addition of the markers individually to the FRS plus race/ethnicity improved AUC, coronary artery calcium afforded the highest increment (0.623 vs 0.784), while brachial flow-mediated dilation had the least (0.623 vs 0.639). For incident CHD, the net reclassification improvement with coronary artery calcium was 0.659, brachial flow-mediated dilation was 0.024, ankle-brachial index was 0.036, carotid intima-media thickness was 0.102, family history was 0.160 and high-sensitivity CRP was 0.079. Similar results were obtained for incident CVD. Coronary artery calcium, ankle-brachial index, high-sensitivity CRP, and family history were independent predictors of incident CHD/CVD in intermediate-risk individuals. Coronary artery calcium provided superior discrimination and risk reclassification compared with other risk markers.


Designing new biocompatible glass-forming Ti75-x Zr10 Nbx Si15 (x = 0, 15) alloys: corrosion, passivity, and apatite formation.
Glass-forming Ti-based alloys are considered as potential new materials for implant applications. Ti75 Zr10 Si15 and Ti60 Zr10 Nb15 Si15 alloys (free of cytotoxic elements) can be produced as melt-spun ribbons with glassy matrix and embedded single β-type nanocrystals. The corrosion and passivation behavior of these alloys in their homogenized melt-spun states have been investigated in Ringer solution at 37°C in comparison to their cast multiphase crystalline counterparts and to cp-Ti and β-type Ti-40Nb. All tested materials showed very low corrosion rates as expressed in corrosion current densities icorr < 50 nA/cm(2). Electrochemical and surface analytical studies revealed a high stability of the new alloys passive states in a wide potential range. This corresponds to low passive current densities ipass = 2 ± 1 µA/cm(2) based on the growth of oxide films with thickness d <10 nm. A homogeneous constituent distribution in the melt-spun alloys is beneficial for stable surface passivity. The addition of Nb does not only improve the glass-forming ability and the mechanical properties but also supports a high pitting resistance even at extreme anodic polarization up to 4V versus SCE were oxide thickness values of d ∼35 nm are reached. With regard to the corrosion properties, the Nb-containing nearly single-phase glassy alloy can compete with the β-type Ti-40Nb alloy. SBF tests confirmed the ability for formation of hydroxyapatite on the melt-spun alloy surfaces. All these properties recommend the new glass-forming alloys for application as wear- and corrosion-resistant coating materials for implants.


Nursing Leadership Implications for Clinical Placements in Corrections.
Clinical placements in correctional settings offer nursing students unique opportunities for learning mental health and community health concepts, including social justice, restorative justice and the impact of poverty and marginalization on health and life choices. Although there is some evidence to suggest that a small number of nursing programs use clinical placements in correctional settings, relatively little scholarly literature addresses nursing education in such settings, or the implications for nursing leadership when students do have an opportunity to learn in correctional settings. In this paper, we examine the literature that is available on this topic and present the findings of a secondary analysis of interviews with undergraduate nursing students at our nursing program in relation to their clinical placements in correctional settings. Drawing on the students' perspectives, we have found that these placements, in particular, have fostered learning about caring for marginalized populations; themes of hope and restorative justice featured prominently in their descriptions of their learning. Students also emphasized that they learned a great deal about the expanded role of nurses and about caring for marginalized populations. With strong administrative and faculty support, these settings offer students exposure to expert registered nurse mentors who work with clients in an expanded role to facilitate their achievement and stabilization of a broad range of health challenges. They are also role models for students, by showing students that nurses can be agents of hope when working with a diverse client population and their families. We offer recommendations on how to maximize student learning in correctional settings, including a reflection on how to support students' integration of their learning experiences in their nursing practice, with the long-term view that these transformative student experiences have the potential to shape our future nurse leaders.


Designing for action: adapting and implementing a community-based newborn care package to affect national change in Uganda.
There is a lack of literature on how to adapt new evidence-based interventions for maternal and newborn care into local health systems and policy for rapid scale-up, particularly for community-based interventions in low-income settings. The Uganda Newborn Study (UNEST) was a cluster randomised control trial to test a community-based care package which was rapidly taken up at national level. Understanding this process may help inform other studies looking to design and evaluate with scale-up in mind. This study aimed to describe the process of using evidence to design a community-based maternal and newborn care package in rural eastern Uganda, and to determine the dissemination and advocacy approaches used to facilitate rapid policy change and national uptake. We reviewed UNEST project literature including meeting reports and minutes, supervision reports, and annual and midterm reports. National stakeholders, project and district staff were interviewed regarding their role in the study and perceptions of what contributed to uptake of the package under evaluation. Data related to UNEST formative research, study design, implementation and policy influence were extracted and analysed. An advisory committee of key players in development of maternal and newborn policies and programmes in Uganda was constituted from many agencies and disciplines. Baseline qualitative and quantitative data collection was done at district, community and facility level to examine applicability of aspects of a proposed newborn care package to the local setting. Data were summarised and presented to stakeholders to adapt the intervention that was ultimately tested. Quarterly monitoring of key activities and events around the interventions were used to further inform implementation. The UNEST training package, home visit schedule and behaviour change counselling materials were incorporated into the national Village Health Team and Integrated Community Case Management packages while the study was ongoing. Designing interventions for national scale-up requires strategies and planning from the outset. Use of evidence alongside engagement of key stakeholders and targeted advocacy about the burden and potential solutions is important when adapting interventions to local health systems and communities. This approach has the potential to rapidly translate research into policy, but care must be taken not to exceed available evidence while seizing the policy opportunity.


Ensemble modeling of very small ZnO nanoparticles.
The detailed structural characterization of nanoparticles is a very important issue since it enables a precise understanding of their electronic, optical and magnetic properties. Here we introduce a new method for modeling the structure of very small particles by means of powder X-ray diffraction. Using thioglycerol-capped ZnO nanoparticles with a diameter of less than 3 nm as an example we demonstrate that our ensemble modeling method is superior to standard XRD methods like, e.g., Rietveld refinement. Besides fundamental properties (size, anisotropic shape and atomic structure) more sophisticated properties like imperfections in the lattice, a size distribution as well as strain and relaxation effects in the particles and-in particular-at their surface (surface relaxation effects) can be obtained. Ensemble properties, i.e., distributions of the particle size and other properties, can also be investigated which makes this method superior to imaging techniques like (high resolution) transmission electron microscopy or atomic force microscopy, in particular for very small nanoparticles. For the particles under study an excellent agreement of calculated and experimental X-ray diffraction patterns could be obtained with an ensemble of anisotropic polyhedral particles of three dominant sizes, wurtzite structure and a significant relaxation of Zn atoms close to the surface.


