Myelin restoration: progress and prospects for human cell replacement therapies.
Oligodendrocytes are the primary source of myelin in the adult central nervous system (CNS), and their dysfunction or loss underlies several diseases of both children and adults. Dysmyelinating and demyelinating diseases are thus attractive targets for cell-based strategies since replacement of a single presumably homogeneous cell type has the potential to restore functional levels of myelin. To understand the obstacles that cell-replacement therapy might face, we review oligodendrocyte biology and emphasize aspects of oligodendrocyte development that will need to be recapitulated by exogenously transplanted cells, including migration from the site of transplantation, axon recognition, terminal differentiation, axon wrapping, and myelin production and maintenance. We summarize studies in which different types of myelin-forming cells have been transplanted into the CNS and highlight the continuing challenges regarding the use of cell-based therapies for human white matter disorders.


Relationship of A1 segment hypoplasia to anterior communicating artery aneurysm morphology and risk factors for aneurysm formation.
OBJECTIVE Hypoplasia of the A1 segment of the anterior cerebral artery is frequently observed in patients with anterior communicating artery (ACoA) aneurysms. The effect of this anatomical variant on ACoA aneurysm morphology is not well understood. METHODS Digital subtraction angiography images were reviewed for 204 patients presenting to the authors' institution with either a ruptured or an unruptured ACoA aneurysm. The ratio of the width of the larger A1 segment to the smaller A1 segment was calculated. Patients with an A1 ratio greater than 2 were categorized as having A1 segment hypoplasia. The relationship of A1 segment hypoplasia to both patient and aneurysm characteristics was then assessed. RESULTS Of 204 patients that presented with an ACoA aneurysm, 34 (16.7%) were found to have a hypoplastic A1. Patients with A1 segment hypoplasia were less likely to have a history of smoking (44.1% vs 62.9%, p = 0.0410). ACoA aneurysms occurring in the setting of a hypoplastic A1 were also found to have a larger maximum diameter (mean 7.7 vs 6.0 mm, p = 0.0084). When considered as a continuous variable, increasing A1 ratio was associated with decreasing aneurysm dome-to-neck ratio (p = 0.0289). There was no significant difference in the prevalence of A1 segment hypoplasia between ruptured and unruptured aneurysms (18.9% vs 10.7%; p = 0.1605). CONCLUSIONS Our results suggest that a hypoplastic A1 may affect the morphology of ACoA aneurysms. In addition, the relative lack of traditional risk factors for aneurysm formation in patients with A1 segment hypoplasia argues for the importance of hemodynamic factors in the formation of ACoA aneurysms in this anatomical setting.


2,4-dienoyl-CoA reductase regulates lipid homeostasis in treatment-resistant prostate cancer.
Despite the clinical success of Androgen Receptor (AR)-targeted therapies, reactivation of AR signalling remains the main driver of castration-resistant prostate cancer (CRPC) progression. In this study, we perform a comprehensive unbiased characterisation of LNCaP cells chronically exposed to multiple AR inhibitors (ARI). Combined proteomics and metabolomics analyses implicate an acquired metabolic phenotype common in ARI-resistant cells and associated with perturbed glucose and lipid metabolism. To exploit this phenotype, we delineate a subset of proteins consistently associated with ARI resistance and highlight mitochondrial 2,4-dienoyl-CoA reductase (DECR1), an auxiliary enzyme of beta-oxidation, as a clinically relevant biomarker for CRPC. Mechanistically, DECR1 participates in redox homeostasis by controlling the balance between saturated and unsaturated phospholipids. DECR1 knockout induces ER stress and sensitises CRPC cells to ferroptosis. In vivo, DECR1 deletion impairs lipid metabolism and reduces CRPC tumour growth, emphasizing the importance of DECR1 in the development of treatment resistance.


Post-traumatic stress disorder (PTSD) symptoms in children with severe epilepsy.
To assess symptoms of post-traumatic stress disorder (PTSD) in children with severe epilepsy and the associations of trauma symptoms across age, comorbid symptoms, epilepsy-specific factors, parental resources, and psychopathology. Fifty children with severe epilepsy across three different age groups (0-5 yrs., 6-12 yrs., 13-18 yrs.) were assessed with developmental-sensitive and standardized PTSD assessment tools when hospitalized at the tertiary epilepsy center Filadelfia, Denmark. The Diagnostic Infant and Preschool Assessment (DIPA), the Darryl test, and the ITQ questionnaire were used to assess the three age groups, respectively. Twenty-two percent of the overall sample met the criteria for PTSD, with a prevalence of symptoms increasing with age (6%, 28%, and 40%). Comorbid psychiatric symptoms in preschoolers were present in 81% of the children witnessing a high level of distress in this group. Behavioral difficulties were elevated across all three age groups, and 40% of the children with trauma symptoms had a parent with concurrent psychopathology. To the authors' knowledge, this study is the first to assess trauma symptoms with standardized tests in children with more complicated epilepsies. Trauma symptoms in the group are high; however, there is a need for larger scale studies and research into trauma symptoms in children with more severe epilepsy than those assessable with the included assessment tools. The trauma perspective in severe childhood epilepsy might further clarify the complex associations of biological and contextual variables that affect the children's life quality and enable better preventative treatment options for this group.


Regulation of apoptosis by a prostate-specific and prostate cancer-associated noncoding gene, PCGEM1.
PCGEM1 is a prostate tissue-specific, and prostate cancer-associated noncoding RNA (ncRNA) gene. Previous results revealed a significant association of elevated PCGEM1 expression levels in prostate cancer cells of African-American patients, whose mortality rate is the highest among prostate cancer patients. Functional study of PCGEM1 demonstrated a marked increase in colony formation in LNCaP prostate cancer cells and NIH3T3 mouse fibroblast cells. This study demonstrates that PCGEM1 overexpression in LNCaP cell culture model results in the inhibition of apoptosis induced by doxorubicin (DOX). Induction of p53 and p21(Waf1/Cip1) by DOX were delayed in LNCaP cells stably overexpressing PCGEM1 (LNCaP-PCGEM1 cells) compared to control LNCaP cells. The protein levels of cleaved caspase 7, and cleaved PARP were attenuated in DOXtreated LNCaP-PCGEM1 cells compared to control LNCaP cells. Similar results were observed in LNCaP cells transiently overexpressing PCGEM1. The inhibition of PARP cleavage by PCGEM1 overexpression was also observed in LNCaP-PCGEM1 cells incubated with etoposide and sodium selenite. Fluorescence-Activated Cell Sorter Annexin-V analysis revealed significantly lower percentage of apoptotic cells in DOX-treated LNCaP-PCGEM1 cells compared to control LNCaP cells. The attenuation of apoptic response appears to be androgen dependent in this experimental model, as androgen-independent variants of LNCaP cells did not exhibit this response. In summary, this study provides new insights into cell biologic functions and novel features of an ncRNA. Further, these data unravel biological mechanisms of cell growth/cell survival-associated functions of this ncRNA in a widely used prostate cancer cell culture model.


