The characteristics of a good clinical teacher as perceived by resident physicians in Japan: a qualitative study.
It is not known whether the characteristics of a good clinical teacher as perceived by resident physicians are the same in Western countries as in non-Western countries including Japan. The objective of this study was to identify the characteristics of a good clinical teacher as perceived by resident physicians in Japan, a non-Western country, and to compare the results with those obtained in Western countries. Data for this qualitative research were collected using semi-structured focus group interviews. Focus group transcripts were independently analyzed and coded by three authors. Residents were recruited by maximum variation sampling until thematic saturation was achieved. Twenty-three residents participated in five focus group interviews regarding the perceived characteristics of a good clinical teacher in Japan. The 197 descriptions of characteristics that were identified were grouped into 30 themes. The most commonly identified theme was "provided sufficient support", followed by "presented residents with chances to think", "provided feedback", and "provided specific indications of areas needing improvement". Using Sutkin's main categories (teacher, physician, and human characteristics), 24 of the 30 themes were categorized as teacher characteristics, 6 as physician characteristics, and none as human characteristics. "Medical knowledge" of teachers was not identified as a concern of residents, and "clinical competence of teachers" was not emphasized, whereas these were the two most commonly recorded themes in Sutkin's study. Our results suggest that Japanese and Western resident physicians place emphasis on different characteristics of their teachers. We speculate that such perceptions are influenced by educational systems, educational settings, and culture. Globalization of medical education is important, but it is also important to consider differences in educational systems, local settings, and culture when evaluating clinical teachers.


Role of platelet transfusion in the management of dengue patients in a tertiary care hospital.
While medical fraternity globally recognizes the role of platelet transfusion in the management of hospitalized dengue patients the exact indications and situations in which these are to be transfused may vary. Since there is inherent risk associated with the transfusion of blood/blood-component, it is imperative for each institution (or country) to lay their own criteria for transfusion of these blood components. The present study was conducted to lay precise criteria and transfusion trigger for platelet transfusion in our set-up. The present study was conducted on 225 serologically confirmed dengue patients admitted at Indraprastha Apollo Hospitals between 1(st) of August to 30(th) of November 2005. Clinical data, reports of hematological investigation, platelets requirements and data obtained from daily follow-up were analyzed. The clinicians followed the guidelines issued by the Directorate of Health services, NCT of Delhi. In the serologically confirmed cases, the prevalence of thrombocytopenia (count less than 100,000/cumm) was 84.88% on admission and bleeding was recorded in 22 (9.7%) patients. About 96 (42.6%) patients of dengue cases received platelet transfusion. Among them 47 (20.88%) patients had a platelet count < 20,000/cumm, 43 (19.11%) had a platelet count in the range of 21-40,000/cumm while 6 (2.66%) patients had the platelet count in between 41 and 50,000/cumm. Out of 49 patients with a platelet count >20,000/cumm, 18 patients had haemorrhagic manifestations such as petechiae, gum-bleeding, epistaxis, etc., which necessitates the use of platelet transfusion. However, 31 patients received inappropriate platelet transfusion. This study suggests that bleeding occurs more often in patients with severe thrombocytopenia. High-risk patients having platelet count < 20,000/cumm and risk of bleeding require urgent platelet transfusion. Patients with platelet count 21-40,000/cumm are in moderate risk and require platelet transfusion only if they have any haemorrhagic manifestations and other superadded conditions.


Use of a new RNA next generation sequencing approach for the specific detection of virus infection in cells.
The utilization of the current combination of in vitro, in vivo and PCR assays for the identification of adventitious viruses in production cells has a limited range of detection. While Next Generation Sequencing (NGS) has a broader breadth of detection, it is unable to differentiate sequences from replicating viruses versus background inert sequences. In order to improve NGS specificity, we have designed a new NGS approach which targets subsets of viral RNAs only synthesized during cell infection. In order to evaluate the performance of this approach for detecting low levels of adventitious viruses, we selected two difficult virus/cell systems. This included B95-8 cells persistently infected by Human herpesvirus 4 (HHV-4) and serially diluted into HHV-4 negative Ramos cells and Madin-Darby bovine kidney cells with an early infection produced via a low dose of Bovine viral diarrhea virus. We demonstrated that the sensitivity of our RNA NGS approach was equivalent to targeted PCR with an increased specificity for the detection of viral infection. We were also able to identify a previously undetected Murine Leukemia Virus contaminant in Ramos cells. Based on these results, we conclude that this new RNA NGS approach is suitable for conducting viral safety evaluations of cells.


Rhodopsin mutants that bind but fail to activate transducin.
Rhodopsin is a member of a family of receptors that contain seven transmembrane helices and are coupled to G proteins. The nature of the interactions between rhodopsin mutants and the G protein, transduction (Gt), was investigated by flash photolysis in order to monitor directly Gt binding and dissociation. Three mutant opsins with alterations in their cytoplasmic loops bound 11-cis-retinal to yield pigments with native rhodopsin absorption spectra, but they failed to stimulate the guanosine triphosphatase activity of Gt. The opsin mutations included reversal of a charged pair conserved in all G protein-coupled receptors at the cytoplasmic border of the third transmembrane helix (mutant CD1), replacement of 13 amino acids in the second cytoplasmic loop (mutant CD2), and deletion of 13 amino acids from the third cytoplasmic loop (mutant EF1). Whereas mutant CD1 failed to bind Gt, mutants CD2 and EF1 showed normal Gt binding but failed to release Gt in the presence of guanosine triphosphate. Therefore, it appears that at least the second and third cytoplasmic loops of rhodopsin are required for activation of bound Gt.


Palliative care in cardiopulmonary transplantation.
Cardiopulmonary transplantation is a life-prolonging therapy available to a select population of patients with cardiac or respiratory failure. Transplantation is associated with significant morbidity, mortality and unmet palliative care need. Despite recommendations that palliative care should be a core component of the heart and lung transplant process, collaboration within clinical practice is extremely rare. A key reason for this is the misperception among patients, their families and transplant clinicians, that palliative care is analogous with end of life care. Other challenges include prognostication, communication, and the balance of hope and reality. We suggest a change in clinical practice within cardiopulmonary transplantation, whereby palliative care takes place alongside active management. Greater partnership working will demonstrate clinical credibility and highlight the impact of palliative care interventions. Education is required to address current misperceptions and further research should explore the effect of initiatives to improve palliative care provision for this patient group.


[Comparison of 5 methods to estimate the prevalence of diabetes mellitus in population-based studies].
To aid in the search for more practical and reliable methods for use in population-based studies of diabetes mellitus, this article compares five ways of estimating prevalence rates. The analysis was performed on secondary data from a cross-sectional study of a cluster sample of the adult population in nine state capitals in Brazil. The original study was carried out from 1986 to 1988. The 21,846 participants were classified as diabetic or not diabetic by five different methods: household questionnaires administered to the entire sample population (M1); individual questionnaires administered to the selected population (M2); measurement of fasting glucose levels in capillary blood, with levels > or = 120 mg/dL as the cutoff (M3); individual questionnaire and fasting capillary blood glucose > or = 120 mg/dL (M4); and individual questionnaire plus fasting capillary blood glucose > or = 120 mg/dL and capillary glucose 2 hours after oral glucose loading > or = 200 mg/dL (M5). Agreement between the methods was determined by comparison of the rates obtained and use of the kappa coefficient. The age-adjusted prevalence rates of diabetes varied according to the method used. Values obtained with M1 were lower than those indicated by M2; M3 values were higher than M2 values, except in the age group 60-69 years; and with M5 the rates were higher than with M4, except among persons 30-39 years old. With regard to the age-adjusted rates found by the various methods, M1 detected 84% of the M2 estimate, M2 detected 91% of the M3, M3 detected 70% of the M4, and M4 detected 86% of the M5. Previously diagnosed diabetes cases accounted for 64% and 55% of the totals estimated by M4 and M5, respectively. Kappa values were at least 0.70 for M1 compared against M2, M1 against M4, M2 against M4, and M3 against M4. Based on the results of this study, it was concluded that the questionnaires used in M1 and M2 constituted appropriate methods for detecting previously diagnosed cases of diabetes mellitus, and their use is recommended for the purposes of health services planning or evaluation. Fasting glucose measurement (M3) as the sole method did not show a significant advantage over the individual questionnaire (M2). Of the combined or multiple methods, fasting glucose together with the individual questionnaire (M4) was efficient in comparison to M5, which incorporated measurement of blood glucose 2 hours after oral glucose ingestion.


An evaluation of space time cube representation of spatiotemporal patterns.
Space time cube representation is an information visualization technique where spatiotemporal data points are mapped into a cube. Information visualization researchers have previously argued that space time cube representation is beneficial in revealing complex spatiotemporal patterns in a data set to users. The argument is based on the fact that both time and spatial information are displayed simultaneously to users, an effect difficult to achieve in other representations. However, to our knowledge the actual usefulness of space time cube representation in conveying complex spatiotemporal patterns to users has not been empirically validated. To fill this gap, we report on a between-subjects experiment comparing novice users' error rates and response times when answering a set of questions using either space time cube or a baseline 2D representation. For some simple questions, the error rates were lower when using the baseline representation. For complex questions where the participants needed an overall understanding of the spatiotemporal structure of the data set, the space time cube representation resulted in on average twice as fast response times with no difference in error rates compared to the baseline. These results provide an empirical foundation for the hypothesis that space time cube representation benefits users analyzing complex spatiotemporal patterns.


Common mental disorders and long-term sickness absence in a general working population. The Hordaland Health Study.
To examine and compare the prospective effect of the common mental disorders (CMD) anxiety and depression on duration and recurrence of sickness absence (SA), and to investigate whether the effect of CMD on SA is detectable over time. Information from a large epidemiological health study (N = 13 436) was linked with official records of SA episodes lasting ≥16 days up to 6 years after participation. Common mental disorders were assessed with the Hospital Anxiety and Depression Scale (HADS). Associations were analysed with Cox regression and multinomial logistic regression models controlling for potential covariates. Comorbid anxiety and depression, and anxiety only were significant risk factors for SA after adjusting for covariates, whilst depression only was not. Anxiety and depression were stronger predictors for longer duration of SA episodes compared with shorter duration and associated with more frequent recurrence of SA. There was a general trend toward the effect of CMD on SA becoming weaker over time; however, the effect of anxiety only on SA remained stable throughout the follow-up. Common mental disorders are long-lasting predictors of onset, duration and recurrence of SA. Anxiety appears to be a more important contributor to long-term SA than previously described in the literature.