Motor planning poststroke: impairment in vector-coded reach plans.
Healthy individuals appear to use both vector-coded reach plans that encode movements in terms of their desired direction and extent, and target-coded reach plans that encode the desired endpoint position of the effector. We examined whether these vector and target reach-planning codes are differentially affected after stroke. Participants with stroke and healthy controls made blocks of reaches that were grouped by target location (providing target-specific practice) and by movement vector (providing vector-specific practice). Reach accuracy was impaired in the more affected arm after stroke, but not distinguishable for target- versus vector-grouped reaches. Reach velocity and acceleration were not only impaired in both the less and more affected arms poststroke, but also not distinguishable for target- versus vector-grouped reaches. As previously reported in controls, target-grouped reaches yielded isotropic (circular) error distributions and vector-grouped reaches yielded error distributions elongated in the direction of the reach. In stroke, the pattern of variability was similar. However, the more affected arm showed less elongated error ellipses for vector-grouped reaches compared to the less affected arm, particularly in individuals with right-hemispheric stroke. The results suggest greater impairment to the vector-coded movement-planning system after stroke, and have implications for the development of personalized approaches to poststroke rehabilitation: Motor learning may be enhanced by practice that uses the preserved code or, conversely, by retraining the more impaired code to restore function.


Do medical procedures in the arm increase the risk of lymphoedema after axillary surgery? A review.
Lymphoedema of the arm is a potentially serious consequence of any axillary procedure performed during the management of breast cancer. In an attempt to reduce its incidence and severity, patients are instructed to avoid venepunctures and blood pressure measurements on the treated arm. These precautions are not possible in some patients and attempts to adhere to them can cause discomfort, anxiety and stress for both patients and their health-care workers. The strength with which these recommendations are made is in contrast to the level of evidence underpinning them. This paper reviews this evidence regarding the safety, or lack thereof, of blood pressure monitoring and intravenous puncture in women who have had axillary surgery. With this evidence generally being anecdotal in nature, there appears to be no rigorous evidence-based support for the risk-reduction behaviours of avoiding blood pressure monitoring and venepuncture in the affected arm in the prevention of lymphoedema after axillary procedure. A clinical trial was proposed to investigate whether such avoidance measures were valuable, but failed during its inception. There remains a need for research from prospective trials on this controversial topic to determine the most appropriate patient recommendations that should be provided after axillary procedure regarding the risks for development of lymphoedema.


Chlorhexidine mouthrinse as an adjunctive treatment for gingival health.
Dental plaque associated gingivitis is a reversible inflammatory condition caused by accumulation and persistence of microbial biofilms (dental plaque) on the teeth. It is characterised by redness and swelling of the gingivae (gums) and a tendency for the gingivae to bleed easily. In susceptible individuals, gingivitis may lead to periodontitis and loss of the soft tissue and bony support for the tooth. It is thought that chlorhexidine mouthrinse may reduce the build-up of plaque thereby reducing gingivitis. To assess the effectiveness of chlorhexidine mouthrinse used as an adjunct to mechanical oral hygiene procedures for the control of gingivitis and plaque compared to mechanical oral hygiene procedures alone or mechanical oral hygiene procedures plus placebo/control mouthrinse. Mechanical oral hygiene procedures were toothbrushing with/without the use of dental floss or interdental cleaning aids and could include professional tooth cleaning/periodontal treatment.To determine whether the effect of chlorhexidine mouthrinse is influenced by chlorhexidine concentration, or frequency of rinsing (once/day versus twice/day).To report and describe any adverse effects associated with chlorhexidine mouthrinse use from included trials. Cochrane Oral Health's Information Specialist searched the following databases: Cochrane Oral Health's Trials Register (to 28 September 2016); the Cochrane Central Register of Controlled Trials (CENTRAL; 2016, Issue 8) in the Cochrane Library (searched 28 September 2016); MEDLINE Ovid (1946 to 28 September 2016); Embase Ovid (1980 to 28 September 2016); and CINAHL EBSCO (Cumulative Index to Nursing and Allied Health Literature; 1937 to 28 September 2016). We searched ClinicalTrials.gov and the World Health Organization International Clinical Trials Registry Platform for ongoing trials. No restrictions were placed on the language or date of publication when searching the electronic databases. We included randomised controlled trials assessing the effects of chlorhexidine mouthrinse used as an adjunct to mechanical oral hygiene procedures for at least 4 weeks on gingivitis in children and adults. Mechanical oral hygiene procedures were toothbrushing with/without use of dental floss or interdental cleaning aids and could include professional tooth cleaning/periodontal treatment. We included trials where participants had gingivitis or periodontitis, where participants were healthy and where some or all participants had medical conditions or special care needs. Two review authors independently screened the search results extracted data and assessed the risk of bias of the included studies. We attempted to contact study authors for missing data or clarification where feasible. For continuous outcomes, we used means and standard deviations to obtain the mean difference (MD) and 95% confidence interval (CI). We combined MDs where studies used the same scale and standardised mean differences (SMDs) where studies used different scales. For dichotomous outcomes, we reported risk ratios (RR) and 95% CIs. Due to anticipated heterogeneity we used random-effects models for all meta-analyses. We included 51 studies that analysed a total of 5345 participants. One study was assessed as being at unclear risk of bias, with the remaining 50 being at high risk of bias, however, this did not affect the quality assessments for gingivitis and plaque as we believe that further research is very unlikely to change our confidence in the estimate of effect. Gingivitis After 4 to 6 weeks of use, chlorhexidine mouthrinse reduced gingivitis (Gingival Index (GI) 0 to 3 scale) by 0.21 (95% CI 0.11 to 0.31) compared to placebo, control or no mouthrinse (10 trials, 805 participants with mild gingival inflammation (mean score 1 on the GI scale) analysed, high-quality evidence). A similar effect size was found for reducing gingivitis at 6 months. There were insufficient data to determine the reduction in gingivitis associated with chlorhexidine mouthrinse use in individuals with mean GI scores of 1.1 to 3 (moderate or severe levels of gingival inflammation). Plaque Plaque was measured by different indices and the SMD at 4 to 6 weeks was 1.45 (95% CI 1.00 to 1.90) standard deviations lower in the chlorhexidine group (12 trials, 950 participants analysed, high-quality evidence), indicating a large reduction in plaque. A similar large reduction was found for chlorhexidine mouthrinse use at 6 months. Extrinsic tooth staining There was a large increase in extrinsic tooth staining in participants using chlorhexidine mouthrinse at 4 to 6 weeks. The SMD was 1.07 (95% CI 0.80 to 1.34) standard deviations higher (eight trials, 415 participants analysed, moderate-quality evidence) in the chlorhexidine mouthrinse group. There was also a large increase in extrinsic tooth staining in participants using chlorhexidine mouthrinse at 7 to 12 weeks and 6 months. Calculus Results for the effect of chlorhexidine mouthrinse on calculus formation were inconclusive. Effect of concentration and frequency of rinsing There were insufficient data to determine whether there was a difference in effect for either chlorhexidine concentration or frequency of rinsing. Other adverse effects The adverse effects most commonly reported in the included studies were taste disturbance/alteration (reported in 11 studies), effects on the oral mucosa including soreness, irritation, mild desquamation and mucosal ulceration/erosions (reported in 13 studies) and a general burning sensation or a burning tongue or both (reported in nine studies). There is high-quality evidence from studies that reported the Löe and Silness Gingival Index of a reduction in gingivitis in individuals with mild gingival inflammation on average (mean score of 1 on the 0 to 3 GI scale) that was not considered to be clinically relevant. There is high-quality evidence of a large reduction in dental plaque with chlorhexidine mouthrinse used as an adjunct to mechanical oral hygiene procedures for 4 to 6 weeks and 6 months. There is no evidence that one concentration of chlorhexidine rinse is more effective than another. There is insufficient evidence to determine the reduction in gingivitis associated with chlorhexidine mouthrinse use in individuals with mean GI scores of 1.1 to 3 indicating moderate or severe levels of gingival inflammation. Rinsing with chlorhexidine mouthrinse for 4 weeks or longer causes extrinsic tooth staining. In addition, other adverse effects such as calculus build up, transient taste disturbance and effects on the oral mucosa were reported in the included studies.