Pathological evaluation of tumor grade for salivary adenoid cystic carcinoma: A proposal of an objective grading system.
Three pathological grading systems advocated by Perzin/Szanto, Spiro, and van Weert are currently used for adenoid cystic carcinoma (AdCC). In these systems, the amount or presence of the solid tumor component in AdCC specimens is an important index. However, the "solid tumor component" has not been well defined. Salivary AdCC cases (N = 195) were collected after a central pathology review. We introduced a novel criterion for solid tumor component, minAmax (minor axis maximum). The largest solid tumor nest in each AdCC case was histologically screened, the maximum oval fitting the solid nest was estimated, and the length of the minor axis of the oval (minAmax) was measured. The prognostic cutoff for the minAmax was determined using training and validation cohorts. All cases were evaluated for the four grading systems, and their prognostic impact and interobserver variability were examined. The cutoff value for the minAmax was set at 0.20 mm. Multivariate prognostic analyses showed the minAmax and van Weert systems to be independent prognostic tools for overall, disease-free, and distant metastasis-free survival while the Perzin/Szanto and Spiro systems were selected for overall survival but not for disease-free or distant metastasis-free survival. The highest hazard ratio for overall survival (11.9) was obtained with the minAmax system. The reproducibility of the minAmax system (kappa coefficient of 0.81) was scored as very good while those of the other three systems were scored as moderate. In conclusion, the minAmax is a simple, objective, and highly reproducible grading system useful for prognostic stratification for salivary AdCC.


SU-E-T-529: Dosimetric Evaluation with Heterogeneity in Acuros XB Advanced Dose Calculation Algorithm and Anisotropic Analytical Algorithm (AAA).
To evaluate accuracy of Acuros XB Advanced Dose Calculation Algorithm and Anisotropic Analytical Algorithm (AAA) with measurements in predicting doses beyond air gaps. Three virtual phantoms with layers were created in Eclipse Treatment Planning System. Each layer in phantoms was assigned in terms of water (top), air (middle) and water (bottom) medium. Central axis depth dose in water (bottom medium) beyond 2, 4 and 6 cm air gaps were computed for 5 cm water equivalent material positioned before air gaps. Dose computation was performed for 6MV photon beam with 3×3 and 5×5 cm2 field sizes at 100 cm SSD to surface of phantoms using AAA_10.0.28 and Acuros XB_10.0.28. Next, solid water and Styrofoam were manufactured to mimic virtual phantoms. By keeping identical field, beam parameters, and geometries that were used for dose computation, 100 MUs were delivered to phantoms, and measurements at selected depths were acquired with cylindrical ionization chamber. Measured central axis depth doses were compared against calculated central axis depth doses computed from Acuros XB and AAA. Acuros XB predicted doses within ±2% of measured doses except at 1cm depth in phantoms with 4 cm air gap (-2.4%) and 6 cm air gap (-2.8%) for field size 3×3cm2 . Acuros XB showed better dose prediction compared to AAA at all measured depths. Smallest test field size in phantom with largest air gap produced highest range (between depths 1 and 5 cm) in percentage dose difference (AAA: -2.9% to -9.9% and Acuros XB: -2.8% to 1.5%). Improper modeling of primary beam attenuation or lateral scatter (or combination of both) within air gap may have resulted into dose discrepancies. The findings suggest Acuros XB is more accurate to use in dose predictions when tumor is located beyond small air cavity (heterogeneity) within the patient.


Trophic action of leptin on hypothalamic neurons that regulate feeding.
In adult mammals, the adipocyte-derived hormone leptin acts on the brain to reduce food intake by regulating the activity of neurons in the arcuate nucleus of the hypothalamus (ARH). Here, we report that neural projection pathways from the ARH are permanently disrupted in leptin-deficient (Lepob/Lepob) mice and leptin treatment in adulthood does not reverse these neuroanatomical defects. However, treatment of Lepob/Lepob neonates with exogenous leptin rescues the development of ARH projections, and leptin promotes neurite outgrowth from ARH neurons in vitro. These results suggest that leptin plays a neurotrophic role during the development of the hypothalamus and that this activity is restricted to a neonatal critical period that precedes leptin's acute regulation of food intake in adults.


Studies of twins: what can they tell us about the fetal origins of adult disease?
There has been much interest in evidence that people with lower birthweight have higher risk of adult cardiovascular disease, but the causal pathways underlying such observations are uncertain. Study of twins offers an opportunity to shed light on the underlying causal pathways, in particular by investigating the role of 'shared' factors vs. factors affecting each individual fetus. This involves comparing results of within-cohort vs. within-pair analyses. Twins share many factors during gestation but birthweight discordance (difference in birthweight within a twin pair) cannot be determined by these shared factors and must relate to factors affecting growth of each individual fetus. If associations seen in a cohort of twins remain in within-pair analyses, then factors specific to each individual must be involved in the underlying causal pathways. Conversely, if the relationships disappear or substantially diminish in within-pair analyses, then factors common to the pair must be involved. Comparison of findings in monozygotic vs. dizygotic twins may provide insights into the role of genetic factors, although issues related to chorionicity need to be taken into account. We tabulate published data and conclude that differences in methodology and analyses preclude informative meta-analysis, and that analysis of pooled data would provide more useful information.


Effects of study area size on geographic characterizations of health events: prostate cancer incidence in Southern New England, USA, 1994-1998.
We consider how representations of geographic variation in prostate cancer incidence across Southern New England, USA may be affected by selection of study area and/or properties of the statistical analysis. A spatial scan statistic was used to monitor geographic variation among 35,167 incident prostate cancer cases diagnosed in Massachusetts, Connecticut and Rhode Island from 1994 to 1998, in relation to the 1990 populations of men 20+ years of age living in that region. Results from the combined-states analysis were compared to those from single-states. Impact of scanning procedures set to examine up to 50% or no more than 10% of at-risk populations also was evaluated. With scanning set to 50%, 5 locations in the combined-states analysis were identified with markedly distinct incidence rates. Fewer than expected cases were estimated for nearly all Connecticut, Rhode Island and West Central Massachusetts, whereas census tracts on and around Cape Cod, and areas of Southwestern Connecticut and adjacent to greater Boston were estimated to have yielded more than expected incidence. Results of single-state analyses exhibited several discrepancies from the combined-states analysis. More conservative scanning found many more locations with varying incidence, but discrepancies between the combined- and single-state analysis were fewer. It is important to acknowledge the conditional nature of spatial analyses and carefully consider whether a true cluster of events is identified or artifact stemming from selection of study area size and/or scanning properties.


Physical activity and breast cancer risk.
Data from a multicentric case-control study on breast cancer conducted in Italy were used to analyze the relationship of occupational and leisure-time physical activity with breast cancer risk. Cases were 2569 histologically confirmed incident breast cancer cases, and controls were 2588 patients admitted to the same network of hospitals of cases for acute, nonneoplastic, nonhormone related diseases. After allowance for major identified potential confounding factors (including an estimate of total calorie intake), the odds ratios (ORs) were 0.70, 0.71, 0.64, and 0.54 in subsequent levels of physical activity at work at ages 30-39, compared to the lowest level. The association was similar for occupational physical activity at ages 15-19 and still apparent at ages 50-59, with risk estimates of 0.86, 0.85, 0.85, and 0.62. The ORs for the highest versus the lowest category of leisure-time physical activity were also below unity (ORs for the highest level of leisure-time physical activity at ages 15-19, 0.95; at ages 30-39, 0.76; and at ages 50-59, 0.66). The protection of physical activity was apparently stronger below age 60 at diagnosis and was consistent across the strata of selected covariates, although the protection was somewhat greater for more educated women.