Corneal biomechanical properties in primary open angle glaucoma and normal tension glaucoma.
To determine and compare the corneal biomechanical properties between eyes with primary open angle glaucoma (POAG) and eyes with normal tension glaucoma (NTG). Prospective cross-sectional study. Consecutive eligible POAG and NTG patients attending the Glaucoma Clinic had assessment of their corneal biomechanical properties-corneal hysteresis (CH) and corneal resistance factor (CRF)-using the Ocular Response Analyzer by an observer masked to the diagnosis. Exclusion criteria included previous intraocular surgery, corneal pathology, inflammatory connective tissue disease, and refraction of 5-dimensional or over. If both eyes were eligible, then the right eye was used for analysis. The main outcome measures were corneal hysteresis and CRF measurements. Data analysis was performed using the t test and general linear model. Eighty-one patients (80 whites) were analyzed. Forty had NTG, whereas 41 had POAG. Thirty-five were females. There was a statistically significant difference in mean CH (NTG 9.6+/-1.3 mm Hg; POAG 9.0+/-1.4 mm Hg; P=0.01), but not in mean CRF (NTG 9.9+/-1.4; POAG 10.8+/-1.7; P=0.06). The highest recorded Goldmann applanation intraocular pressure (IOP) was statistically significantly associated with lower CH (P=0.01) and higher CRF (P=0.02). There was a small but statistically significant difference in the mean CH between POAG and NTG (CH was higher in NTG). The highest recorded Goldmann applanation IOP was also statistically significantly correlated with lower CH and higher CRF, suggesting that alterations to the corneal biomechanical properties may occur as a result of chronic raised IOP in POAG.


Guideline update for the performance of fusion procedures for degenerative disease of the lumbar spine. Part 10: lumbar fusion for stenosis without spondylolisthesis.
Lumbar stenosis is one of the more common radiographic manifestations of the aging process, leading to narrowing of the spinal canal and foramen. When stenosis is clinically relevant, patients often describe activity-related low-back or lower-extremity pain, known as neurogenic claudication. For those patients who do not improve with conservative care, surgery is considered an appropriate treatment alternative. The primary objective of surgery is to reconstitute the spinal canal. The role of fusion, in the absence of a degenerative deformity, is uncertain. The previous guideline recommended against the inclusion of lumbar fusion in the absence of spinal instability or a likelihood of iatrogenic instability. Since the publication of the original guidelines, numerous studies have demonstrated the role of surgical decompression in this patient population; however, few have investigated the utility of fusion in patients without underlying instability. The majority of studies contain a heterogeneous cohort of subjects, often combining patients with and without spondylolisthesis who received various surgical interventions, limiting fusions to those patients with instability. It is difficult if not impossible, therefore, to formulate valid conclusions regarding the utility of fusion for patients with uncomplicated stenosis. Lower-level evidence exists, however, that does not demonstrate an added benefit of fusion for these patients; therefore, in the absence of deformity or instability, the inclusion of a fusion is not recommended.


Cancer and Leukemia Group B 90206: A randomized phase III trial of interferon-alpha or interferon-alpha plus anti-vascular endothelial growth factor antibody (bevacizumab) in metastatic renal cell carcinoma.
The majority of sporadic clear cell renal cell carcinoma (RCC) is characterized by loss of heterozygosity of the von Hippel-Lindau (VHL) tumor suppressor gene and somatic inactivation of the remaining VHL allele. The resulting VHL gene silencing leads to induction of hypoxia-regulated genes including vascular endothelial growth factor (VEGF). Thus, therapeutic inhibition of VEGF holds promise for treatment of this historically refractory malignancy. An antibody to VEGF (bevacizumab, Avastin) has demonstrated a significant prolongation of time to disease progression compared with placebo in patients with metastatic RCC. Interferon-alpha (IFN-alpha) is a standard initial cytokine therapy in RCC with a modest response rate and a survival advantage demonstrated in randomized trials. We hypothesized that the addition of anti-VEGF therapy to IFN-alpha would prolong survival in untreated metastatic RCC patients. A Phase III trial is now being conducted randomizing untreated, metastatic clear cell RCC patients to IFN-alpha alone or IFN-alpha plus Avastin.


Energy requirements of adults.
To describe issues related to energy requirements of free living adults and discuss the importance of basal metabolic rate (BMR) and their relationships to total energy expenditure (TEE ) and physical activity level (PAL, derived as TEE/BMR) and to determine the influence of body weight, height, age and sex. Based on a review of the literature, this paper examines the variability in BMR due to methodology, ethnicity, migration and adaptation (both metabolic and behavioural) due to changes in nutritional status. Collates and compiles data on measurements of TEE in free living healthy adults, to arrive at limits and to compare TEE of populations with different life-styles. The constancy of BMR and its validity as a reliable predictor of TEE in adults as well as the validity of PAL as an index of TEE adjusted for BMR and thus its use to categorise the physical activity pattern and lifestyle of an individual was confirmed. The limits of human daily energy expenditure at around 1.2 x BMR and 4.5 x BMR based on measurements made in free living adults have been reported in the literature. A large and robust database now exists of energy expenditure measurements obtained by the doubly labelled water method in the scientific literature and the data shows that, in general, levels of energy expenditure are similar to the recommendations for energy requirements adopted by FAO/WHO/UNU (1985). The review also confirms that metabolic adaptation to energy restriction is not an important factor that needs to be considered when recommending energy requirements for adults in developing countries.


The reliability of tweets as a supplementary method of seasonal influenza surveillance.
Existing influenza surveillance in the United States is focused on the collection of data from sentinel physicians and hospitals; however, the compilation and distribution of reports are usually delayed by up to 2 weeks. With the popularity of social media growing, the Internet is a source for syndromic surveillance due to the availability of large amounts of data. In this study, tweets, or posts of 140 characters or less, from the website Twitter were collected and analyzed for their potential as surveillance for seasonal influenza. There were three aims: (1) to improve the correlation of tweets to sentinel-provided influenza-like illness (ILI) rates by city through filtering and a machine-learning classifier, (2) to observe correlations of tweets for emergency department ILI rates by city, and (3) to explore correlations for tweets to laboratory-confirmed influenza cases in San Diego. Tweets containing the keyword "flu" were collected within a 17-mile radius from 11 US cities selected for population and availability of ILI data. At the end of the collection period, 159,802 tweets were used for correlation analyses with sentinel-provided ILI and emergency department ILI rates as reported by the corresponding city or county health department. Two separate methods were used to observe correlations between tweets and ILI rates: filtering the tweets by type (non-retweets, retweets, tweets with a URL, tweets without a URL), and the use of a machine-learning classifier that determined whether a tweet was "valid", or from a user who was likely ill with the flu. Correlations varied by city but general trends were observed. Non-retweets and tweets without a URL had higher and more significant (P<.05) correlations than retweets and tweets with a URL. Correlations of tweets to emergency department ILI rates were higher than the correlations observed for sentinel-provided ILI for most of the cities. The machine-learning classifier yielded the highest correlations for many of the cities when using the sentinel-provided or emergency department ILI as well as the number of laboratory-confirmed influenza cases in San Diego. High correlation values (r=.93) with significance at P<.001 were observed for laboratory-confirmed influenza cases for most categories and tweets determined to be valid by the classifier. Compared to tweet analyses in the previous influenza season, this study demonstrated increased accuracy in using Twitter as a supplementary surveillance tool for influenza as better filtering and classification methods yielded higher correlations for the 2013-2014 influenza season than those found for tweets in the previous influenza season, where emergency department ILI rates were better correlated to tweets than sentinel-provided ILI rates. Further investigations in the field would require expansion with regard to the location that the tweets are collected from, as well as the availability of more ILI data.


Pneumococcal vaccine impacts on the population genomics of non-typeable Haemophilus influenzae.
The implementation of pneumococcal conjugate vaccines (PCVs) has led to a decline in vaccine-type disease. However, there is evidence that the epidemiology of non-typeable Haemophilus influenzae (NTHi) carriage and disease can be altered as a consequence of PCV introduction. We explored the epidemiological shifts in NTHi carriage using whole genome sequencing over a 5-year period that included PCV13 replacement of PCV7 in the UK's National Immunization Programme in 2010. Between 2008/09 and 2012/13 (October to March), nasopharyngeal swabs were taken from children <5 years of age. Significantly increased carriage post-PCV13 was observed and lineage-specific associations with Streptococcus pneumoniae were seen before but not after PCV13 introduction. NTHi were characterized into 11 discrete, temporally stable lineages, congruent with current knowledge regarding the clonality of NTHi. The increased carriage could not be linked to the expansion of a particular clone and different co-carriage dynamics were seen before PCV13 implementation when NTHi co-carried with vaccine serotype pneumococci. In summary, PCV13 introduction has been shown to have an indirect effect on NTHi epidemiology and there exists both negative and positive, distinct associations between pneumococci and NTHi. This should be considered when evaluating the impacts of pneumococcal vaccine design and policy.


miR-16 and miR-21 expression in the placenta is associated with fetal growth.
Novel research has suggested that altered miRNA expression in the placenta is associated with adverse pregnancy outcomes and with potentially harmful xenobiotic exposures. We hypothesized that aberrant expression of miRNA in the placenta is associated with fetal growth, a measurable phenotype resulting from a number of intrauterine factors, and one which is significantly predictive of later life outcomes. We analyzed 107 primary, term, human placentas for expression of 6 miRNA reported to be expressed in the placenta and to regulate cell growth and development pathways: miR-16, miR-21, miR-93, miR-135b, miR-146a, and miR-182. The expression of miR-16 and miR-21 was markedly reduced in infants with the lowest birthweights (p<0.05). Logistic regression models suggested that low expression of miR-16 in the placenta predicts an over 4-fold increased odds of small for gestational age (SGA) status (p = 0.009, 95% CI = 1.42, 12.05). Moreover, having both low miR-16 and low miR-21 expression in the placenta predicts a greater increase in odds for SGA than having just low miR-16 or miR-21 expression (p<0.02), suggesting an additive effect of both of these miRNA. Our study is one of the first to investigate placental miRNA expression profiles associated with birthweight and SGA status. Future research on miRNA whose expression is associated with in utero exposures and markers of fetal growth is essential for better understanding the epigenetic mechanisms underlying the developmental origins of health and disease.