Planning C2 pedicle screw placement with multiplanar reformatted cervical spine computed tomography.
Careful preoperative planning with thin-slice computed tomography (CT) scan is useful for hardware placement at C2. Prior studies have shown considerable variability in the proportion of C2 vertebrae considered safe for pedicle screw placement, depending on the imaging technique used. Our work sought to more carefully define that proportion using a refined imaging technique on a large number of submillimeter CT scans. We reviewed 150 submillimeter cervical spine studies randomly selected from CT scans performed at a Level 1 trauma center. OsiriX™ image analysis software was used to propagate a 5-mm cylinder through the plane of the pedicle on paracoronal reformatted CT scans. Hounsfield unit attenuation was used to determine whether the cylinder violated the pedicle. Binomial data were generated to determine the proportion of pedicles that would allow safe screw placement. We analyzed 300 pedicles in 150 patients. Using a standard C2 pedicle starting point, 32% of pedicles were breached by the 5-mm diameter cylinder. When screw trajectory was adjusted by moving the cylinder to fit the pedicle isthmus, establishing an optimized starting point, only 14% of pedicles were breached. Average pedicle length was 27.3 mm for screws that would have crossed the isthmus versus 13.2 mm for screws that would have stopped short due to potential breach. Findings of the current work suggest that preoperative imaging analysis or navigation can be useful adjuncts when anatomical variants are present.


Confirmation of Sarcocystis jamaicensis Sarcocysts in IFN-γ Gene Knockout Mice Orally Inoculated with Sporocysts from a Red-Tailed Hawk ( Buteo jamaicensis).
Here, we report confirmation of sarcocysts of Sarcocystis jamaicensis in an experimental intermediate host, IFN-γ gene knockout (KO) mice orally inoculated sporocysts from its natural definitive host, a red-tailed hawk ( Buteo jamaicensis) (RTH). A RTH submitted to the Carolina Raptor Center, Huntersville, North Carolina, was euthanized because it could not be rehabilitated and released. Fully sporulated sporocysts from intestinal scrapings of the RTH were orally fed to 2 laboratory-reared outbred Swiss Webster mice (SW; Mus musculus) and to 2 KO mice. The sporocysts were infective for KO mice but not to SW mice. Both SW mice remained asymptomatic, and neither schizonts nor sarcocysts were found in their tissues when euthanized on day 54 post-inoculation (PI). The KO mice developed neurological signs and were necropsied 38-54 days PI. Schizonts/merozoites were found in both KO mice euthanized and they were confined to the brain. The predominant lesion was meningoencephalitis. Microscopic sarcocysts were found in muscles of both KO mice. When viewed with light microscopy, the sarcocyst wall appeared thin (<1 μm thick) and smooth. Ultrastructural details of sarcocysts are described.


Impact of hepatitis C virus infection on the course and outcome of patients with acute alcoholic hepatitis.
Limited information is available on the impact of hepatitis C virus (HCV) infection on the clinical course and outcome of acute alcoholic hepatitis (AH), a condition with a significant mortality. We designed this retrospective study to assess effect of HCV on the outcome of patients with AH. Medical charts of patients with a discharge diagnosis of AH (defined using rigorous clinical criteria) were reviewed. Patients were stratified based on the presence or absence of concomitant HCV infection. The disease severity was estimated at admission and at day 7 using model for end-stage disease and discriminant function index scores. Patient survival at 6 months was confirmed with the county death registry. A total of 76 (29 HCV positive) AH cases were analyzed. At admission, disease severity was similar in both groups with severe disease in 53% (49% of AH alone and 59% of AH+HCV; P = 0.18). Although severity scores at day 7 were not available for all patients, disease severity tended to be worse for patients with AH+HCV. Kaplan-Meier survival curves showed a poor survival for AH+HCV compared with AH alone (69 vs. 91%; log-rank P = 0.015). Although patients with AH+HCV were treated less often compared with AH alone (27 vs. 54%; P = 0.05), HCV emerged as an independent risk factor for a poor outcome at 6 months (Cox proportional hazard ratio 8.45; P = 0.01) after controlling for patient demographics, disease severity at admission, and treatment. HCV may be a risk factor for patients with AH with a worse outcome at 6 months. If our findings are confirmed in larger databases, prospective studies are needed to examine mechanisms for this effect of HCV on the outcome of AH.


Real-time dynamic SERS detection of galectin using glycan-decorated gold nanoparticles.
We present the application of surface-enhanced Raman scattering (SERS) spectroscopy for the fast, sensitive and highly specific detection of the galectin-9 (Gal-9) protein in binding buffer (mimicking natural conditions). The method involves the use of specifically designed nanotags comprising glycan-decorated gold nanoparticles encoded with 4-mercaptobenzoic acid. At fast time scales Gal-9 can be detected down to a concentration of 1.2 nM by monitoring the SERS signal of the reporter, driven by aggregation of the functionalized Au NPs tags, induced by Gal-9 recognition. We additionally demonstrate that the sensitivity and concentration working range of the sensor can be tuned via control of aggregation dynamics and cluster size distribution.