The impact of COVID-19 non-pharmaceutical interventions on the lived experiences of people living in Thailand, Malaysia, Italy and the United Kingdom: A cross-country qualitative study.
This qualitative study explores the impact of non-pharmaceutical interventions (NPIs), including social distancing, travel restrictions and quarantine, on lived experiences during the first wave of the COVID-19 pandemic in Thailand (TH), Malaysia (MY), Italy (IT) and the United Kingdom (UK). A total of 86 interviews (TH: n = 28; MY: n = 18; IT: n = 20; UK: n = 20) were conducted with members of the public, including healthcare workers (n = 13). Participants across countries held strong views on government imposed NPIs, with many feeling measures lacked clarity. Most participants reported primarily negative impacts of NPIs on their lives, including through separation, isolation and grief over missed milestones; work-related challenges and income loss; and poor mental health and wellbeing. Nonetheless, many also experienced inadvertent positive consequences, including more time at home to focus on what they most valued in life; a greater sense of connectedness; and benefits to working life. Commonly employed coping strategies focused on financial coping (e.g. reducing spending); psycho-emotional coping (e.g. engaging in spiritual practices); social coping and connectedness (e.g., maintaining relationships remotely); reducing and mitigating risks (e.g., changing food shopping routines); and limiting exposure to the news (e.g., checking news only occasionally). Importantly, the extent to which participants' lived experiences were positive or negative, and their ability to cope was underpinned by individual, social and economic factors, with the analysis indicating some salient differences across countries and participants. In order to mitigate negative and unequal impacts of NPIs, COVID-19 policies will benefit from paying closer attention to the social, cultural and psychological-not just biological-vulnerabilities to, and consequences of public health measures.


Climate warming causes life-history evolution in a model for Atlantic cod (Gadus morhua).
Climate change influences the marine environment, with ocean warming being the foremost driving factor governing changes in the physiology and ecology of fish. At the individual level, increasing temperature influences bioenergetics and numerous physiological and life-history processes, which have consequences for the population level and beyond. We provide a state-dependent energy allocation model that predicts temperature-induced adaptations for life histories and behaviour for the North-East Arctic stock (NEA) of Atlantic cod (Gadus morhua) in response to climate warming. The key constraint is temperature-dependent respiratory physiology, and the model includes a number of trade-offs that reflect key physiological and ecological processes. Dynamic programming is used to find an evolutionarily optimal strategy of foraging and energy allocation that maximizes expected lifetime reproductive output given constraints from physiology and ecology. The optimal strategy is then simulated in a population, where survival, foraging behaviour, growth, maturation and reproduction emerge. Using current forcing, the model reproduces patterns of growth, size-at-age, maturation, gonad production and natural mortality for NEA cod. The predicted climate responses are positive for this stock; under a 2°C warming, the model predicted increased growth rates and a larger asymptotic size. Maturation age was unaffected, but gonad weight was predicted to more than double. Predictions for a wider range of temperatures, from 2 to 7°C, show that temperature responses were gradual; fish were predicted to grow faster and increase reproductive investment at higher temperatures. An emergent pattern of higher risk acceptance and increased foraging behaviour was also predicted. Our results provide important insight into the effects of climate warming on NEA cod by revealing the underlying mechanisms and drivers of change. We show how temperature-induced adaptations of behaviour and several life-history traits are not only mediated by physiology but also by trade-offs with survival, which has consequences for conservation physiology.


Distraction osteogenesis after membranous bone onlay grafting in a dog model.
The purpose of this experiment was to study the possibility of distraction osteogenesis in a membranous bone onlay graft to the mandible and to clarify the histology of the bone repair. Four dogs, 5 months of age at the beginning of the experiment, were used for this study. The zygomatic arch was exposed in the subperiosteal plane, and a 3-cm long, full-thickness portion of the arch was harvested. The lateral surface of the mandibular body was exposed in the subperiosteal plane, and the bone was fixed to the lateral surface as a membranous onlay graft using screws. A vertical osteotomy through the graft and underlying mandibular body was done postoperatively at week 1 in dog 1, week 2 in dog 2, week 3 in dog 3, and week 4 in dog 4. An external distraction device was applied to the mandibular body, and distraction was started 7 days after the operation at a rate of 1 mm/d for 10 days. After completion of distraction, the device was left in place for 6 weeks to allow for bony consolidation. Radiographs were carried out at 2, 4, and 6 weeks postdistraction. All dogs were killed 6 weeks after distraction. New bone between the native underlying mandibular segments was generated in the distraction zone in all dogs. New bone was not generated between the segments of the membranous bone onlay graft in dog 1, but was generated in dog 2, dog 3, and dog 4. However, in dogs 2 and 3, the new bone between the segments was less firm, with more fibrous tissue, than the bone between the native underlying mandibular segments. Histologically, the distraction gap between the segments of the membranous bone onlay graft in dogs 2 and 3 was composed of considerable fibrous tissue in the central zone and activated osteoblastic cells forming new bone in the margins. In dog 4, there was much more osteoblastic activity in the distraction gap, and the new bone had the appearance of almost normal cortical bone. These findings show that distraction osteogenesis is possible in a membranous bone onlay graft and suggest that the distraction should be performed at least 4 weeks after the onlay grafting.


Self-Reported Food Hypersensitivity: Prevalence, Characteristics, and Comorbidities in the Norwegian Women and Cancer Study.
This study aims to investigate the prevalence of self-reported food hypersensitivity, (SFH), the characteristics of women with SFH, and whether SFH is associated with multiple health complaints among the participants of the Norwegian Women and Cancer study (NOWAC). We conducted a cross-sectional study among 64,316 women aged 41-76 years. The women were randomly selected from the Norwegian Central Person Register. Information on SFH and all covariates except age and place of residence was collected by questionnaires in 2002-2005. The prevalence of SFH in our study sample was 6.8% (95% confidence interval: 6.7-7.0). Logistic regression analysis showed a negative association between SFH and age (odds ratio [OR] 0.97). The odds of SFH increased among women living in or near urban centers, women with more than 9 years of education, women who did not have full-time work, women who had experienced poor economic conditions in childhood, those living without a partner, and those who did not consume alcohol or smoke (OR varied from 1.10 to 1.70). Women with a low body mass index had higher odds of SFH (OR 1.37) than those with a moderate body mass index. SFH was positively associated with poor self-perceived health (OR 2.56). The odds of SFH increased with the number of concurrent health complaints, with an OR for 5-6 comorbidities of 4.93. We found an association between SFH, poor health, and different socio demographic and lifestyle characteristics. Women with SFH had increased odds of reporting multiple health complaints.