Computational approach to quantifying hemodynamic forces in giant cerebral aneurysms.
The options for treating giant fusiform basilar aneurysms are limited, and the potential impact of planned interventions is difficult to assess. We developed a computational framework to evaluate the impact that interventions might have on hemodynamic conditions. A computational fluid dynamics approach was used to determine the velocity field, wall shear stress, and pressure distribution within a model of a basilar artery before and after a simulated occlusion of one vertebral artery. The vascular geometry in a patient with a giant fusiform basilar artery aneurysm was determined by using contrast-enhanced MR angiography, and the numerical simulation approach was used to calculate the flow fields in the presenting geometry and to predict the flow field that would occur if a vertebral artery were occluded. In the model geometry, computational fluid dynamics indicated that there would be a symmetric flow pattern with a strong central stream and large recirculation zones at the walls. After simulated occlusion of one vertebral artery, the primary stream was diverted to one side, resulting in high pressure and increased wall shear stress. For the patient-specific geometry, flow patterns were shown to depend strongly on how much flow there was in each vertebral artery. Contrast-enhanced MR angiography is an effective tool for demonstrating the luminal boundaries of large intracranial aneurysms. Computational fluid dynamics is a powerful tool for determining the prevailing flow conditions in vascular territories and for modeling the possible alterations of the flow field that would result from interventional treatments.


Evaluating complex healthcare systems: a critique of four approaches.
The purpose of this paper is to bring clarity to the emerging conceptual and methodological literature that focuses on understanding and evaluating complex or 'whole' systems of healthcare. An international working group reviewed literature from interdisciplinary or interprofessional groups describing approaches to the evaluation of complex systems of healthcare. The following four key approaches were identified: a framework from the MRC (UK), whole systems research, whole medical systems research described by NCCAM (USA) and a model from NAFKAM (Norway). Main areas of congruence include acknowledgment of the inherent complexity of many healthcare interventions and the need to find new ways to evaluate these; the need to describe and understand the components of complex interventions in context (as they are actually practiced); the necessity of using mixed methods including randomized clinical trials (RCTs) (explanatory and pragmatic) and qualitative approaches; the perceived benefits of a multidisciplinary team approach to research; and the understanding that methodological developments in this field can be applied to both complementary and alternative medicine (CAM) as well as conventional therapies. In contrast, the approaches differ in the following ways: terminology used, the extent to which the approach attempts to be applicable to both CAM and conventional medical interventions; the prioritization of research questions (in order of what should be done first) especially with respect to how the 'definitive' RCT fits into the process of assessing complex healthcare systems; and the need for a staged approach. There appears to be a growing international understanding of the need for a new perspective on assessing complex healthcare systems.


Social norm influences on evaluations of the risks associated with alcohol consumption: applying the rank-based decision by sampling model to health judgments.
The research first tested whether perceptions of other people's alcohol consumption influenced drinkers' perceptions of the riskiness of their own consumption. Second, the research tested how such comparisons are made-whether, for example, people compare their drinking to the 'average' drinker's or 'rank' their consumption amongst other people's. The latter untested possibility, suggested by the recent Decision by Sampling Model of judgment, would imply different cognitive mechanisms and suggest that information should be presented differently to people in social norm interventions. Study 1 surveyed students who provided information on (a) their own drinking, (b) their perceptions of the distribution of drinking in the UK and (c) their perceived risk of various alcohol-related disorders. Study 2 experimentally manipulated the rank of 'target' units of alcohol within the context of units viewed simultaneously. In both studies, the rank of an individual's drinking in a context of other drinkers predicted perceptions of developing alcohol-related disorders. There was no evidence for the alternative hypothesis that people compared with the average of other drinkers' consumptions. The position that subjects believed they occupied in the ranking of other drinkers predicted their perceived risk, and did so as strongly as how much they actually drank. Drinking comparisons are rank-based, which is consistent with other judgments in social, emotional and psychophysical domains. Interventions should be designed to work with people's natural ways of information processing, through providing clients with information on their drinking rank rather than how their drinking differs from the average.


On the complexity of shark bite wounds: From associated bacteria to trauma management and wound repair.
The mouth of a shark is a breeding ground for a large variety of bacteria which can easily get transferred onto a human body in the event of a shark bite. Here, we review infections originating from shark oral bacterial flora, which originate from the microbiome of its prey, as well as from the surroundings where an incident takes place. We use the example of an incident which occurred in an aquarium involving a sandtiger shark, Carcharias taurus. In addition to a succinct analysis and interpretation of the wound and recovery process, an overview of currently known bacteria associated with shark bite wounds is given, as well as a summary of the effects of various previously tested antibiotics on bacteria derived from blacktip sharks, Carcharhinus limbatus, bull sharks, C. leucas, and tiger sharks, Galeocerdo cuvier. Therapeutic study, level V.


Thrombolysis of the cervical internal carotid artery before balloon angioplasty and stent placement: report of two cases.
The application of endovascular techniques to the treatment of cervical carotid artery bifurcation atherosclerosis has been delayed because of the fear of causing embolic events while traversing the diseased portion of the artery with an angioplasty balloon catheter. Symptomatic carotid arteries often contain fresh or partially digested intraluminal thrombus. Before we cross certain carotid bifurcation lesions with angioplasty catheters, we deliver 100,000 to 200,000 units of urokinase in an attempt to digest loose thrombus. We have witnessed changes in the angiographic appearance of the diseased portion of the vessel after urokinase treatment, such as widening of the lumen, that suggest clot lysis. We present two patients who had symptomatic internal carotid artery stenosis. Angiography showed irregular narrowing of the internal carotid artery origin. One patient was selected for angioplasty instead of carotid endarterectomy because of severe cardiac risk factors. The other patient had major angiographic risk factors manifested by poor collateral circulation. The angiographic findings and history of transient ischemic attacks led us to suspect the presence of soft, loose plaque debris or thrombus in both cases. Therefore, we performed thrombolysis with urokinase before angioplasty. Repeat angiography showed widening of the arterial lumen and smoothing of the plaque profile. Subsequent angioplasty and stent placement were uneventful. Intraarterial thrombolysis can produce a change in the angiographic appearance of symptomatic atherosclerotic lesions of the cervical carotid artery bifurcation. Digestion of intralesional thrombus may provide a safer environment for deployment of endovascular remodeling devices by decreasing the likelihood of embolic phenomena. We believe thrombolysis should be done before angioplasty in select patients.


The products of the SPT10 and SPT21 genes of Saccharomyces cerevisiae increase the amplitude of transcriptional regulation at a large number of unlinked loci.
The 3' long terminal repeat (LTR) of yeast transposon Ty1 is not normally used as a promoter, although it contains sequences identical to those found in the 5' LTR, which does act as a promoter. We have isolated mutations that fall into two genes, SPT10 and SPT21, that allow the 3' LTRs of Ty1 elements inserted at various positions in the genome of Saccharomyces cerevisiae to act as promoters. We find that mutations in these two genes alter transcriptional regulation of Ty1 LTRs and also of certain non-Ty1-related promoters in two ways: (i) they allow the low-level expression of several genes under repressing conditions, and (ii) they allow transcription from the 5' LTR of Ty1 elements in the absence of a normally required activator, SPT3. Furthermore, the fully induced levels of transcription of several genes are reduced in these spt mutants. Hence, the products of these two genes increase the amplitude of transcriptional regulation of a wide variety of unlinked loci.


Excessive postrotary nystagmus duration in learning-disabled children.
The Southern California Sensory Integration Test results of 109 learning-disabled children were reviewed and analyzed to investigate the hypothesis that learning-disabled children with excessive postrotary nystagmus durations display greater neuropsychological impairment than learning-disabled children with normal or depressed postrotary nystagmus. The findings supported the hypothesis that learning-disabled children judged to have relatively greater neuropsychological involvement exhibited significantly longer postrotary nystagmus durations. Data analysis revealed that six Southern California Sensory Integration Tests assumed to measure cortical level functions shared significant variance with excessive Southern California Postrotary Nystagmus Test (SCPNT) scores. A similar relationship was not found for normal or depressed SCPNT scores. A connection between cortical level versus brain-stem level central nervous system dysfunction, nystagmus, and learning disorder is discussed.


Adaptive seasonal shift towards investment in fewer, larger offspring: Evidence from field and laboratory studies.
Seasonal changes in reproduction have been described for many taxa. As reproductive seasons progress, females often shift from greater energetic investment in many small offspring towards investing less total energy into fewer, better provisioned (i.e. larger) offspring. The underlying causes of this pattern have not been assessed in many systems. Two primary hypotheses have been proposed to explain these patterns. The first is an adaptive hypothesis from life-history theory: early offspring have a survival advantage over those produced later. Accordingly, selection favours females that invest in offspring quantity early in the season and offspring quality later. The second hypothesis suggests these patterns are not intrinsic but result from passive responses to seasonal changes in the environment experienced by reproducing females (i.e. maternal environment). To disentangle the causes underlying this pattern, which has been reported in brown anole lizards (Anolis sagrei), we performed complementary field and laboratory studies. The laboratory study carefully controlled maternal environments and quantified reproductive patterns throughout the reproductive season for each female. The field study measured similar metrics from free ranging lizards across an entire reproductive season. In the laboratory, females increased relative effort per offspring as the reproductive season progressed; smaller eggs were laid earlier, larger eggs were laid later. Moreover, we observed significant among-individual variation in seasonal changes in reproduction, which is necessary for traits to evolve via natural selection. Because these patterns consistently emerge under controlled laboratory conditions, they likely represent an intrinsic and potentially adaptive adjustment of reproductive effort as predicted by life-history theory. The field study revealed similar trends, further suggesting that intrinsic patterns observed in the laboratory are strong enough to persist despite the environmental variability that characterizes natural habitats. The observed patterns are indicative of an adaptive seasonal shift in parental investment in response to a deteriorating offspring environment: allocating greater resources to late-produced offspring likely enhances maternal fitness.