An Analysis of Burnout among Military General Surgery Residents.
Resident burnout is an increasing issue in graduate medical education programs. Military graduate medical education is unique in numerous ways and may have different rates of burnout as well as different causes. This study aims to assess resident burnout rates and contributing factors among military general surgery residents. Using Department of Defense approved software, an anonymous survey was created and distributed to all general surgery residents (n = 180) in 6 US medical centers where there are general surgery residency programs. The survey contained an Abbreviated Maslach Burnout Index questionnaire, multiple choice questions including several military-specific questions, and 2 open ended questions. Rates of burnout and potential risk factors associated with burnout were analyzed. After the collection period, 92 of 180 (51%) residents completed all Abbreviated Maslach Burnout Index questions, demographics, and military specific questions with an opportunity for written comments. Notable demographic findings of the respondents were that 64% were male, 65% were married or engaged, 40% had children, and 69% had no student loan debt. Overall, there was a 66% rate of burnout in any tertile. Variables found to be significant for overall burnout included the likelihood the resident plans to stay beyond their active duty service obligation and the perceived level of autonomy. Of the written responses, the most commonly cited contributing factor was the work burden from nonclinical and/or administrative tasks while the most common protective factor was resident camaraderie. Overall, burnout rates are similar among military general surgery residents compared to published reports of civilians. The close association with resident burnout and anticipation of early withdrawal from military service demonstrates this topic is potentially important to retention of the military medical force. The topics of increased resident autonomy, decreased non-clinical duties, and efforts to increase resident camaraderie should be more closely evaluated.


Essential versus accessory aspects of cell death: recommendations of the NCCD 2015.
Cells exposed to extreme physicochemical or mechanical stimuli die in an uncontrollable manner, as a result of their immediate structural breakdown. Such an unavoidable variant of cellular demise is generally referred to as 'accidental cell death' (ACD). In most settings, however, cell death is initiated by a genetically encoded apparatus, correlating with the fact that its course can be altered by pharmacologic or genetic interventions. 'Regulated cell death' (RCD) can occur as part of physiologic programs or can be activated once adaptive responses to perturbations of the extracellular or intracellular microenvironment fail. The biochemical phenomena that accompany RCD may be harnessed to classify it into a few subtypes, which often (but not always) exhibit stereotyped morphologic features. Nonetheless, efficiently inhibiting the processes that are commonly thought to cause RCD, such as the activation of executioner caspases in the course of apoptosis, does not exert true cytoprotective effects in the mammalian system, but simply alters the kinetics of cellular demise as it shifts its morphologic and biochemical correlates. Conversely, bona fide cytoprotection can be achieved by inhibiting the transduction of lethal signals in the early phases of the process, when adaptive responses are still operational. Thus, the mechanisms that truly execute RCD may be less understood, less inhibitable and perhaps more homogeneous than previously thought. Here, the Nomenclature Committee on Cell Death formulates a set of recommendations to help scientists and researchers to discriminate between essential and accessory aspects of cell death.


Oxidized Docosahexaenoic Acid Species and Lipid Peroxidation Products Increase Amyloidogenic Amyloid Precursor Protein Processing.
One of the main characteristics of Alzheimer's disease (AD) is the β-amyloid peptide (Aβ) generated by β- and γ-secretase processing of the amyloid precursor protein (APP). Previously it has been demonstrated that polyunsaturated fatty acids (PUFAs), especially docosahexaenoic acid (DHA), are associated with a reduced risk of AD caused by decreased Aβ production. However, in epidemiological studies and nutritional approaches, the outcomes of DHA-dependent treatment were partially controversial. PUFAs are very susceptible to reactive oxygen species and lipid peroxidation, which are increased during disease pathology. In line with published results, lipid peroxidation was elevated in human postmortem AD brains; especially 4-hydroxy-nonenal (HNE) was increased. To investigate whether lipid peroxidation is only a consequence or might also influence the processes leading to AD, we analyzed 7 different oxidized lipid species including 5 oxidized DHA derivatives and the lipid peroxidation products of ω-3 and ω-6 PUFAs, HNE and 4-hydroxy-hexenal, in human neuroblastoma cells and mouse mixed cortical neurons. In the presence of oxidized lipids Aβ and soluble β-secreted APP levels were elevated, whereas soluble α-secreted APP was decreased, suggesting a shift from the nonamyloidogenic to the amyloidogenic pathway of APP processing. Furthermore, β- and γ-secretase activity was increased by oxidized lipids via increased gene expression and additionally by a direct effect on β-secretase activity. Importantly, only 1% oxidized DHA was sufficient to revert the protective effect of DHA and to significantly increase Aβ production. Therefore, our results emphasize the need to prevent DHA from oxidation in nutritional approaches and might help explain the divergent results of clinical DHA studies.


Osteopathic postdoctoral training institutions.
The Osteopathic Postdoctoral Training Institution (OPTI) program was established by the American Osteopathic Association (AOA) in 1995 as a support and accreditation structure for osteopathic graduate medical education. Since 1999, when the AOA began to require that all AOA-approved osteopathic internships and residencies take place within this structure, the individual programs have been evolving separately, with little interaction--and with varying degrees of success. The AOA's 2007 Annual OPTI Workshop, scheduled to take place this April, is being organized with the primary goal of improving OPTI collaboration to better define standards for quality and promote program development. In response to common challenges and concerns, workshop participants will also be asked to define and reevaluate internal partnerships and shared goals as components of the larger system.


Evaluation of personal protection methods against phlebotomine sand flies including vectors of leishmaniasis in Panama.
Three personal protection methods were evaluated against phlebotomine sand flies in Panama. Skin applications of five selected repellents including deet (N,N-diethyl-m-toluamide) provided a mean coefficient of protection (CP) of 99.2% against the attack of at least three sand fly species. Deet-treated net jackets also provided good protection, but it was concluded that an additional application of repellent to the unprotected face was necessary for maximum protection. Permethrin-treated clothing did not provide the protection expected. Apparently sand fly behavior and resistance to quick knock-down were responsible for the numbers of bites recorded, and therefore maximum protection from bites would require application of deet or another suitable repellent to the exposed skin when wearing permethrin-treated clothing.