Whole-body biodistribution and radiation dosimetry of 18F-GE067: a radioligand for in vivo brain amyloid imaging.
We have characterized the biodistribution and dosimetry of (18)F-3'-F-6-OH-BTA1 ((18)F-GE067), a newly developed radioligand to visualize and quantify amyloid burden, in healthy elderly human subjects. Six subjects (5 men and 1 woman; age range, 51-74 y) underwent dynamic whole-body PET/CT for 6 h after a bolus injection of (18)F-GE067. Source organs were delineated on PET/CT. Individual organ doses and effective doses were determined. No adverse events or clinically significant changes were observed. (18)F-GE067 is excreted predominantly through the hepatobiliary system. The gallbladder, upper large intestine, and small intestine are the organs with the highest absorbed dose (average, 287, 173, and 155 microGy/MBq, respectively). The mean effective dose was 33.8 +/- 3.4 microSv/MBq, a dose comparable to that of many other (18)F-labeled radiopharmaceuticals. The estimated effective dose of (18)F-GE067 for PET amyloid imaging was acceptable (class II-b defined by the World Health Organization), and relatively low variability between subjects was observed.


Evaluating an mHealth App for Health and Well-Being at Work: Mixed-Method Qualitative Study.
To improve workers' health and well-being, workplace interventions have been developed, but utilization and reach are unsatisfactory, and effects are small. In recent years, new approaches such as mobile health (mHealth) apps are being developed, but the evidence base is poor. Research is needed to examine its potential and to assess when, where, and for whom mHealth is efficacious in the occupational setting. To develop interventions for workers that actually will be adopted, insight into user satisfaction and technology acceptance is necessary. For this purpose, various qualitative evaluation methods are available. The objectives of this study were to gain insight into (1) the opinions and experiences of employees and experts on drivers and barriers using an mHealth app in the working context and (2) the added value of three different qualitative methods that are available to evaluate mHealth apps in a working context: interviews with employees, focus groups with employees, and a focus group with experts. Employees of a high-tech company and experts were asked to use an mHealth app for at least 3 weeks before participating in a qualitative evaluation. Twenty-two employees participated in interviews, 15 employees participated in three focus groups, and 6 experts participated in one focus group. Two researchers independently coded, categorized, and analyzed all quotes yielded from these evaluation methods with a codebook using constructs from user satisfaction and technology acceptance theories. Interviewing employees yielded 785 quotes, focus groups with employees yielded 266 quotes, and the focus group with experts yielded 132 quotes. Overall, participants muted enthusiasm about the app. Combined results from the three evaluation methods showed drivers and barriers for technology, user characteristics, context, privacy, and autonomy. A comparison between the three qualitative methods showed that issues revealed by experts only slightly overlapped with those expressed by employees. In addition, it was seen that the type of evaluation yielded different results. Findings from this study provide the following recommendations for organizations that are planning to provide mHealth apps to their workers and for developers of mHealth apps: (1) system performance influences adoption and adherence, (2) relevancy and benefits of the mHealth app should be clear to the user and should address users' characteristics, (3) app should take into account the work context, and (4) employees should be alerted to their right to privacy and use of personal data. Furthermore, a qualitative evaluation of mHealth apps in a work setting might benefit from combining more than one method. Factors to consider when selecting a qualitative research method are the design, development stage, and implementation of the app; the working context in which it is being used; employees' mental models; practicability; resources; and skills required of experts and users.


Neuroprotection by the selective iNOS inhibitor GW274150 in a model of Parkinson disease.
Neuroinflammation and the activation of inducible nitric oxide synthase (iNOS) have been proposed to play a role in the pathogenesis of Parkinson disease (PD). In this study we investigated the effects of the selective iNOS inhibitor GW274150 in the 6-OHDA model of PD. 6-OHDA administration was associated with increased numbers of cells expressing iNOS. Administration of the iNOS inhibitor twice daily for 7 days, beginning 2 days after the 6-OHDA lesioning, led to a significant neuroprotection as shown by assessment of the integrity of the nigrostriatal system by tyrosine hydroxylase immunocytochemistry and HPLC assessment of striatal dopamine content. However, GW274150 displayed a bell-shaped neuroprotective profile, being ineffective at high doses. 6-OHDA lesioning was associated with an increase in microglial activation as assessed by the MHC II antigen OX-6 and the number of matrix metalloproteinase 9 (MMP-9)-immunopositive cells. NO is a known modulator of MMP-9, and iNOS inhibition was associated with decreased numbers of MMP-9-immunopositive cells, culminating in a reduction in the numbers of reactive microglia. Withdrawal of GW274150 for a further 7 days negated any neuroprotective effects of iNOS inhibition, suggesting that the damaging effects of inflammation last beyond 7 days in this model and the continued administration of the drug may be required.


Intra-amygdala and systemic antagonism of NMDA receptors prevents the reconsolidation of drug-associated memory and impairs subsequently both novel and previously acquired drug-seeking behaviors.
The amygdala has long been considered a primary locus in mediating the effects of previously drug-associated stimuli on subsequent drug-seeking behavior, and the NMDA subtype of glutamate receptor within the amygdala is important for the consolidation of associations between environmental conditioned stimuli and the effects of addictive drugs. Here we demonstrate that amygdala NMDA receptors are also necessary for the reconsolidation of drug-associated memories. Using a behavioral task that specifically measures the conditioned reinforcing properties of a previously drug-paired stimulus, we show that infusion of the NMDA receptor antagonist D(-)-2-amino-5-phosphonopentanoic acid (D-APV) into the basolateral amygdala before a memory reactivation session disrupted the drug-associated memory and abolished subsequent instrumental responding for conditioned reinforcement. This effect was memory reactivation dependent, and the memory deficit persisted for at least 4 weeks. In contrast, infusion of d-APV immediately after the memory reactivation session had no effect on subsequent responding for conditioned reinforcement, indicating that NMDA receptors have a temporally limited role in the reconsolidation process. Furthermore, in molecular studies, we show that the reconsolidation-impairing effect of D-APV is correlated with downstream reductions in expression of the plasticity-related immediate early gene, zif268. We also demonstrate that systemic antagonism of NMDA receptors with MK-801 [(+)-5-methyl-10,11-dihydro-SH-dibenzo[a,d]cyclohepten-5,10-imine maleate] before memory reactivation subsequently reduced previously acquired instrumental drug-seeking behavior that depends on drug-associated cues acting as conditioned reinforcers. These data suggest that drugs modulating glutamatergic transmission at the NMDA receptor may be useful in the future treatment of relapse prevention in drug addiction through memory reconsolidation blockade.


High and low affinity IL 2 receptors: analysis by IL 2 dissociation rate and reactivity with monoclonal anti-receptor antibody PC61.
In this report we characterize two classes of interleukin 2 (IL 2) binding sites on the basis of their differential IL 2 dissociation rate and of their reactivity with PC61, a monoclonal anti-IL 2 receptor (IL 2-R) antibody. PC61 inhibited the binding of IL 2 to both classes of receptor, but IL 2 did not inhibit the binding of PC61. This indicates that PC61 recognizes a determinant that is distal to the actual IL 2 binding site of the receptor. Dissociation experiments showed that the addition of excess unlabeled IL 2 resulted in a biphasic release of radiolabeled IL 2; 80 to 90% was dissociated rapidly (dissociation half-time, t1/2 of 60 sec) and the remainder more slowly (t1/2 60 to 90 min). The proportion of high and low affinity IL 2-R, as well as the relative difference in dissociation rates fit very well with the estimates derived previously from Scatchard plot analysis of equilibrium IL 2 binding. The addition of PC61 caused an accelerated dissociation of IL 2 from both high and low affinity IL 2-R (t1/2 of 16 and 120 sec respectively).