Integrative and comparative genomic analysis of lung squamous cell carcinomas in East Asian patients.
Lung squamous cell carcinoma (SCC) is the second most prevalent type of lung cancer. Currently, no targeted therapeutics are approved for treatment of this cancer, largely because of a lack of systematic understanding of the molecular pathogenesis of the disease. To identify therapeutic targets and perform comparative analyses of lung SCC, we probed somatic genome alterations of lung SCC by using samples from Korean patients. We performed whole-exome sequencing of DNA from 104 lung SCC samples from Korean patients and matched normal DNA. In addition, copy-number analysis and transcriptome analysis were conducted for a subset of these samples. Clinical association with cancer-specific somatic alterations was investigated. This cancer cohort is characterized by a high mutational burden with an average of 261 somatic exonic mutations per tumor and a mutational spectrum showing a signature of exposure to cigarette smoke. Seven genes demonstrated statistical enrichment for mutation: TP53, RB1, PTEN, NFE2L2, KEAP1, MLL2, and PIK3CA). Comparative analysis between Korean and North American lung SCC samples demonstrated a similar spectrum of alterations in these two populations in contrast to the differences seen in lung adenocarcinoma. We also uncovered recurrent occurrence of therapeutically actionable FGFR3-TACC3 fusion in lung SCC. These findings provide new steps toward the identification of genomic target candidates for precision medicine in lung SCC, a disease with significant unmet medical needs.


Measuring what people value: a comparison of "attitude" and "preference" surveys.
To compare and contrast methods and findings from two approaches to valuation used in the same survey: measurement of "attitudes" using simple rankings and ratings versus measurement of "preferences" using conjoint analysis. Conjoint analysis, a stated preference method, involves comparing scenarios composed of attribute descriptions by ranking, rating, or choosing scenarios. We explore possible explanations for our findings using focus groups conducted after the quantitative survey. A self-administered survey, measuring attitudes and preferences for HIV tests, was conducted at HIV testing sites in San Francisco in 1999-2000 (n = 365, response rate = 96 percent). Attitudes were measured and analyzed using standard approaches. Conjoint analysis scenarios were developed using a fractional factorial design and results analyzed using random effects probit models. We examined how the results using the two approaches were both similar and different. We found that "attitudes" and "preferences" were generally consistent, but there were some important differences. Although rankings based on the attitude and conjoint analysis surveys were similar, closer examination revealed important differences in how respondents valued price and attributes with "halo" effects, variation in how attribute levels were valued, and apparent differences in decision-making processes. To our knowledge, this is the first study to compare attitude surveys and conjoint analysis surveys and to explore the meaning of the results using post-hoc focus groups. Although the overall findings for attitudes and preferences were similar, the two approaches resulted in some different conclusions. Health researchers should consider the advantages and limitations of both methods when determining how to measure what people value.


Comparative phylogeography of two widespread magpies: importance of habitat preference and breeding behavior on genetic structure in China.
Historical geological events and climatic changes are believed to have played important roles in shaping the current distribution of species. However, sympatric species may have responded in different ways to such climatic fluctuations. Here we compared genetic structures of two corvid species, the Azure-winged Magpie Cyanopica cyanus and the Eurasian Magpie Pica pica, both widespread but with different habitat dependence and some aspects of breeding behavior. Three mitochondrial genes and two nuclear introns were used to examine their co-distributed populations in East China and the Iberian Peninsula. Both species showed deep divergences between these two regions that were dated to the late Pliocene/early Pleistocene. In the East Chinese clade of C. cyanus, populations were subdivided between Northeast China and Central China, probably since the early to mid-Pleistocene, and the Central subclade showed a significant pattern of isolation by distance. In contrast, no genetic structure was found in the East China populations of P. pica. We suggest that the different patterns in the two species are at least partly explained by ecological differences between them, especially in habitat preference and perhaps also breeding behavior. These dissimilarities in life history traits might have affected the dispersal and survival abilities of these two species differently during environmental fluctuations.


Standardization of the preanalytical phase of DNA extraction from fixed tissue for next-generation sequencing analyses.
Next-generation sequencing (NGS) analyses on DNA derived from archived Formalin-Fixed Paraffin-Embedded (FFPE) clinical material can provide a powerful tool in oncology research and clinical diagnostics. Although several studies have established that NGS can be performed using DNA from FFPE tissue, the accuracy and reproducibility of such analyses, as well as their robustness to the biomolecular quality of the samples used, remains a matter of debate. Excellent reviews have recently been published, providing evidence-based best practices for FFPE DNA extraction. Alternative fixatives exist, although their implementation in clinical practice is difficult. In this article, we present (i) a review of fixed tissue DNA preanalytics with a special focus on DNA extraction and fixed tissue sample qualification and (ii) results from comparisons between different methods of DNA extraction from tissue samples that have been fixed or stabilized by different methods, in terms of NGS metrics and different DNA quality metrics.


Sex differences in clinical characteristics and outcomes after myocardial infarction: insights from the Valsartan in Acute Myocardial Infarction Trial (VALIANT).
We examined the association of sex with clinical characteristics and outcomes in patients following myocardial infarction (MI) in the Valsartan in Acute Myocardial Infarction Trial (VALIANT). A total of 4570 women and 10 133 men with heart failure (HF), left ventricular systolic dysfunction or both were enrolled 0.5-10 days after myocardial infarction (MI) and followed for a median of 24.7 months. Compared with men, women were older, had more comorbidities, and were more likely to present in Killip Class III/IV and experience post-infarction angina or HF. After adjusting for baseline differences, both short-term and longer-term mortality were similar in men and women. However, women were more likely than men to experience the composite outcome of cardiovascular death, MI, HF, stroke, and resuscitation from cardiac arrest [adjusted hazard ratio (HR) 1.15; 95% confidence interval (CI); 1.06-1.24, P=0.001], mainly owing to an increased risk of hospitalization for HF (adjusted HR 1.36; 95% CI 1.22-1.52; P<0.001). In a subset of patients who underwent echocardiographic study (n=603), women had smaller left ventricular volumes indexed by body size than men but similar ejection fractions and similar changes in ventricular volumes from baseline to 1 month and 20 months. In VALIANT, the risk of HF following MI was higher in women than men after adjusting for age and comorbidities, although the risk of other fatal and non-fatal outcomes were similar. The higher long-term risk of HF in women appears to be independent of the extent of left ventricular systolic dysfunction or remodelling compared with men.


The influence of CD25+ cells on the generation of immunity to tumour cell lines in mice.
CD25+ regulatory T cells comprise 5-10% of CD4+ T cells in naïve mice and have been shown in several in vivo murine models to prevent the induction of autoimmune disease and inflammatory disease. Since T cells, which mediate autoimmunity, can through recognition of self-antigens also target tumour cells, it was postulated that CD25+ regulatory cells would also inhibit the generation of immune responses to rumours. Depletion of these cells using CD25-specific monoclonal antibodies has indeed been shown to promote rejection of several transplantable murine tumour cell lines including melanoma, leukaemia and colorectal carcinoma. Results obtained using these models indicate that in the absence of regulatory cells, CD4+ T cells mediate tumour immunity, although the precise mechanisms through which these cells result in tumour rejection have not yet been elucidated. The target antigens recognized by these CD4+ T cells have also not yet been identified. Immunization of mice with tumour cells in the absence of CD25+ regulatory cells does, however, induce immunity against a variety of different tumour cell lines indicating that the target antigen(s) are shared amongst tumours of distinct histological origins. Since CD25+ regulatory cells have been identified in humans, the possibility that the cells inhibit immune responses to shared rejection antigens expressed by human tumours is worthy of investigation.


Behavioral evaluation of male and female mice pups exposed to fluoxetine during pregnancy and lactation.
Fluoxetine (FLX) has been widely prescribed for depression during pregnancy and/or lactation. Since serotonin is a neurotrophic factor, the use of FLX by mothers could disrupt brain development resulting in behavioral alterations in their progeny. This study evaluated the effects of developmental FLX exposure on anxiety, depression, aggressivity and pain sensitivity of male and female mice pups. Swiss dams were treated daily, by gavage, with 7.5 mg/kg of FLX during pregnancy and lactation. Pups were submitted to open-field, forced swimming, elevated plus-maze, intruder-resident and hot plate tests at adolescence and adulthood. In male pups, exposure to FLX decreased ambulation at postnatal day (PND) 40 and tended (p=0.07) to increase the latency to the first attack in the intruder-resident test at PND 70, suggesting decreased impulsivity. In female pups, FLX exposure increased immobility time in the forced swimming test at both PND 30 and 70, which is interpreted as depressive-like behavior. In conclusion, our results suggest that maternal exposure to FLX during pregnancy and lactation results in enduring behavioral alterations in male and female pups throughout life.


Influence of magnesium ions on heat shock and ethanol stress responses of Saccharomyces cerevisiae.
This study has highlighted the role of magnesium ions in the amelioration of the detrimental effects of ethanol toxicity and temperature shock in a winemaking strain of Saccharomyces cerevisiae. Specifically, results based on measurements of cellular viability and heat shock protein synthesis together with scanning electron microscopy have shown that, by increasing the bioavailability of magnesium ions, physiological protection is conferred on yeast cells. Elevating magnesium levels in the growth medium from 2 to 20 mM results in repression of certain heat shock proteins following a typical heat shock regime (30-42 degrees C shift). Seed inocula cultures prepropagated in elevated levels of magnesium (i.e. 'preconditioned') also conferred thermotolerance on cells and repressed the biosynthesis of heat shock proteins. Similar results were observed in response to ethanol stress. Extra- and intracellular magnesium may both act in the physiological stress protection of yeast cells and this approach offers potential benefits in alcoholic fermentation processes. The working hypothesis based on our findings is that magnesium protects yeast cells by preventing increases in cell membrane permeability elicited by ethanol and temperature-induced stress.