Kinematic analysis of dysphagia: significant parameters of aspiration related to bolus viscosity.
The purpose of this study was to investigate the mechanisms of aspiration with respect to the viscosity of ingested material in patients with dysphagia. Seventy patients with dysphagia underwent videofluoroscopic swallow studies (VFSS) between May 1, 2009 and September 30, 2009. Based on the findings of the VFSS, patients were divided into three groups: a thick-fluid aspiration group, a thin-fluid aspiration group, and a no-aspiration group. Kinematic analyses were performed during thick-fluid swallowing. Among our 70 patients, 23 had thick-fluid aspiration, 20 had thin-fluid aspiration, and 27 had no aspiration. A shortened duration of upper esophageal sphincter (UES) opening, a shorter interval between UES opening and peak pharyngeal constriction, and a diminished extent of laryngeal elevation were all significant risk factors for thick-fluid aspiration. A prolonged latency of the swallowing reflex, pharyngeal transit time, and the interval between bolus arrival at the vallecula and laryngeal elevation were all significant risk factors for thin-fluid aspiration. Our kinematic analysis of dysphagia employing the VFSS indicated that the mechanisms relevant to aspiration differed with respect to food viscosity.


Early nasopharyngeal microbial signature associated with severe influenza in children: a retrospective pilot study.
A few studies have highlighted the importance of the respiratory microbiome in modulating the frequency and outcome of viral respiratory infections. However, there are insufficient data on the use of microbial signatures as prognostic biomarkers to predict respiratory disease outcomes. In this study, we aimed to evaluate whether specific bacterial community compositions in the nasopharynx of children at the time of hospitalization are associated with different influenza clinical outcomes. We utilized retrospective nasopharyngeal (NP) samples (n=36) collected at the time of hospital arrival from children who were infected with influenza virus and had been symptomatic for less than 2 days. Based on their clinical course, children were classified into two groups: patients with mild influenza, and patients with severe respiratory or neurological complications. We implemented custom 16S rRNA gene sequencing, metagenomic sequencing and computational analysis workflows to classify the bacteria present in NP specimens at the species level. We found that increased bacterial diversity in the nasopharynx of children was strongly associated with influenza severity. In addition, patients with severe influenza had decreased relative abundance of Staphylococcus aureus and increased abundance of Prevotella (including P. melaninogenica), Streptobacillus, Porphyromonas, Granulicatella (including G. elegans), Veillonella (including V. dispar), Fusobacterium and Haemophilus in their nasopharynx. This pilot study provides proof-of-concept data for the use of microbial signatures as prognostic biomarkers of influenza outcomes. Further large prospective cohort studies are needed to refine and validate the performance of such microbial signatures in clinical settings.


Human-directed social behaviour in dogs shows significant heritability.
Through domestication and co-evolution with humans, dogs have developed abilities to attract human attention, e.g. in a manner of seeking assistance when faced with a problem solving task. The aims of this study were to investigate within breed variation in human-directed contact seeking in dogs and to estimate its genetic basis. To do this, 498 research beagles, bred and kept under standardized conditions, were tested in an unsolvable problem task. Contact seeking behaviours recorded included both eye contact and physical interactions. Behavioural data was summarized through a principal component analysis, resulting in four components: test interactions, social interactions, eye contact and physical contact. Females scored significantly higher on social interactions and physical contact and age had an effect on eye contact scores. Narrow sense heritabilities (h(2) ) of the two largest components were estimated at 0.32 and 0.23 but were not significant for the last two components. These results show that within the studied dog population, behavioural variation in human-directed social behaviours was sex dependent and that the utilization of eye contact seeking increased with age and experience. Hence, heritability estimates indicate a significant genetic contribution to the variation found in human-directed social interactions, suggesting that social skills in dogs have a genetic basis, but can also be shaped and enhanced through individual experiences. This research gives the opportunity to further investigate the genetics behind dogs' social skills, which could also play a significant part into research on human social disorders such as autism.


Water pipe smoking and its association with cigarette and cannabis use in young adults in Switzerland.
Water pipe is a traditional method of tobacco use, which is epidemically spreading throughout Europe. There are scarce data about the use of water pipe and its relation to other addictive behaviors among young adults in Western countries. It was our aim to identify the sociodemographic characteristics of water pipe users in Switzerland and to describe concurrent cigarette and cannabis smoking habits. Young adults aged 16-30 years were evaluated based on a 16-item standardized questionnaire on tobacco consumption and exhaled carbon monoxide. Current water pipe smoking was defined as water pipe use at least once within the last 4 weeks; regular water pipe smoking was defined as water pipe use at least once a week during the last 52 weeks. Out of 353 volunteers, a total of 204 subjects (mean age 21 ± 3.5 years, 113 males) met the inclusion criteria for the study. A total of 78% (n = 160), 30.0% (n = 55) and 3.9% (n = 8) reported ever, current and regular water pipe smoking, respectively. Males smoked more often than females: 2.8 sessions/year (interquartile range 1.1-8) versus 2 sessions/year (interquartile range 0-4; p = 0.022). The major risk factor for ever smoking water pipe was cigarette smoking (odds ratio 6.22, 95% confidence interval 2.33-16.62), followed by cannabis consumption (odds ratio 1.44, 95% confidence interval 1.29-1.62). Ever water pipe smoking was more common among current cannabis users (100 vs. 0%; p < 0.0001) and related to higher exhaled carbon monoxide values (6.0 ± 9.0 vs. 2.1 ± 4.6 ppm; p < 0.001). Water pipe smoking is common among young adults and strongly associated with cigarette smoking and cannabis consumption.


The physiological variability of channel density in hippocampal CA1 pyramidal cells and interneurons explored using a unified data-driven modeling workflow.
Every neuron is part of a network, exerting its function by transforming multiple spatiotemporal synaptic input patterns into a single spiking output. This function is specified by the particular shape and passive electrical properties of the neuronal membrane, and the composition and spatial distribution of ion channels across its processes. For a variety of physiological or pathological reasons, the intrinsic input/output function may change during a neuron's lifetime. This process results in high variability in the peak specific conductance of ion channels in individual neurons. The mechanisms responsible for this variability are not well understood, although there are clear indications from experiments and modeling that degeneracy and correlation among multiple channels may be involved. Here, we studied this issue in biophysical models of hippocampal CA1 pyramidal neurons and interneurons. Using a unified data-driven simulation workflow and starting from a set of experimental recordings and morphological reconstructions obtained from rats, we built and analyzed several ensembles of morphologically and biophysically accurate single cell models with intrinsic electrophysiological properties consistent with experimental findings. The results suggest that the set of conductances expressed in any given hippocampal neuron may be considered as belonging to two groups: one subset is responsible for the major characteristics of the firing behavior in each population and the other is responsible for a robust degeneracy. Analysis of the model neurons suggests several experimentally testable predictions related to the combination and relative proportion of the different conductances that should be expressed on the membrane of different types of neurons for them to fulfill their role in the hippocampus circuitry.