Macdonald's model and the transmission of bilharzia.
The paper considers a model for the transmission of bilharzia based on Macdonald's assumptions, in the light of data observed in the field. It is shown, in particular, that the threshold parameter governing whether or not an endemic cycle can be established is closely related to the proportion of infected snails in a community, and that this proportion is normally observed to be rather smaller than is compatible with the model. By considering more sophisticated models, allowing for the latent period of infection in the snails, and also for spatial and seasonal heterogeneity, the effective proportion of infected snails, from the point of view of Macdonald's model, is shown to be rather larger, and expressions are given whereby it can be evaluated from observable quantities. However, for the data from Malirong which are taken as illustration, it is also demonstrated that an even more plausible threshold value is obtained from a simple model incorporating human immunity in addition to the assumptions of Macdonald's model, and that, if this model were reasonable, human immunity would appear to be the most important factor in controlling the level of the disease in Malirong.


The molecular basis of the effect of temperature on enzyme activity.
Experimental data show that the effect of temperature on enzymes cannot be adequately explained in terms of a two-state model based on increases in activity and denaturation. The Equilibrium Model provides a quantitative explanation of enzyme thermal behaviour under reaction conditions by introducing an inactive (but not denatured) intermediate in rapid equilibrium with the active form. The temperature midpoint (Teq) of the rapid equilibration between the two forms is related to the growth temperature of the organism, and the enthalpy of the equilibrium (DeltaHeq) to its ability to function over various temperature ranges. In the present study, we show that the difference between the active and inactive forms is at the enzyme active site. The results reveal an apparently universal mechanism, independent of enzyme reaction or structure, based at or near the active site, by which enzymes lose activity as temperature rises, as opposed to denaturation which is global. Results show that activity losses below Teq may lead to significant errors in the determination of DeltaG*cat made on the basis of the two-state ('Classical') model, and the measured kcat will then not be a true indication of an enzyme's catalytic power. Overall, the results provide a molecular rationale for observations that the active site tends to be more flexible than the enzyme as a whole, and that activity losses precede denaturation, and provide a general explanation in molecular terms for the effect of temperature on enzyme activity.


Risk of cervical intraepithelial neoplasia 2+ among women with a history of previous treatment for cervical intraepithelial neoplasia: ASCUS and LSIL Pap smears after treatment.
The objective of the current study was to describe outcomes among women with low-grade abnormalities on cervical cytology screening in the setting of previous excisional or ablative treatment for cervical intraepithelial neoplasia (CIN). Study participants were recruited into the Study to Understand Cervical Cancer Early Endpoints and Determinants. At enrollment, the patient's previous cytology results, previous colposcopic biopsy results, and previous cervical procedures were recorded. Study procedures included collection of biospecimens followed by colposcopy and biopsy. From clinical records, additional information regarding previous treatment for CIN was collected. Two hundred seventy-four women had an atypical squamous cells of uncertain significance (ASCUS) referral Pap and 532 women had a low-grade squamous intraepithelial lesion (LSIL) referral Pap. For patients with an ASCUS referral Pap, previous treatment was associated with an odds ratio for CIN 2+ (45.0% vs 28.2% of untreated patients) of 2.08 (95% confidence interval = 1.05-4.13, p = .04). For patients with an LSIL referral Pap, 33.3% of those women with previous treatment had CIN 2+ compared with 16.7% of those patients enrolled with no previous treatment (odds ratio = 2.49, 95% confidence interval = 1.12-5.51, p = .03). Patients with a history of previous treatment for CIN have a 2-fold risk of CIN 2+ at the time of colposcopy referral for ASCUS or LSIL cervical cytology.


GNE myopathy in Roma patients homozygous for the p.I618T founder mutation.
GNE myopathy is an autosomal-recessive disorder caused by mutations in the GNE gene, encoding the key enzyme in the sialic acid biosynthetic pathway, UDP-N-acetylglucosamine 2-epimerase/N-acetyl mannosamine kinase. We studied 50 Bulgarian Roma patients homozygous for p.I618T, an ancient founder mutation in the kinase domain of the GNE gene, dating before the Gypsy exodus from North West India. The clinical features in the Bulgarian GNE group can be described with disease onset mostly in the third decade, but in individual cases, onset was as early as 10 years of age. The majority of patients had foot drop as the first symptom, but three patients developed hand weakness first. Muscle weakness was early and severe for the tibialis anterior, and minimal or late for quadriceps femoris, and respiratory muscles were only subclinically affected even in the advanced stages of the disease. During a 15-year follow-up period, 32 patients became non-ambulant. The average period between disease onset and loss of ambulation was 10.34 ± 4.31 years, ranging from 3 to 20 years. Our analysis of affected sib pairs suggested a possible role of genetic modifying factors, accounting for significant variation in disease severity.


Tip-link protein protocadherin 15 interacts with transmembrane channel-like proteins TMC1 and TMC2.
The tip link protein protocadherin 15 (PCDH15) is a central component of the mechanotransduction complex in auditory and vestibular hair cells. PCDH15 is hypothesized to relay external forces to the mechanically gated channel located near its cytoplasmic C terminus. How PCDH15 is coupled to the transduction machinery is not clear. Using a membrane-based two-hybrid screen to identify proteins that bind to PCDH15, we detected an interaction between zebrafish Pcdh15a and an N-terminal fragment of transmembrane channel-like 2a (Tmc2a). Tmc2a is an ortholog of mammalian TMC2, which along with TMC1 has been implicated in mechanotransduction in mammalian hair cells. Using the above-mentioned two-hybrid assay, we found that zebrafish Tmc1 and Tmc2a can interact with the CD1 or CD3 cytoplasmic domain isoforms of Pcdh15a, and this interaction depends on the common region shared between the two Pcdh15 isoforms. Moreover, an interaction between mouse PCDH15-CD3 and TMC1 or TMC2 was observed in both yeast two-hybrid assays and coimmunoprecipitation experiments. To determine whether the Pcdh15-Tmc interaction is relevant to mechanotransduction in vivo, we overexpressed N-terminal fragments of Tmc2a in zebrafish hair cells. Overexpression of the Tmc2a N terminus results in mislocalization of Pcdh15a within hair bundles, together with a significant decrease in mechanosensitive responses, suggesting that a Pcdh15a-Tmc complex is critical for mechanotransduction. Together, these results identify an evolutionarily conserved association between the fish and mouse orthologs of PCDH15 and TMC1 and TMC2, supporting the notion that TMCs are key components of the transduction complex in hair cells.