Potassium containing toothpastes for dentine hypersensitivity.
Dentine hypersensitivity may be defined as the pain arising from exposed dentine, typically in response to external stimuli, and which cannot be explained by any other form of dental disease. Many treatment regimens have been recommended over the years, and in recent years particular attention has been focused on toothpastes containing various potassium salts. To compare the effectiveness of potassium containing toothpastes with control toothpastes in reducing dentine hypersensitivity. The following databases were searched: Cochrane Oral Health Group Trials Register (searched until August 2005); CENTRAL (until August 2005); EMBASE/MEDLINE, PubMed, Web of Science (until September 2005). Bibliographies of clinical studies and reviews identified in the electronic search were checked for studies published outside the electronically searched journals. Randomised controlled trials (RCTs) in which the effect on dentine hypersensitivity of potassium containing toothpastes was tested against non-potassium containing control toothpastes. Two of the review authors independently recorded the results of the included trials using a specially designed form. Sensitivity was assessed by using thermal, tactile, air blast, and subjective methods. Six studies were included in the meta-analysis which showed the statistically significant effect of potassium nitrate toothpaste on air blast and tactile sensitivity at the 6 to 8 weeks follow up, e.g. the meta-analysis of air blast sensitivity showed a standardized mean difference in sensitivity score of -1.25 (95% CI: -1.65 to -0.851) in favour of treatment. The subjective assessment failed to show a significant effect at the 6 to 8 week assessment. The evidence generated by this review is based on a small number of individuals. Furthermore, the effect varies with the methods applied for assessing the sensitivity. Thus no clear evidence is available for the support of potassium containing toothpastes for dentine hypersensitivity.


Socially induced serotonergic fluctuations in the male auditory midbrain correlate with female behavior during courtship.
Cues from social partners trigger the activation of socially responsive neuromodulatory systems, priming brain regions including sensory systems to process these cues appropriately. The fidelity with which neuromodulators reflect the qualities of ongoing social interactions in sensory regions is unclear. We addressed this issue by using voltammetry to monitor serotonergic fluctuations in an auditory midbrain nucleus, the inferior colliculus (IC), of male mice (Mus musculus) paired with females, and by concurrently measuring behaviors of both social partners. Serotonergic activity strongly increased in male mice as they courted females, relative to serotonergic activity in the same males during trials with no social partners. Across individual males, average changes in serotonergic activity were negatively correlated with behaviors exhibited by female partners, including broadband squeaks, which relate to rejection of males. In contrast, serotonergic activity did not correlate with male behaviors, including ultrasonic vocalizations. These findings suggest that during courtship, the level of serotonergic activity in the IC of males reflects the valence of the social interaction from the perspective of the male (i.e., whether the female rejects the male or not). As a result, our findings are consistent with the hypothesis that neuromodulatory effects on neural responses in the IC may reflect the reception, rather than the production, of vocal signals.


Cytokinesis blocked micronucleus assay of peripheral lymphocytes revealing the genotoxic effect of formaldehyde exposure.
Formaldehyde (FA), which is said to be a carcinogenic agent, is commonly used in anatomy laboratories. This study used the cytokinesis blocked micronucleus assay (CBMN) to assess DNA damage due to FA exposure by measuring the frequency of micronuclei (MN) in lymphocytes. The extent of DNA damage was assessed with respect to the duration of exposure. Thirty male anatomy laboratory workers from various medical colleges involved with storing specimens and embalming were included in the study. Thirty males who were not exposed to FA were included as a comparison group. Blood samples were collected after informed consent was given. Information regarding age, duration of FA exposure and smoking habits was obtained by a questionnaire. The CBMN assay was conducted on cultured isolated lymphocytes stained with Giemsa. MN were counted in a total of 1000 binucleated lymphocytes. The effect of smoking was assessed using appropriate statistical tests. The frequency of MN in lymphocytes was significantly higher in the exposed group (P < 0.001). The duration of exposure correlated positively with the frequency of MN (r = 0.5, P = 0.02). Neither aging nor smoking correlated significantly with the formation of MN. The present study highlights significant DNA damage in people exposed to FA. The extent of damage was directly proportional to the duration of exposure.


Is contracting a form of privatization?
Contracting is often seen as a form of privatization, with contracts functioning as the tool that makes privatization possible. But contracting is also viewed by some as a means for the private sector to expand in a covert way its presence within the health sector. This article discusses the wider meaning of the term privatization in the health sector and the ways in which it is achieved. Privatization is seen here not simply as an action that leads to a new situation but also as one that leads to a change in behaviour. It is proposed that privatization may be assessed by looking at the ownership, management, and mission or objectives of the entity being privatized. Discussed also is the use of contracting by the state as a tool for state interventionism that is not based on authoritarian regulation.


Genetic and molecular characterization of multiflorous spikelet in oat.
Multiflorous spikelets are found in several grass species of agricultural and economic interest. In oat, this morphological characteristic is associated with the production of naked grains. Although many genetic studies have been performed over the past century, the inheritance of the multiflorous spikelet trait is not fully understood in oat. The objectives of this study were to evaluate environmental effects on the multiflorous spikelet trait, to estimate the number of genes controlling the trait, and to clone and characterize sequences of the AP2 gene in oat. Two genetic populations of recombinant inbreed lines were screened for the multiflorous spikelet trait from different years and sowing dates under field experiments. Normal, multiflorous, and mosaic spikelets were analyzed in the whole panicle for both years and sowing dates. Specific primer pairs for the AP2 gene was utilized to amplify and clone oat sequences. The results demonstrate that under higher temperature and day-length conditions, the variable expressivity of the multiflorous spikelet trait was less pronounced in both populations. Genetic analyses indicated the action of one major gene and two or three modifying genes controlling the expression of the multiflorous spikelet trait in oat, depending on the genetic background. Sequences with similarity to the AP2 gene were isolated from the oat lines UFRGS 017004-2 and URS Taura, and genetic polymorphisms were identified, which are valuable to confirm the action of AP2 on the multiflorous spikelet trait. Our results provide information to assist in the development of future studies of the multiflorous spikelet trait in oat.


Natural and managed watersheds show similar responses to recent climate change.
Changes in climate are driving an intensification of the hydrologic cycle and leading to alterations of natural streamflow regimes. Human disturbances such as dams, land-cover change, and water diversions are thought to obscure climate signals in hydrologic systems. As a result, most studies of changing hydroclimatic conditions are limited to areas with natural streamflow. Here, we compare trends in observed streamflow from natural and human-modified watersheds in the United States and Canada for the 1981-2015 water years to evaluate whether comparable responses to climate change are present in both systems. We find that patterns and magnitudes of trends in median daily streamflow, daily streamflow variability, and daily extremes in human-modified watersheds are similar to those from nearby natural watersheds. Streamflow in both systems show negative trends throughout the southern and western United States and positive trends throughout the northeastern United States, the northern Great Plains, and southern prairies of Canada. The trends in both natural and human-modified watersheds are linked to local trends in precipitation and reference evapotranspiration, demonstrating that water management and land-cover change have not substantially altered the effects of climate change on human-modified watersheds compared with nearby natural watersheds.


Natural selection on immune responsiveness in blue tits Parus caeruleus.
What is the form of natural selection on immune responsiveness? For a population at evolutionary equilibrium, there are two different scenarios. First, it is generally assumed that immune defense has both benefits and costs. If variation in immune responsiveness is due to variation in how individuals trade off these costs and benefits, one would expect immune responsiveness to be subject to stabilizing selection. Second, it is well known that an individual's immune responsiveness is often dependent on its overall condition. If immune responsiveness is condition-dependent, one would expect immune responsiveness to be under positive directional selection. We would therefore expect that the form of natural selection on immune responsiveness depends on the relative magnitude of these two sources of variation: variation in how individuals trade off the costs and benefits of defense, and variation in condition. We measured primary and secondary antibody responsiveness to diphtheria-tetanus vaccine in blue tits during winter and investigated the relationship between responsiveness and survival to the following breeding season. We use responsiveness to these antigens as measures of an individual's ability or propensity to mount an antibody response in case of an infection. Interestingly, different measures of responsiveness were subject to different selective regimes: primary responsiveness to diphtheria was subject to stabilizing selection, whereas secondary responsiveness to tetanus was subject to positive directional selection. In contrast, there was no significant selection on primary responsiveness to tetanus or secondary responsiveness to diphtheria. The finding of stabilizing selection on a measure of responsiveness is evidence that immune defense can incur fitness costs; a central but little-tested assumption of theories of the ecology and evolution of immunological defense. The finding of directional selection on a measure of responsiveness is consistent with the idea that immune responsiveness is condition-dependent, although we cannot rule out the alternative explanation that the population is not at evolutionary equilibrium with respect to this trait.


Msx1 disruption leads to diencephalon defects and hydrocephalus.
We have analyzed the expression of the Msx1 gene in the developing mouse brain and examined the brain phenotype in homozygotes. Msx1 is expressed in every cerebral vesicle throughout development, particularly in neuroepithelia, such as those of the fimbria and the medulla. Timing analysis suggests that Msx1(nLacZ) cells delaminate and migrate radially from these epithelia, mainly at embryonic days 14-16, while immunohistochemistry studies reveal that some of the beta-galactosidase migrating cells are oligodendrocytes or astrocytes. Our results suggest that the Msx1 neuroepithelia of fimbria and medulla may be a source of glial precursors. The Msx1 mutants display severe hydrocephalus at birth, while the subcommissural organ, the habenula, and the posterior commissure fail to develop correctly. No label was detected in the mutant subcommissural organ using a specific antibody against Reissner's fiber. Besides, the fasciculus retroflexus deviates close to the subcommissural organ, while the paraventricular thalamic nucleus shows histological disorganization. Our results implicate the Msx1 gene in the differentiation of the subcommissural organ cells and posterior commissure and that Msx1 protein may play a role in the pathfinding and bundling of the fasciculus retroflexus and in the structural arrangement of the paraventricular thalamic nucleus.