Th1 cytokines are essential for placental immunity to Listeria monocytogenes.
The fetal allograft poses an immunological challenge: how is it protected while immunity to pathogens, particularly those that replicate in the placenta, is maintained? Several theories have been proposed to explain this fetal protection, including a pregnancy-based bias towards a Th2 rather than Th1 cytokine profile in order to avoid generating cytotoxic T cells that could threaten the fetus. Listeria monocytogenes preferentially replicates in the placenta and systemically requires a Th1 response for sterile eradication. In the placenta, the Th1 cytokines tumor necrosis factor alpha (TNF-alpha) and gamma interferon (IFN-gamma) are also synthesized in response to this pathogen, without fetal loss. Here we show, by using mice homozygous for null mutations in either the cytokine or cytokine receptor genes, a requirement for both TNF-alpha and IFN-gamma signaling for an effective placental immune response to L. monocytogenes. However, T cells were not recruited to the placenta. Genetic studies in which the fetal component of the placenta was genetically different from the mother indicated that both the production of and response to these cytokines were maternal. Despite the requirement for these cytokines, the early recruitment of neutrophils to the placenta was normal. Consequently, the bacterium appeared to be delayed in its colonization of this organ and did not fully gain hold until 72 h postinfection. These data show a requirement for Th1 cytokines during pregnancy for effective immunity and indicate that a bias away from Th1 cytokine synthesis is not a necessary prerequisite of pregnancy.


The crust of the Moon as seen by GRAIL.
High-resolution gravity data obtained from the dual Gravity Recovery and Interior Laboratory (GRAIL) spacecraft show that the bulk density of the Moon's highlands crust is 2550 kilograms per cubic meter, substantially lower than generally assumed. When combined with remote sensing and sample data, this density implies an average crustal porosity of 12% to depths of at least a few kilometers. Lateral variations in crustal porosity correlate with the largest impact basins, whereas lateral variations in crustal density correlate with crustal composition. The low-bulk crustal density allows construction of a global crustal thickness model that satisfies the Apollo seismic constraints, and with an average crustal thickness between 34 and 43 kilometers, the bulk refractory element composition of the Moon is not required to be enriched with respect to that of Earth.


Dietary influences on the hepatic mixed-function oxidase system in the rat after portacaval anastomosis.
Studies were undertaken to determine the influence of diet on the hepatic mixed-function oxidase system in rats after portacaval anastomosis. Male rats fed either a cereal-based or purified diet underwent portacaval anastomosis. Weight gain after surgery was highly dependent on the diet. Because rats fed the cereal-based diet weighed 50% less at 4-5 wk after portacaval shunt surgery than unoperated controls fed the same diet, pair-fed unoperated controls were also studied. Rats fed the purified diet grew normally after shunting and therefore studies with this diet did not require pair-fed controls. Shunted rats fed the cereal-based diet had lower liver weights, hepatic microsomal protein concentrations, ethylmorphine N-demethylase, aniline hydroxylase, and cytochromes P450 and b5 when compared with corresponding values in unoperated controls fed the same diet ad libitum; comparisons with pair-fed controls indicated that decreased intake of the cereal-based diet could account for part but not all of the changes seen after portacaval anastomosis. It was shown in rats fed the purified diet that sham operation had no effect on the mixed-function oxidase system, whereas portacaval anastomosis resulted in reduced liver weight, microsomal protein, ethylmorphine N-demethylase, aniline hydroxylase, and cytochrome P450. In comparing shunted rats fed the two types of diet, in contrast with the effects of these diets in normal rats, these hepatic microsomal mixed-function oxidase system components were generally lower in the shunted rats fed the cereal diet. It is concluded that although decreased mixed-function oxidase activity occurs after portacaval anastomosis in the rat, the diet can have an additional substantial influence. Dietary influences on drug oxidations may be an important consideration in drug therapy after portacaval anastomosis.


Useful Interplay Between Spontaneous ADR Reports and Electronic Healthcare Records in Signal Detection.
Spontaneous reporting systems (SRSs) remain the cornerstone of post-marketing drug safety surveillance despite their well-known limitations. Judicious use of other available data sources is essential to enable better detection, strengthening and validation of signals. In this study, we investigated the potential of electronic healthcare records (EHRs) to be used alongside an SRS as an independent system, with the aim of improving signal detection. A signal detection strategy, focused on a limited set of adverse events deemed important in pharmacovigilance, was performed retrospectively in two data sources-(1) the Exploring and Understanding Adverse Drug Reactions (EU-ADR) database network and (2) the EudraVigilance database-using data between 2000 and 2010. Five events were considered for analysis: (1) acute myocardial infarction (AMI); (2) bullous eruption; (3) hip fracture; (4) acute pancreatitis; and (5) upper gastrointestinal bleeding (UGIB). Potential signals identified in each system were verified using the current published literature. The complementarity of the two systems to detect signals was expressed as the percentage of the unilaterally identified signals out of the total number of confirmed signals. As a proxy for the associated costs, the number of signals that needed to be reviewed to detect one true signal (number needed to detect [NND]) was calculated. The relationship between the background frequency of the events and the capability of each system to detect signals was also investigated. The contribution of each system to signal detection appeared to be correlated with the background incidence of the events, being directly proportional to the incidence in EU-ADR and inversely proportional in EudraVigilance. EudraVigilance was particularly valuable in identifying bullous eruption and acute pancreatitis (71 and 42 % of signals were correctly identified from the total pool of known associations, respectively), while EU-ADR was most useful in identifying hip fractures (60 %). Both systems contributed reasonably well to identification of signals related to UGIB (45 % in EudraVigilance, 40 % in EU-ADR) but only fairly for signals related to AMI (25 % in EU-ADR, 20 % in EudraVigilance). The costs associated with detection of signals were variable across events; however, it was often more costly to detect safety signals in EU-ADR than in EudraVigilance (median NNDs: 7 versus 5). An EHR-based system may have additional value for signal detection, alongside already established systems, especially in the presence of adverse events with a high background incidence. While the SRS appeared to be more cost effective overall, for some events the costs associated with signal detection in the EHR might be justifiable.