Receptor-mediated endocytosis and intracellular trafficking of insulin and low-density lipoprotein by retinal vascular endothelial cells.
The authors investigated the receptor-mediated endocytosis (RME) and intracellular trafficking of insulin and low-density lipoprotein (LDL) in cultured retinal vascular endothelial cells (RVECs). Low-density lipoprotein and insulin were conjugated to 10 nm colloidal gold, and these ligands were added to cultured bovine RVECs for 20 minutes at 4 degrees C. The cultures were then warmed to 37 degrees C and fixed after incubation times between 30 seconds and 1 hour. Control cells were incubated with unconjugated gold colloid at times and concentrations similar to those of the ligands. Additional control cells were exposed to several concentrations of anti-insulin receptor antibody or a saturating solution of unconjugated insulin before incubation with gold insulin. Using transmission electron microscopy, insulin gold and LDL gold were both observed at various stages of RME. Insulin-gold particles were first seen to bind to the apical plasma membrane (PM) before clustering in clathrin-coated pits and internalization in coated vesicles. Gold was later visualized in uncoated cytoplasmic vesicles, corresponding to early endosomes and multivesicular bodies (MVBs) or late endosomes. In several instances, localized regions of the limiting membrane of the MVBs appeared coated, a feature of endosomal membranes not previously described. After RME at the apical PM and passage through the endosomal system, the greater part of both insulin- and LDL-gold conjugates was seen to accumulate in large lysosome-like compartments. However, a small but significant proportion of the internalized ligands was transcytosed and released as discrete membrane-associated quanta at the basal cell surface. The uptake of LDL gold was greatly increased in highly vacuolated, late-passage RVECs. In controls, anti-insulin receptor antibody and excess unconjugated insulin caused up to 89% inhibition in gold-insulin binding and internalization. These results illustrate the internalization and intracellular trafficking by RVECs of insulin and LDL through highly efficient RME, and they provide evidence for at least two possible fates for the endocytosed ligands. This study outlines a route by which vital macromolecules may cross the inner blood-retinal barrier.


Polyphenol, antioxidant and antimicrobial potential of six different white and red wine grape processing leftovers.
During winemaking, grape polyphenols are only partly extracted, and consequently unexploited. The main aim was to characterize the phenolic content of freeze-dried grape skin and seed (FDSS) extracts obtained from Slovenian and international grape varieties and to evaluate their antioxidant, antimicrobial and anti-adhesive activities. FDSS of six Vitis vinifera L. grapevine cultivars from Vipava Valley region (Slovenia) underwent extraction and sonification under different conditions. Flavonols were the predominant content of extracts from white 'Zelen' and 'Sauvignon Blanc' grape varieties, with strong antimicrobial activities against Gram-negative bacteria. 'Pinot Noir' FDSS extracted with 50% aqueous ethanol extraction produced a high phenolic content in the final extract, which was further associated with strong antioxidant and antimicrobial activities against all tested bacteria. Bacterial adhesion to stainless steel surfaces with minimal and maximal surface roughness was significantly inhibited (up to 60%) across a wide FDSS concentration range, with lower concentrations also effective with two types of stainless steel surfaces. FDSS extracts from winery by-products show interesting phenolic profiles that include flavonols, catechins, anthocyanins and hydroxycinnamic acids, with yields influenced by grapevine cultivar and extraction conditions. The antioxidant, antimicrobial and anti-adhesive activities of 50% aqueous ethanol 'Pinot Noir' FDSS extract reveals potential applications in food, pharmaceutical and cosmetic industries for these bioactive residues. © 2016 Society of Chemical Industry.


Epidermal growth factor receptor activation is localized within low-buoyant density, non-caveolar membrane domains.
Increasing evidence for the organization of cell-surface proteins and lipids into different detergent-insoluble rafts led us to investigate epidermal growth factor (EGF) receptor activation in the plasma membranes of A431 carcinoma cells, using a combination of cell fractionation and immunoprecipitation techniques. Density-gradient centrifugation of sodium carbonate cell extracts revealed that the vast majority of both stimulated and unstimulated EGF receptors were concentrated in a caveolin-rich light membrane (CLM) fraction, with the biochemical characteristics of detergent-insoluble glycolipid-rich domains (DIGs). However, ultrastructural analysis of the CLM fraction revealed that it contained a heterogeneous collection of vesicles, some with sizes greater than that expected for individual caveolae. Experiments with detergent-solubilized cells and isolated CLMs indicated that, in contrast with caveolin, EGF receptors were unlikely to be localized to DIG domains. Furthermore, immunoisolation of caveolin from CLMs revealed that EGF receptor activation occurs in a compartment distinct from caveolae. Similarly, using an anti-(EGF receptor) antibody, the bulk of the cellular caveolin was not co-immunoprecipitated from CLMs, thereby confirming that these two proteins reside in separate membrane domains. The deduction that caveolar signalling and EGF receptor activation occur in separable rafts argues for a multiplicity of signal transduction compartments within the plasma membrane. In addition, by demonstrating that EGF receptor activation is compartmentalized within low-density, non-caveolar regions of the plasma membrane, it is also shown that the co-localization of proteins in a CLM fraction is insufficient to prove caveolar localization.


Methane adsorption in ADOR zeolites: a combined experimental and DFT/CC study.
Physical adsorption of methane in purely siliceous molecular sieves prepared by a recently discovered synthetic pathway using 2D zeolites as nanoscale building blocks has been investigated by means of combined experimental and theoretical approaches. The DFT/CC-based method has been tested on ADOR zeolites of the UTL family and a few experimentally well-characterized siliceous zeolites. Excellent agreement between theoretical and experimental heats of adsorption has been found for OKO, PCR, MFI, CHA and AEI zeolites. The observed discrepancy for the UTL germanosilicate (2 kJ mol-1) has been plausibly explained using a simple model of D4R defects. The proposed methodology can be used as a reliable characterization tool for newly synthesized silica nanomaterials.


Requirement for the Drosophila COE transcription factor Collier in formation of an embryonic muscle: transcriptional response to notch signalling.
During Drosophila embryogenesis, mesodermal cells are recruited to form a stereotyped pattern of about 30 different larval muscles per hemisegment. The formation of this pattern is initiated by the specification of a special class of myoblasts, called founder cells, that are uniquely able to fuse with neighbouring myoblasts. We report here the role of the COE transcription factor Collier in the formation of a single muscle, muscle DA3([A])(DA4([T])). Col expression is first observed in two promuscular clusters (in segments A1-A7), the two corresponding progenitors and their progeny founder cells, but its transcription is maintained in only one of these four founder cells, the founder of muscle DA3([A]). This lineage-specific restriction depends on the asymmetric segregation of Numb during the progenitor cell division and involves the repression of col transcription by Notch signalling. In col mutant embryos, the DA3([A]) founder cells form but do not maintain col transcription and are unable to fuse with neighbouring myoblasts, leading to a loss-of-muscle DA3([A]) phenotype. In wild-type embryos, each of the DA3([A])-recruited myoblasts turns on col transcription, indicating that the conversion, by the DA3([A]) founder cell, of 'naive' myoblasts to express its distinctive pattern of gene expression involves activation of col itself. We find that muscles DA3([A]) and DO5([A]) (DA4([T]) and DO5([T])) derive from a common progenitor cell. Ectopic expression of Col is not sufficient, however, to switch the DO5([A]) to a DA3([A]) fate. Together these results lead us to propose that specification of the DA3([A]) muscle lineage requires both Col and at least one other transcription factor, supporting the hypothesis of a combinatorial code of muscle-specific gene regulation controlling the formation and diversification of individual somatic muscles.