Activation and binding of C3 by Candida albicans.
Interaction with components of the complement system is an important aspect of the pathogenesis of infection by Candida albicans. The key role of C3 as an opsonic ligand and as an element in amplification of complement activation led us to examine several factors that influence the activation and binding of C3 cleavage fragments to the yeast. Activation and binding of C3 were determined by use of normal human serum containing 125I-labeled C3. Incubation of yeast-phase cells in 20% serum led to deposition of 2.5 X 10(5) to 3.0 X 10(5) molecules of C3 per yeast cell. Binding of C3 was absent in serum that was heat inactivated for 30 min at 37 degrees C or in serum that was chelated with EDTA. Chelation of serum with EGTA [ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid] reduced binding of C3 fragments by 31%. These results suggest that the alternative complement pathway is the primary mechanism for activation and binding of C3 fragments to C. albicans. Bound C3 could be partially removed (50%) by treatment with 1.0 M hydroxylamine. In contrast, 1.0 M hydroxylamine removed 98% of the C3 fragments bound to encapsulated Cryptococcus neoformans. These results suggest that ester bonds are the primary mechanism for binding C3 to C. neoformans, whereas binding of C3 to C. albicans involves both ester and amide bonds. Monoclonal antibodies specific for C3c and an iC3b neoantigen were used to identify the fragment of C3 that was bound to C. albicans. The results showed that the primary fragment bound to the yeast was C3b. An examination of the kinetics of activation and binding of C3 fragments showed that activation and binding were very rapid. Near-maximal binding occurred after a 2.5- to 5-min incubation period. In contrast, activation and binding of C3 fragments to C. neoformans proceeded at a slower rate, with maximal binding requiring 10 to 20 min. These results indicate that activation and binding of C3 fragments by the yeasts C. albicans and C. neoformans differ in several important characteristics.


Extracellular vesicle profiling and their use as potential disease specific biomarker.
Cell-derived vesicles in particular extracellular vesicles (EVs) such as microparticles (MPs) and microvesicles besides exosomes are raising more and more attention as a novel and unique approach to detect diseases. It has recently become apparent that disease specific MP signatures or profiles might be beneficial to differentiate chronic liver diseases such as non-alcoholic fatty liver disease and chronic hepatitis C, to monitor their progression or possibly to assess treatment outcome. Therefore EVs might serve as a novel inexpensive and minimally invasive method to screen risk patients for the outbreak of a disease even before the initial symptoms, to follow up treatment complications and disease relapse. The purpose of the current review is to summarize already published EVs signatures for a limited number of exemplary diseases and to discuss their possible impact. Additionally, it will be discussed if the combination of EV profiling and miRNA profiling could be a future joint tool for the purpose of detecting cancer and from far larger interest to ultimately distinguish among various tumor entities. EVs might increase the chance of early detection of chronic diseases or cancers especially if applied as part of yearly health screenings in the future.


Finding Nemo's Genes: A chromosome-scale reference assembly of the genome of the orange clownfish Amphiprion percula.
The iconic orange clownfish, Amphiprion percula, is a model organism for studying the ecology and evolution of reef fishes, including patterns of population connectivity, sex change, social organization, habitat selection and adaptation to climate change. Notably, the orange clownfish is the only reef fish for which a complete larval dispersal kernel has been established and was the first fish species for which it was demonstrated that antipredator responses of reef fishes could be impaired by ocean acidification. Despite its importance, molecular resources for this species remain scarce and until now it lacked a reference genome assembly. Here, we present a de novo chromosome-scale assembly of the genome of the orange clownfish Amphiprion percula. We utilized single-molecule real-time sequencing technology from Pacific Biosciences to produce an initial polished assembly comprised of 1,414 contigs, with a contig N50 length of 1.86 Mb. Using Hi-C-based chromatin contact maps, 98% of the genome assembly were placed into 24 chromosomes, resulting in a final assembly of 908.8 Mb in length with contig and scaffold N50s of 3.12 and 38.4 Mb, respectively. This makes it one of the most contiguous and complete fish genome assemblies currently available. The genome was annotated with 26,597 protein-coding genes and contains 96% of the core set of conserved actinopterygian orthologs. The availability of this reference genome assembly as a community resource will further strengthen the role of the orange clownfish as a model species for research on the ecology and evolution of reef fishes.


Badges to Acknowledge Open Practices: A Simple, Low-Cost, Effective Method for Increasing Transparency.
Beginning January 2014, Psychological Science gave authors the opportunity to signal open data and materials if they qualified for badges that accompanied published articles. Before badges, less than 3% of Psychological Science articles reported open data. After badges, 23% reported open data, with an accelerating trend; 39% reported open data in the first half of 2015, an increase of more than an order of magnitude from baseline. There was no change over time in the low rates of data sharing among comparison journals. Moreover, reporting openness does not guarantee openness. When badges were earned, reportedly available data were more likely to be actually available, correct, usable, and complete than when badges were not earned. Open materials also increased to a weaker degree, and there was more variability among comparison journals. Badges are simple, effective signals to promote open practices and improve preservation of data and materials by using independent repositories.


Questionnaire surveys: methodological and epistemological problems for field-based ethnopharmacologists.
The classical scientific approach is empirical. One of the favoured means of gathering quantitative data in the health and social sciences, including ethnopharmacology and medical ethnobotany, is by use of questionnaires. However, while there are numerous published articles discussing the importance of questionnaire content, the fact that questionnaires themselves may be inappropriate in a number of cultural contexts, even where literacy is not a factor, is usually ignored. In this paper, the authors will address the main issues posed by the use of questionnaire surveys, using case studies based on their own personal experiences as ethnopharmacologists 'in the field'. The pros and cons of qualitative and quantitative research and the use of alternative means to elicit quantitative data will be discussed.


Endothelial modulation of pH-dependent pressor response in isolated perfused rabbit lungs.
With the use of isolated perfused rabbit lungs (n = 152), roles of endothelium-derived relaxing factor (EDRF) in pulmonary vascular responses to hypocapnia and hypercapnia were studied. Lungs were ventilated with a gas mixture containing 1, 5, or 10% CO2 and 21% O2, adjusting the perfusate pH to 7.8, 7.4, or 7.1, respectively. Methemoglobin (MetHb), hemoglobin (Hb), methylene blue (MB), and L-argininosuccinic acid (L-ASA) were used as modulators of EDRF. To eliminate augmented shear stress, we used papaverine during hypercapnia. As a measure of EDRF, we spectrophotometrically examined nitric oxide (NO) metabolites in the perfusate. Hypocapnia and hypercapnia evoked, respectively, unsustainable vasodilatation and vasoconstriction. Hb, MB, and L-ASA, but not MetHb, produced an increase in baseline pulmonary arterial pressure (Ppa). These agents also exacerbated vasoconstriction during hypercapnia. Hypercapnia and hypocapnia caused an increase and decrease, respectively, in EDRF production. L-ASA suppressed EDRF production in hypercapnic lungs. Papaverine did not suppress EDRF production under hypercapnia. In conclusion, 1) the effects of pH on pulmonary circulation are transient, 2) the increase in Ppa caused by hypercapnia is modulated by EDRF, and 3) the pulmonary EDRF genesis is activated by hypercapnic acidosis but suppressed by hypocapnic alkalosis.


Relationship between individual differences in functional connectivity and facial-emotion recognition abilities in adults with traumatic brain injury.
Although several studies have demonstrated that facial-affect recognition impairment is common following moderate-severe traumatic brain injury (TBI), and that there are diffuse alterations in large-scale functional brain networks in TBI populations, little is known about the relationship between the two. Here, in a sample of 26 participants with TBI and 20 healthy comparison participants (HC) we measured facial-affect recognition abilities and resting-state functional connectivity (rs-FC) using fMRI. We then used network-based statistics to examine (A) the presence of rs-FC differences between individuals with TBI and HC within the facial-affect processing network, and (B) the association between inter-individual differences in emotion recognition skills and rs-FC within the facial-affect processing network. We found that participants with TBI showed significantly lower rs-FC in a component comprising homotopic and within-hemisphere, anterior-posterior connections within the facial-affect processing network. In addition, within the TBI group, participants with higher emotion-labeling skills showed stronger rs-FC within a network comprised of intra- and inter-hemispheric bilateral connections. Findings indicate that the ability to successfully recognize facial-affect after TBI is related to rs-FC within components of facial-affective networks, and provide new evidence that further our understanding of the mechanisms underlying emotion recognition impairment in TBI.


Mammary ductoscopy for evaluation of nipple discharge.
Most breast cancers originate in the ductal epithelium with normal cells progressing to atypia and finally to carcinoma. Ductoscopy enables one to visualize and sample the ductal epithelium and therefore identify early changes cytologically. This report describes our experience with mammary ductoscopy as a tool for evaluation of nipple discharge at Beth Israel Medical Center. A prospective review of all patients who have undergone ductoscopy for evaluation of persistent nipple discharge was performed. The Acueity ductoscopy system with .9-mm scope and a video monitor with x60 magnification were used. Brush biopsy samples and lavage fluid were obtained from some patients and were sent for cytologic analysis. A subset of patients underwent ductoscopically guided duct excision. Ninety-three patients underwent ductoscopic evaluation of 110 ducts. Of these, 67 patients had abnormal findings and therefore underwent ductoscopically guided duct excision. The remaining 26 patients (28%) had normal ductoscopic examinations. The depth at which intraductal abnormalities were visualized was from 3 to 8 cm with an average of 4.4 cm for cancer cases and from 1 to 10 cm with an average of 4.5 cm for papillomas. Forty-two patients were diagnosed with papilloma/papillomatosis, six patients were diagnosed with atypical papilloma/atypical ductal hyperplasia/atypical lobular hyperplasia, and six patients were diagnosed with cancer. Of the six patients diagnosed with cancer, 67% had normal breast imaging, and other than nipple discharge, 67% had normal breast examinations. Mammary ductoscopy is a useful tool in the evaluation of patients with nipple discharge. Although the most common cause of nipple discharge is an intraductal papilloma, nipple discharge can be the presenting symptom for cancer. Our experience revealed a papilloma rate of 45% (42 of 93), cancer rate of 6.5% (6 of 93), and an atypia rate of 6.5% (6 of 93) among the patients with nipple discharge. Mammary ductoscopy allows for accurate visualization, analysis, and excision of intraductal abnormalities. Many deeper intraductal abnormalities could be missed by blind surgical excision.