Modeling fence location and density at a regional scale for use in wildlife management.
Barbed and woven wire fences, common structures across western North America, act as impediments to wildlife movements. In particular, fencing influences pronghorn (Antilocapra americana) daily and seasonal movements, as well as modifying habitat selection. Because of fencing's impacts to pronghorn and other wildlife, it is a potentially important factor in both wildlife movement and habitat selection models. At this time, no geospatial fencing data is available at regional scales. Consequently, we constructed a regional fence model using a series of land tenure assumptions for the Hi-Line region of northern Montana--an area consisting of 13 counties over 103,400 km(2). Randomized 3.2 km long transects (n = 738) on both paved and unpaved roads were driven to collect information on habitat, fence densities and fence type. Using GIS, we constructed a fence location and a density model incorporating ownership, size, neighboring parcels, township boundaries and roads. Local knowledge of land ownership and land use assisted in improving the final models. We predict there is greater than 263,300 km of fencing in the Hi-Line region, with a maximum density of 6.8 km of fencing per km(2) and mean density of 2.4 km of fencing per km(2). Using field data to assess model accuracy, Cohen's Kappa was measured at 0.40. On-the-ground fence modification or removal could be prioritized by identifying high fence densities in critical wildlife areas such as pronghorn migratory pathways or sage grouse lekking habitat. Such novel fence data can assist wildlife and land managers to assess effects of anthropogenic features to wildlife at various scales; which in turn may help conserve declining grassland species and overall ecological functionality.


Evaluation of Validity Evidence for Personality, Emotional Intelligence, and Situational Judgment Tests to Identify Successful Residents.
The ability to identify candidates who will thrive and successfully complete their residency is especially critical for general surgery programs. To assess the extent to which 3 screening tools used extensively in industrial selection settings-emotional intelligence (EQ), personality profiles, and situational judgment tests (SJTs)-could identify successful surgery residents. In this analysis, personality profiles, EQ assessments, and SJTs were administered from July through August 2015 to 51 postgraduate year 1 through 5 general surgery residents in a large general surgery residency program. Associations between these variables and residency performance were investigated through correlation and hierarchical regression analyses. Completion of EQ, personality profiles, and SJT assessments. Performance in residency as measured by a comprehensive performance metric. A score of zero represented a resident whose performance was consistent with that of their respective cohort's performance; below zero, worse performance; and greater than zero, better performance. Of the 61 eligible residents, 51 (84%) chose to participate and 22 (43%) were women. US Medical Licensing Examination Step 1 (USMLE1), but not USMLE2, emerged as a significant factor (t2,49 = 1.98; β = 0.30; P = .03) associated with overall performance. Neither EQ facets nor overall EQ offered significant incremental validity over USMLE1 scores. Inclusion of the personality factors did not significantly alter the test statistic and did not explain any additional portion of the variance. By contrast, inclusion of SJT scores accounted for 15% more of the variance than USMLE1 scores alone, resulting in a total of 25% of the variance explained by both USMLE1 and SJT scores (F2,57 = 7.47; P = .002). Both USMLE1 (t = 2.21; P = .03) and SJT scores (t = 2.97; P = .005) were significantly associated with overall resident performance. This study found little support for the use of EQ assessment and only weak support for some distinct personality factors (ie, agreeableness, extraversion, and independence) in surgery resident selection. Performance on the SJT was associated with overall resident performance more than traditional cognitive measures (ie, USMLE scores). These data support further exploration of these 2 screening assessments on a larger scale across specialties and institutions.


[Lymphocyte coreceptors].
The activation of the immune system is tightly regulated by positive and negative receptors that allow the fine tuning of the immune cells. This regulation relies on receptors that were initially described in T lymphocytes, but have now been identified on cells from both innate and acquired immunity. The co-stimulatory receptors can allow cell activation or amplify it, regulate cell suvival and determine their effector functions. The co-inhibitory receptors can either prevent, decrease of inhibit the activation and differentiation process. The co-stimulatory and co-inhibitory molecules belong mainly to the so-called Ig superfamily and historically were called << CD28 and B7 family >>. The members of the tumor necrosis factor receptor (TNFR) family devoid of intra-cytoplasmic death domain but binding TNF receptor associated factors (TRAF) are also important but are up to now mainly co-stimulatory. The prototypical co-stimulatory molecules belonging to CD28 family are CD28 and ICOS, whereas the co-inhibitory molecules identified so far are CTLA-4, PD-1 and BTLA. Their receptors belong in most instances to the B7 family. For instance, B7.1/CD80 and B7.2/CD86 interact both with CD28 and CTLA-4 ; PDL1 and PDL2 bind to PD-1. The exception being so far BTLA which interacts with the TNFR family member HVEM (Herpes virus entry mediator). Three other B7 family members B7-H3, B7-H4 and BT3.1 are orphan receptors until now. The basis of co-inhibition rely on distinct mechanisms, one that has been postulated being the ability of the intracytoplasmic domain of PD-1 and BTLA to bind to the protein tyrosine phosphatases SHP-1 and SHP-2. The pathways used by the co-stimulatory receptors are also not completely understood and rely for CD28 both on signal similar to the one elicited by TcR and consequently increasing the overall signal and other more specific, elicited by the activation of PI3-OH kinase, vav1 and rearrangement of cytoskeleton. Recently, reverse signaling has been described for B7 family members which further increases the spectrum of functions elicited by these families. Co-stimulation and co-inhibition are among the most promising molecules and pathways to be targeted by mAbs, recombinant proteins and drugs in auto-immune diseases, transplantation and cancer.


Neutral fragment mass spectra via ambient thermal dissociation of peptide and protein ions.
A novel method for the fragmentation of peptide and protein ions at atmospheric pressure outside the mass spectrometer is described. Peptide/protein ions generated by electrosonic spray ionization (ESSI) are carried through a heated coiled metal tube where they fragment. Fragment ions of types a, b, and y are observed for peptides such as angiotensin II and bradykinin. In the case of phosphopeptides, informative b and y ions which preserve the labile phosphate groups are observed in the negative ion mode, which is potentially useful in the location of phosphorylation sites in proteins through chemical analysis of phosphopeptides. The thermal dissociation method extends to proteins such as ubiquitin and myoglobin, giving rise to y-type and other fragment ions. The most important feature of this method is that it also allows characterization of the neutral fragments arising from thermal dissociation by use of on-line corona discharge ionization. This neutral re-ionization experiment is much easier to perform outside the mass spectrometer than as conventionally done, in vacuum. It yields increased structural information from the resulting mass spectra in both the positive and the negative ion modes.