An auditory repetition deficit under low memory load.
Previous studies of the auditory analogue of repetition blindness have led to different conclusions regarding the nature of the effect (e.g., N. Kanwisher & M. C. Potter, 1989; M. Miller & D. MacKay, 1994). In the present study, recall accuracy for repeated elements was examined with lists of 2 or 3 items presented dichotically under high temporal pressure. When this procedure was used, a repetition deficit in recall was obtained for both vowels (Experiment 1) and consonant-vowel syllables (Experiment 2). Further experiments demonstrated that this deficit decreases as the stimulus onset asynchrony between the 2 critical elements increases (Experiment 3) and showed that the effect also occurs for words and not just nonsense syllables (Experiment 4). In all 4 experiments, estimations of guessing biases showed that responses to unrepeated lists were not artificially favored over responses to repeated lists.


[Can finasteride reverse the progress of benign prostatic hyperplasia? A two-year placebo-controlled study.].
To study if placebo-induced improvement in men with symptomatic benign prostatic hyperplasia (BPH) is maintained over two years, and to study the efficacy and safety from intervention with finasteride 5 mg for 24 months. This was a multicenter, double-blind, placeba-controlled study involving 707 patients with moderate symptoms of BPH enrolled at 59 centers in five Scandinavian countries. Following enrollment and a four-week single-blind placebo run-in period, patients were randomized to receive finasteride 5 mg once daily or placebo for 24 months. Urinary symptoms, urinary flow rate, prostate volume, postvoiding residual urinary volume, and serum concentrations of prostate-specific antigen together with laboratory safety parameters were measured at entry and at months 12 and 24. Interim physical and laboratory examinations were performed when indicated clinically. In finasteride-treated patients the total symptom score improved throughout the study, with a significant difference between the two groups at 24 months (p<0.01) whereas in placebo-treated patients, there was an initial improvement in the symptom score but no change from baseline at 24 months. The maximum urinary flow rate decreased in the placebo group, but improved in the finasteride group, resulting in a between-group difference of 1.8 mL/s at 24 months (p<0.01). The mean change in prostate volume was +12% in the placebo group versus -19% in the finasteride-treated group (p%lt;0.01). Finasteride was generally well tolerated throughout the two-year study period. The efficacy of therapy with finasteride 5 mg in improving both symptoms and maximum urinary flow rate and reducing prostate volume has been shown to be maintained during 24 months while patients receiving placebo experienced a return to baseline or deterioration of these parameters during the study. These results demonstrate that finasteride can reverse the natural progression of BPH.


Mesenchymal Stromal Cells Are Retained in the Porcine Renal Cortex Independently of Their Metabolic State After Renal Intra-Arterial Infusion.
The regenerative capacities of mesenchymal stromal cells (MSCs) make them suitable for renal regenerative therapy. The most common delivery route of MSC is through intravenous infusion, which is associated with off-target distribution. Renal intra-arterial delivery offers a targeted therapy, but limited knowledge is available regarding the fate of MSCs delivered through this route. Therefore, we studied the efficiency and tissue distribution of MSCs after renal intra-arterial delivery to a porcine renal ischemia-reperfusion model. MSCs were isolated from adipose tissue of healthy male pigs, fluorescently labeled and infused into the renal artery of female pigs. Flow cytometry allowed MSC detection and quantification in tissue and blood. In addition, quantitative polymerase chain reaction was used to trace MSCs by their Y-chromosome. During infusion, a minor number of MSCs left the kidney through the renal vein, and no MSCs were identified in arterial blood. Ischemic and healthy renal tissues were analyzed 30 min and 8 h after infusion, and 1-4 × 104 MSCs per gram of tissue were detected, predominantly, in the renal cortex, with a viability >70%. Confocal microscopy demonstrated mainly glomerular localization of MSCs, but they were also observed in the capillary network around tubuli. The infusion of heat-inactivated (HI) MSCs, which are metabolically inactive, through the renal artery showed that HI-MSCs were distributed in the kidney in a similar manner to regular MSCs, suggesting a passive retention mechanism. Long-term MSC survival was analyzed by Y-chromosome tracing, and demonstrated that a low percentage of the infused MSCs were present in the kidney 14 days after administration, while HI-MSCs were completely undetectable. In conclusion, renal intra-arterial MSC infusion limited off-target engraftment, leading to efficient MSC delivery to the kidney, most of them being cleared within 14 days. MSC retention was independent of the metabolic state of MSC, indicating a passive mechanism.


The Gulf War Era Cohort and Biorepository: A Longitudinal Research Resource of Veterans of the 1990-1991 Gulf War Era.
The US Department of Veterans Affairs (VA) Gulf War Era Cohort and Biorepository (GWECB) is a nationally representative longitudinal cohort of US veterans who served during the 1990-1991 Gulf War era. The GWECB combines survey data, such as demographic, health behavior, and environmental exposure data; medical records; and a linked biorepository of blood specimens that can support a broad range of future research regarding health concerns unique to veterans of this era. To build this resource, the VA Cooperative Studies Program initiated a pilot study (2014-2016) to establish the GWECB and evaluate the processes required to build and maintain the resource. Participants (n = 1,275) consented to future sharing of their data and biospecimens for research purposes. Here we describe the pilot study, including recruitment and enrollment procedures, data collection and management, quality control, and challenges experienced. The GWECB data available to investigators under approved sharing mechanisms and the procedures for accessing them are extensively detailed. The study's consenting documents and a website link for the research survey are provided. Our hope is that new research drawing on the GWECB data and biospecimens will result in effective treatments and improved approaches to address the health concerns of Gulf War-era veterans.


Canadian survey of thyroid cancer.
We report the results of a multicentre retrospective chart review of 2214 patients with thyroid cancer registered at 13 radiotherapy centres between 1958 and 1978. The data analysed included sex, age at the time of diagnosis, pathological diagnosis, extent of disease before treatment, types of treatment and their complications, and the rates of recurrence and survival up to 24 years after diagnosis. Although papillary cancers were most common, anaplastic and miscellaneous tumours were more frequent than expected, which reflects the type of patients referred by endocrinologists and surgeons to radiotherapy centres. There were marked differences in patterns of referral to the centres. Some patients with papillary and follicular thyroid cancers died of these cancers up to 20 years after diagnosis. The clinical manifestations, treatment and outcome of the rarer types of thyroid malignant tumours were of particular interest. The influence of age at the time of diagnosis on survival rates for patients with papillary or follicular thyroid cancer was highly significant, indicating much more aggressive behaviour of these cancers in older patients, particularly those beyond the age of 60 years. A more detailed analysis of tumour subtypes should provide new information on their natural history and lead to better management.