Formulation and evaluation of transdermal nanogel for delivery of artemether.
Artemether (ART) is second to artesunate in being the most widely used derivatives of artemisinin in combination therapy of malaria. Nanostructured lipid carrier (NLC) formulations were prepared following our previous report using optimized ART concentration of 0.25 g dissolved in 5% w/v mixture of solid (Gelucire 43/01 and Phospholipon 85G) and liquid (Transcutol) lipids at 90 °C. An aqueous surfactant phase at 90 °C was added (dropwise) under magnetic stirring (1000 rpm) for 5 min. The pre-emulsion was speedily homogenized at 28,000 rpm for 15 min and further probe sonicated at 60% amplitude (15 min). Resultant sample was cooled at room temperature and frozen at - 80 °C prior to lyophilization. The freeze-dried sample was used for solid-state characterization as well as in the formulation of transdermal nanogels using three polymers (Carbopol 971P, Poloxamer 407, and Prosopis africana peel powder) to embed the ART-NLC, using ethanol as a penetration enhancer. Transdermal ART-nanogels were characterized accordingly (physical examination, pH, drug content, rheology, spreadability, stability, particle size and morphology, skin irritation, in vitro and ex vivo skin permeation, and analysis of permeation data), P < 0.05. Results indicated that ART nanogels showed good encapsulation, drug release, pH-dependent swelling, stability, and tolerability. Overall, ART nanogels prepared from Poloxamer 407 showed the most desirable drug permeation, pH, swellability, spreadability, viscosity, and transdermal antiplasmodial properties superior to PAPP-ANG > C971P-ANG. A two-patch/week concurrent application of the studied nanogels could offer 100% cure of malaria as a lower-dose (50 mg ART) patient-friendly regimen devoid of the drug's many side effects.


Preoperative anemia: Predictor of free flap reconstruction complications in head and neck cancer.
Free flap reconstruction has been the mainstay among reconstruction surgeries for head and neck cancer. Intraoperative and postoperative hemoglobin (Hb) levels were both possible risk factors of flap failure and had been discussed widely. However, few investigations of preoperative Hb were seen in the previous study with its effect to flap condition remain uncertain and no conclusions in the literature. Patients who underwent free flap reconstruction after head and neck surgery in our institution between May 2014 and May 2019 were enrolled. The postoperative flap condition was observed carefully, and re-exploration was performed if necessary. We then retrospectively collected patient data with several intraoperative and postoperative indices. A total of 598 patients were enrolled in our study. The total major flap complication rate was 10.6%, with an overall success rate of 89.4%. They were predominantly male (95%), and most of them underwent free flap reconstruction for the first time (91%). A total of 81 (13%) patients received radiotherapy before reconstruction. Among all factors, the preoperative Hb level and free flap type showed significance in univariate and multivariate analyses. A previous history of radiotherapy, body mass index, nutrition status, or poorly controlled diabetes mellitus showed no significant results in either univariate or multivariate analysis. Our study showed that a lower preoperative Hb level affects free flap survival. Meanwhile, preoperative radiotherapy history has no significant influence in either univariate or multivariate analysis.


[Cyclic thrombocytopenia complicated with Sjögren syndrome and the mechanism of periodic platelet count fluctuation].
Cyclic thrombocytopenia is a rare disorder with periodic changes of the platelet count. We experienced a patient with cyclic thrombocytopenia and Sjögren syndrome (SS), and studied the mechanism of the cyclic changes of the platelet count. The patient, a 75-year-old woman, was referred to our hospital because of dry mouth, dry eyes, and severe thrombocytopenia. Her platelet count varied from 0.2 x 10(4)/microl to 22.7 x 10(4)/microl in 14-28-day cycles. Anti-SS-A/Ro antibody and Schirmer's test were positive. Histological examination of the salivary glands revealed infiltration of inflammatory cells, leading to the diagnosis of SS. There was an inverse relation between the platelet count and serum levels of PA-IgG. When the platelet count was low, the number of bone marrow megakaryocytes and the colony counts of CFU-Meg decreased. The serum thrombopoietin level increased at the nadir of the platelet count and decreased as platelets increased. After prednisolone therapy, her platelet count increased along with the improvement of sicca symptoms. These findings suggest that platelet fluctuation is due to the periodic increase of platelet destruction, as well as periodic failure of platelet production.


Comparing Neuromorphic Solutions in Action: Implementing a Bio-Inspired Solution to a Benchmark Classification Task on Three Parallel-Computing Platforms.
Neuromorphic computing employs models of neuronal circuits to solve computing problems. Neuromorphic hardware systems are now becoming more widely available and "neuromorphic algorithms" are being developed. As they are maturing toward deployment in general research environments, it becomes important to assess and compare them in the context of the applications they are meant to solve. This should encompass not just task performance, but also ease of implementation, speed of processing, scalability, and power efficiency. Here, we report our practical experience of implementing a bio-inspired, spiking network for multivariate classification on three different platforms: the hybrid digital/analog Spikey system, the digital spike-based SpiNNaker system, and GeNN, a meta-compiler for parallel GPU hardware. We assess performance using a standard hand-written digit classification task. We found that whilst a different implementation approach was required for each platform, classification performances remained in line. This suggests that all three implementations were able to exercise the model's ability to solve the task rather than exposing inherent platform limits, although differences emerged when capacity was approached. With respect to execution speed and power consumption, we found that for each platform a large fraction of the computing time was spent outside of the neuromorphic device, on the host machine. Time was spent in a range of combinations of preparing the model, encoding suitable input spiking data, shifting data, and decoding spike-encoded results. This is also where a large proportion of the total power was consumed, most markedly for the SpiNNaker and Spikey systems. We conclude that the simulation efficiency advantage of the assessed specialized hardware systems is easily lost in excessive host-device communication, or non-neuronal parts of the computation. These results emphasize the need to optimize the host-device communication architecture for scalability, maximum throughput, and minimum latency. Moreover, our results indicate that special attention should be paid to minimize host-device communication when designing and implementing networks for efficient neuromorphic computing.


Prospective assessment of overactive bladder symptoms in women who have undergone transvaginal surgery for advanced vaginal wall prolapse: a preliminary report.
To evaluate the changes in symptoms of overactive bladder (OAB) on the advanced cystocele after restoration of prolapse using the Perigee system. This prospective clinical study recorded 16 women for the Perigee procedure due to advanced cystocele with OAB symptoms. Prolapse assessment was measured according to the pelvic organ prolapse quantization system. Complete urodynamic examination was performed before and six months after the operation. Subjects were also asked to fill in a questionnaire before and after the operation for a quantitative assay of OAB symptoms. Additional pelvic reconstructive surgeries including the Apogee procedure, retropubic tension-free vaginal tape and posterior colporrhaphy with tension-free vaginal tape were performed concurrently on 10 patients. Eleven cases were available for data analysis. Results showed significant improvement at points Aa and Ba after the operation. The Perigee procedure could restore the advanced cystocele adequately, with points Aa and Ba almost back to their normal physiological and anatomical positions. However, urodynamics showed two subjects with motor urgency and one with sensory urgency having objective improvement in urge sensation. Questionnaire results of postoperative subjective symptoms revealed a significant improvement with respect to coping, concern and sleep as compared to preoperative symptoms. This preliminary report reveals that the Perigee procedure is an efficient and effective surgical approach for the treatment of anterior vaginal wall prolapse. It can also improve the subjective symptoms of OAB.


Determining barriers and facilitators to engagement for families in a family-based, multicomponent healthy lifestyles intervention for children and adolescents: a qualitative study.
Recruitment and retention in child and adolescent healthy lifestyle intervention services for childhood obesity is challenging, and inequalities across social groups are persistent. This study aimed to understand the barriers and facilitators to engagement in a multicomponent assessment-and-intervention healthy lifestyle programme for children and their families, based in the home and community. Qualitative interview-based study of past users (n=76) of a family-based multicomponent healthy lifestyle programme in a mixed urban-rural region of New Zealand. Semistructured, home-based interviews were conducted and thematically analysed with peer debriefing for validity. Families were selected through stratified random sampling to include a range of levels of engagement, including those who declined their referral, with equal numbers of interviews with Indigenous and non-Indigenous families. Three interactive and compounding determinants were identified as influencing engagement in Whānau Pakari: acute and chronic life stressors, societal norms of weight and body size and historical experiences of healthcare. These determinants were present across societal, system and healthcare service levels. A negative referral experience to Whānau Pakari often resulted in participants declining further input or disengaging from the programme. A fourth domain, respectful and compassionate healthcare, was identified as a mitigator of these three themes, facilitating participant engagement despite previous negative experiences. While participant engagement in healthy lifestyle programmes is affected by determinants which appear to operate outside immediate service provision, the programme is an opportunity to acknowledge past instances of stigma and the wider challenges of healthy lifestyle change. The experience of the referral to Whānau Pakari is important for setting the scene for future engagement in the programme. Respectful, compassionate care is critical to enhanced retention in multidisciplinary healthy lifestyle programmes and ongoing engagement in healthcare services overall.