Emerging issues in paediatric health research consent forms in Canada: working towards best practices.
Obtaining a research participant's voluntary and informed consent is the bedrock of sound ethics practice. Greater inclusion of children in research has led to questions about how paediatric consent operates in practice to accord with current and emerging legal and socio-ethical issues, norms, and requirements. Employing a qualitative thematic content analysis, we examined paediatric consent forms from major academic centres and public organisations across Canada dated from 2008-2011, which were purposively selected to reflect different types of research ethics boards, participants, and studies. The studies included biobanking, longitudinal studies, and gene-environment studies. Our purpose was to explore the following six emerging issues: (1) whether the scope of parental consent allows for a child's assent, dissent, or future consent; (2) whether the concepts of risk and benefit incorporate the child's psychological and social perspective; (3) whether a child's ability to withdraw is respected and to what extent withdrawal is permitted; (4) whether the return of research results includes individual results and/or incidental findings and the processes involved therein; (5) whether privacy and confidentiality concerns adequately address the child's perspective and whether standard data and/or sample identifiability nomenclature is used; and (6) whether retention of and access to paediatric biological samples and associated medical data are addressed. The review suggests gaps and variability in the consent forms with respect to addressing each of the six issues. Many forms did not discuss the possibility of returning research results, be they individual or general/aggregate results. Forms were also divided in terms of the scope of parental consent (specific versus broad), and none discussed a process for resolving disputes that can arise when either the parents or the child wishes to withdraw from the study. The analysis provides valuable insight and evidence into how consent forms address current ethical issues. While we do not thoroughly explore the contexts and reasons behind consent form gaps and variability, we do advocate and formulate the development of best practices for drafting paediatric health research consent forms. This can greatly ameliorate current gaps and facilitate harmonised and yet contextualised approaches to paediatric health research ethics.


The use of segmented regression in analysing interrupted time series studies: an example in pre-hospital ambulance care.
An interrupted time series design is a powerful quasi-experimental approach for evaluating effects of interventions introduced at a specific point in time. To utilize the strength of this design, a modification to standard regression analysis, such as segmented regression, is required. In segmented regression analysis, the change in intercept and/or slope from pre- to post-intervention is estimated and used to test causal hypotheses about the intervention. We illustrate segmented regression using data from a previously published study that evaluated the effectiveness of a collaborative intervention to improve quality in pre-hospital ambulance care for acute myocardial infarction (AMI) and stroke. In the original analysis, a standard regression model was used with time as a continuous variable. We contrast the results from this standard regression analysis with those from segmented regression analysis. We discuss the limitations of the former and advantages of the latter, as well as the challenges of using segmented regression in analysing complex quality improvement interventions. Based on the estimated change in intercept and slope from pre- to post-intervention using segmented regression, we found insufficient evidence of a statistically significant effect on quality of care for stroke, although potential clinically important effects for AMI cannot be ruled out. Segmented regression analysis is the recommended approach for analysing data from an interrupted time series study. Several modifications to the basic segmented regression analysis approach are available to deal with challenges arising in the evaluation of complex quality improvement interventions.


Explosive genetic evidence for explosive human population growth.
The advent of next-generation sequencing technology has allowed the collection of vast amounts of genetic variation data. A recurring discovery from studying larger and larger samples of individuals had been the extreme, previously unexpected, excess of very rare genetic variants, which has been shown to be mostly due to the recent explosive growth of human populations. Here, we review recent literature that inferred recent changes in population size in different human populations and with different methodologies, with many pointing to recent explosive growth, especially in European populations for which more data has been available. We also review the state-of-the-art methods and software for the inference of historical population size changes that lead to these discoveries. Finally, we discuss the implications of recent population growth on personalized genomics, on purifying selection in the non-equilibrium state it entails and, as a consequence, on the genetic architecture underlying complex disease and the performance of mapping methods in discovering rare variants that contribute to complex disease risk.


Rearrangement phylogeny of genomes in contig form.
There has been a trend in increasing the phylogenetic scope of genome sequencing while decreasing the quality of the published sequence for each genome. With reduced finishing effort, there is an increasing number of genomes being published in contig form. Rearrangement algorithms, including gene order-based phylogenetic tools, require whole genome data on gene order, segment order, or some other marker order. Items whose chromosomal location is unknown cannot be part of the input. The question we address here is, for gene order-based phylogenetic analysis, how can we use rearrangement algorithms to handle genomes available in contig form only? Our suggestion is to use the contigs directly in the rearrangement algorithms as if they were chromosomes, while making a number of corrections, e.g., we correct for the number of extra fusion/fission operations required to make contigs comparable to full assemblies. We model the relationship between contig number and genomic distance, and estimate the parameters of this model using insect genome data. With this model, we use distance matrix methods to reconstruct the phylogeny based on genomic distance and numbers of contigs. We compare this with methods to reconstruct ancestral gene orders using uncorrected contig data.


The siderophore pyoverdine of Pseudomonas syringae pv. tabaci 6605 is an intrinsic virulence factor in host tobacco infection.
To investigate the role of iron uptake mediated by the siderophore pyoverdine in the virulence of the plant pathogen Pseudomonas syringae pv. tabaci 6605, three predicted pyoverdine synthesis-related genes, pvdJ, pvdL, and fpvA, were mutated. The pvdJ, pvdL, and fpvA genes encode the pyoverdine side chain peptide synthetase III L-Thr-L-Ser component, the pyoverdine chromophore synthetase, and the TonB-dependent ferripyoverdine receptor, respectively. The Delta pvdJ and Delta pvdL mutants were unable to produce pyoverdine in mineral salts-glucose medium, which was used for the iron-depleted condition. Furthermore, the Delta pvdJ and Delta pvdL mutants showed lower abilities to produce tabtoxin, extracellular polysaccharide, and acyl homoserine lactones (AHLs), which are quorum-sensing molecules, and consequently had reduced virulence on host tobacco plants. In contrast, all of the mutants had accelerated swarming ability and increased biosurfactant production, suggesting that swarming motility and biosurfactant production might be negatively controlled by pyoverdine. Scanning electron micrographs of the surfaces of tobacco leaves inoculated with the mutant strains revealed only small amounts of extracellular polymeric matrix around these mutants, indicating disruption of the mature biofilm. Tolerance to antibiotics was drastically increased for the Delta pvdL mutant, as for the Delta psyI mutant, which is defective in AHL production. These results demonstrated that pyoverdine synthesis and the quorum-sensing system of Pseudomonas syringae pv. tabaci 6605 are indispensable for virulence in host tobacco infection and that AHL may negatively regulate tolerance to antibiotics.