Taking a bite out of nutrition and arbovirus infection.
Nutrition is a key factor in host-pathogen defense. Malnutrition can increase both host susceptibility and severity of infection through a number of pathways, and infection itself can promote nutritional deterioration and further susceptibility. Nutritional status can also strongly influence response to vaccination or therapeutic pharmaceuticals. Arthropod-borne viruses (arboviruses) have a long history of infecting humans, resulting in regular pandemics as well as an increasing frequency of autochthonous transmission. Interestingly, aside from host-related factors, nutrition could also play a role in the competence of vectors required for transmission of these viruses. Nutritional status of the host and vector could even influence viral evolution itself. Therefore, it is vital to understand the role of nutrition in the arbovirus lifecycle. This Review will focus on nutritional factors that could influence susceptibility and severity of infection in the host, response to prophylactic and therapeutic strategies, vector competence, and viral evolution.


Validation of the Greek version of the Bergen Social Media Addiction Scale in Undergraduate Students.
Social Networking Sites (SNSs) number at least 3.81 billion users worldwide, while in Greece, 74.7% of internet users resort to SNSs. However, there is no instrument adapted to the Greek language that assesses addiction to SNSs. The Bergen Social Media Addiction Scale is a short 6-item questionnaire that assesses social media addiction. This study aimed to evaluate the psychometric properties of the Greek version of the Bergen Social Media Addiction Scale. The sample consisted of 325 University students (mean age 21.6 years, SD=5.26). Confirmatory factor analysis was performed that supported the one-factor model. The reliability of the tool was tested with various indicators and was found to be satisfactory. The regression analysis identified gender, time of SNSs use, number of SNSs held by an individual and stress as statistically significant predictors of SNSs addiction. Positive correlations were found between SNSs addiction and stress, depression, anxiety and loneliness, while negative correlations were observed between SNSs addiction, self-esteem and age. Stress fully mediated the relationship between depression and SNSs addiction. The findings of the present study indicate that the Greek version of the Bergen Social Media Addiction Scale is a valid and reliable instrument for assessing SNSs addiction. The relationship between SNSs addiction and the biopsychosocial model was confirmed.


Pathogenesis of post-thymectomy autoimmunity. Role of syngeneic MLR-reactive T cells.
Thymectomy of 3-day-old mice results in the development of multi-organ-specific autoimmune diseases. The disease process is mediated by CD4+ T cells and is characterized by an inflammatory infiltrate in the affected organ(s) and the presence of autoantibodies. Our analysis of the phenotype of the CD4+ T cells that remain in the 3-day thymectomized animal revealed that the majority (approximately 80%) of the CD4+ lymph node cells express an activated (MEL-14low) phenotype and a smaller percentage expressed the T cell activation Ag CD69 and IL-2R alpha-chain. Thymectomized animals also had an increase in the frequency of mitogen-induced CD4+ IL-4 producers and significantly higher levels of total serum IgG. Functional studies demonstrated that lymph node T cells from 3-day thymectomized mice had an enhanced response in the syngeneic MLR and appeared to preferentially respond to syngeneic dendritic cells. To determine whether the syngeneic MLR-reactive T cells were involved in the pathogenesis of the organ-specific disease, we developed a model that mimicked the 3dTx model by grafting neonatal thymi to adult nu/nu recipients followed by removal of the thymus graft on day 3 or 4. When compared with mice transplanted with an untreated thymus, nu/nu mice transplanted with adult APC-containing thymi demonstrated a decrease in the incidence and severity of gastritis, a marked decrease in the titer of anti-parietal cell Ab, and a decrease in total serum IgG. Thus, intrathymic tolerization to complexes of self-peptides and MHC class II on adult APC prevents organ-specific autoimmune disease.


Dynamics of genomic clones in breast cancer patient xenografts at single-cell resolution.
Human cancers, including breast cancers, comprise clones differing in mutation content. Clones evolve dynamically in space and time following principles of Darwinian evolution, underpinning important emergent features such as drug resistance and metastasis. Human breast cancer xenoengraftment is used as a means of capturing and studying tumour biology, and breast tumour xenografts are generally assumed to be reasonable models of the originating tumours. However, the consequences and reproducibility of engraftment and propagation on the genomic clonal architecture of tumours have not been systematically examined at single-cell resolution. Here we show, using deep-genome and single-cell sequencing methods, the clonal dynamics of initial engraftment and subsequent serial propagation of primary and metastatic human breast cancers in immunodeficient mice. In all 15 cases examined, clonal selection on engraftment was observed in both primary and metastatic breast tumours, varying in degree from extreme selective engraftment of minor (<5% of starting population) clones to moderate, polyclonal engraftment. Furthermore, ongoing clonal dynamics during serial passaging is a feature of tumours experiencing modest initial selection. Through single-cell sequencing, we show that major mutation clusters estimated from tumour population sequencing relate predictably to the most abundant clonal genotypes, even in clonally complex and rapidly evolving cases. Finally, we show that similar clonal expansion patterns can emerge in independent grafts of the same starting tumour population, indicating that genomic aberrations can be reproducible determinants of evolutionary trajectories. Our results show that measurement of genomically defined clonal population dynamics will be highly informative for functional studies using patient-derived breast cancer xenoengraftment.


General and abdominal obesity and risk of esophageal and gastric adenocarcinoma in the European Prospective Investigation into Cancer and Nutrition.
General obesity, as reflected by BMI, is an established risk factor for esophageal adenocarcinoma (EAC), a suspected risk factor for gastric cardia adenocarcinoma (GCC) and appears unrelated to gastric non-cardia adenocarcinoma (GNCC). How abdominal obesity, as commonly measured by waist circumference (WC), relates to these cancers remains largely unexplored. Using measured anthropometric data from 391,456 individuals from the European Prospective Investigation into Cancer and Nutrition (EPIC) study and 11 years of follow-up, we comprehensively assessed the association of anthropometric measures with risk of EAC, GCC and GNCC using multivariable proportional hazards regression. One hundred twenty-four incident EAC, 193 GCC and 224 GNCC were accrued. After mutual adjustment, BMI was unrelated to EAC, while WC showed a strong positive association (highest vs. lowest quintile HR = 1.19; 95% CI, 0.63-2.22 and HR = 3.76; 1.72-8.22, respectively). Hip circumference (HC) was inversely related to EAC after controlling for WC, while WC remained positively associated (HR = 0.35; 0.18-0.68, and HR=4.10; 1.94-8.63, respectively). BMI was not associated with GCC or GNCC. WC was related to higher risks of GCC after adjustment for BMI and more strongly after adjustment for HC (highest vs. lowest quintile HR = 1.91; 1.09-3.37, and HR = 2.23; 1.28-3.90, respectively). Our study demonstrates that abdominal, rather than general, obesity is an indisputable risk factor for EAC and also provides evidence for a protective effect of gluteofemoral (subcutaneous) adipose tissue in EAC. Our study further shows that general obesity is not a risk factor for GCC and GNCC, while the role of abdominal obesity in GCC needs further investigation.


Evidence for in vivo growth potential and vascular remodeling of tissue-engineered artery.
Nondegradable synthetic polymer vascular grafts currently used in cardiovascular surgery have no growth potential. Tissue-engineered vascular grafts (TEVGs) may solve this problem. In this study, we developed a TEVG using autologous bone marrow-derived cells (BMCs) and decellularized tissue matrices, and tested whether the TEVGs exhibit growth potential and vascular remodeling in vivo. Vascular smooth muscle-like cells and endothelial-like cells were differentiated from bone marrow mononuclear cells in vitro. TEVGs were fabricated by seeding these cells onto decellularized porcine abdominal aortas and implanted into the abdominal aortas of 4-month-old, bone marrow donor pigs (n = 4). Eighteen weeks after implantation, the dimensions of TEVGs were measured and compared with those of native abdominal aortas. Expression of molecules associated with vascular remodeling was examined with reverse transcription-polymerase chain reaction assay and immunohistochemistry. Eighteen weeks after implantation, all TEVGs were patent with no sign of thrombus formation, dilatation, or stenosis. Histological and immunohistochemical analyses of the retrieved TEVGs revealed regeneration of endothelium and smooth muscle and the presence of collagen and elastin. The outer diameter of three of the four TEVGs increased in proportion to increases in body weight and outer native aorta diameter. Considerable extents of expression of molecules associated with extracellular matrix (ECM) degradation (i.e., matrix metalloproteinase and tissue inhibitor of matrix metalloproteinase) and ECM precursors (i.e., procollagen I, procollagen III, and tropoelastin) occurred in the TEVGs, indicating vascular remodeling associated with degradation of exogenous ECMs (implanted decellularized matrices) and synthesis of autologous ECMs. This study demonstrates that the TEVGs with autologous BMCs and decellularized tissue matrices exhibit growth potential and vascular remodeling in vivo of tissue-engineered artery.


Antimicrobial de-escalation in the critically ill patient and assessment of clinical cure: the DIANA study.
The DIANA study aimed to evaluate how often antimicrobial de-escalation (ADE) of empirical treatment is performed in the intensive care unit (ICU) and to estimate the effect of ADE on clinical cure on day 7 following treatment initiation. Adult ICU patients receiving empirical antimicrobial therapy for bacterial infection were studied in a prospective observational study from October 2016 until May 2018. ADE was defined as (1) discontinuation of an antimicrobial in case of empirical combination therapy or (2) replacement of an antimicrobial with the intention to narrow the antimicrobial spectrum, within the first 3 days of therapy. Inverse probability (IP) weighting was used to account for time-varying confounding when estimating the effect of ADE on clinical cure. Overall, 1495 patients from 152 ICUs in 28 countries were studied. Combination therapy was prescribed in 50%, and carbapenems were prescribed in 26% of patients. Empirical therapy underwent ADE, no change and change other than ADE within the first 3 days in 16%, 63% and 22%, respectively. Unadjusted mortality at day 28 was 15.8% in the ADE cohort and 19.4% in patients with no change [p = 0.27; RR 0.83 (95% CI 0.60-1.14)]. The IP-weighted relative risk estimate for clinical cure comparing ADE with no-ADE patients (no change or change other than ADE) was 1.37 (95% CI 1.14-1.64). ADE was infrequently applied in critically ill-infected patients. The observational effect estimate on clinical cure suggested no deleterious impact of ADE compared to no-ADE. However, residual confounding is likely.