Interpreting analyses of continuous covariates in affected sibling pair linkage studies.
Datasets collected for linkage analyses of complex human diseases often include a number of clinical or environmental covariates. In this study, we evaluated the performance of three linkage analysis methods when the relationship between continuous covariates and disease risk or linkage heterogeneity was modeled in three different ways: (1) The covariate distribution is determined by a quantitative trait locus (QTL), which contributes indirectly to the disease risk; (2) the covariate is not genetically determined, but influences the disease risk through statistical interaction with a disease susceptibility locus; (3) the covariate distribution differs in families linked or unlinked to a particular disease susceptibility locus. We analyzed simulated datasets with a regression-based QTL analysis, a nonparametric analysis of the binary affection status, and the ordered subset analysis (OSA). We found that a significant OSA result may be due to a gene that influences variability in the population distribution of a continuous disease risk factor. Conversely, a regression-based QTL analysis may detect the presence of gene-environment (GxE) interaction in a sample of primarily affected individuals. The contribution of unaffected siblings and the size of baseline lod scores may help distinguish between QTL and GxE models. As illustrated by a linkage study of multiplex families with age-related macular degeneration, our findings assist in the interpretation of analysis results in real datasets. They suggest that the side-by-side evaluation of OSA and QTL results may provide important information about the relationship of measured covariates with either disease risk or linkage heterogeneity.


Beneficial use of serum ferritin and heme oxygenase-1 as biomarkers in adult-onset Still's disease: A multicenter retrospective study.
Heme oxygenase (HO)-1 is a heme-degrading enzyme highly expressed in monocyte/macrophage, serum levels of which may be promising biomarker for adult-onset Still's disease (AOSD). We here report data on the use of serum ferritin and HO-1 levels in AOSD. Under the Hypercytokinemia Study Group collaboration, we collected sera from a total of 145 AOSD patients. Three independent experts judged whether the patients were definite AOSD depending on the clinical information. These 91 'definite AOSD' patients were further divided into active, remission, and relapse groups. Forty-six cases of systemic vasculitis, sepsis, etc. were included as disease controls. Serum ferritin and HO-1 levels were measured using ELISA. Associations between clinical symptoms, serum ferritin, and HO-1 were explored. Multivariate regression analysis was performed to identify independent variables associated with definite AOSD diagnosis. Serum ferritin and HO-1 levels were significantly higher in active and relapsed AOSD cases compared to disease controls, and were reduced by the treatment. Although a significant correlation was found between serum ferritin and HO-1 levels, a discrepancy was found in some cases such as iron-deficiency anemia. Receiver operating characteristic analysis identified optimal levels of serum ferritin (>819 ng/ml; sensitivity 76.1% and specificity 73.8%), and serum HO-1 (>30.2 ng/ml; sensitivity 84.8% and specificity 83.3%) that differentiated AOSD from controls. Interestingly, 88.9% of patients with AOSD who relapsed exceeded the cut-off value of serum HO-1 > 30.2 ng/ml, but only 50.0% exceeded serum ferritin >819 ng/ml (p = .013), suggesting that serum HO-1 levels may be a convenient indicator of AOSD disease status. Multivariate analysis identified neutrophilia, RF/ANA negativity, sore throat, and elevated serum HO-1 as independent variables associated with AOSD diagnosis. We confirmed that serum ferritin and HO-1 serve as highly specific and sensitive biomarkers for AOSD. A future prospective study with large sample size is necessary to determine whether these biomarkers could be included in Yamaguchi's Criteria.


Genetic polymorphisms associated with psoriasis and development of psoriatic arthritis in patients with psoriasis.
Psoriasis (PsO) is a chronic inflammatory disease with predominantly cutaneous manifestations. Approximately one third of patients with PsO develop psoriatic arthritis (PsA), whereas the remaining proportion of patients has isolated cutaneous psoriasis (PsC). These two phenotypes share common immunology, but with different heredity that might in part be explained by genetic variables. Using a candidate gene approach, we studied 53 single nucleotide polymorphisms (SNPs) in 37 genes that regulate inflammation. In total, we assessed 480 patients with PsO from DERMBIO, of whom 151 had PsC for 10 years or more (PsC10), 459 patients with PsA from DANBIO, and 795 healthy controls. Using logistic regression analysis, crude and adjusted for age and gender, we assessed associations between genetic variants and PsO, PsC10, and PsA, as well as associations between genetic variants and development of PsA in PsO. Eleven polymorphisms in 10 genes were nominally associated with PsO and/or PsC and/or PsA (P < 0.05). After correction for multiple testing with a false discovery rate of 5%, two SNPs remained significant: TNF (rs361525) was associated with PsO, PsC10, and PsA; and IL12B (rs6887695) was associated with PsO. Among a cohort of Danish patients with moderate-to-severe psoriasis, two SNPs in the IL12B and TNF genes were associated with susceptibility of psoriasis. None of the SNPs were specifically associated with isolated cutaneous psoriasis or psoriatic arthritis.


Factors associated with pulseless electric activity versus ventricular fibrillation: the Oregon sudden unexpected death study.
Corresponding with a continuing decline in the prevalence of sudden cardiac arrest cases presenting with ventricular fibrillation (VF), there has been a significant rise in the prevalence of pulseless electrical activity (PEA). Given significantly lower survival from PEA versus VF, we comprehensively investigated PEA correlates by incorporating first-responder data with lifetime clinical history information. In the Portland, Ore, metropolitan area (population ≈1 million), cases of out-of-hospital sudden cardiac arrest who underwent attempted resuscitation were identified prospectively (2002-2007). Those presenting with PEA versus VF and asystole were compared with χ² tests, ANOVA, and logistic regression. A total of 1277 cases aged ≥18 years underwent resuscitation by first responders (mean age, 65±16 years; 67% male). Presenting arrhythmia was VF in 48%, PEA in 25%, and asystole/other in the remainder. Compared with VF cases, PEA cases were older (mean age, 68 versus 63 years; P=0.0002), more likely to be female (37% versus 26%; P=0.0008), and less likely to survive to hospital discharge (6% versus 25%; P<0.0001). A history of syncope was strongly associated with PEA (odds ratio, 2.6; confidence interval, 1.3 to 5.3) after adjustment for age, gender, response time, and arrest circumstances. Black race was also independently associated with PEA (odds ratio, 2.6; confidence interval, 1.3 to 5.4). Pulmonary disease and female gender were significant factors associated with PEA (P for interaction=0.04). In a subgroup analysis of resting ECGs (n=391), there were no differences in cardiac clinical history or prevalence of cardiac conduction system disease (PEA, 31.6% versus VF, 32.2%; P=0.48). PEA cases had a significantly higher prevalence of syncope in their lifetime, with other correlates, including black race, that were distinct from VF cases. Potential mechanistic links between syncope and future manifestation with PEA warrant further exploration.


Efficient cell pairing in droplets using dual-color sorting.
The use of microfluidic droplets has become a powerful tool for the screening and manipulation of cells. However, currently this is restricted to assays involving a single cell type. Studies on the interaction of different cells (e.g. in immunology) as well as the screening of antibody-secreting cells in assays requiring an additional reporter cell, have not yet been successfully demonstrated. Based on Poisson statistics, the probability for the generation of droplets hosting exactly one cell of two different types is just 13.5%. To overcome this limitation, we have developed an approach in which different cell types are stained with different fluorescent dyes. Subsequent to encapsulation into droplets, the resulting emulsion is injected into a very compact sorting device allowing for analysis at high magnification and fixation of the cells close to the focal plane. By applying dual-color sorting, this furthermore enables the specific collection and analysis of droplets with exactly two different cells. Our approach shows an efficiency of up to 86.7% (more than 97% when also considering droplets hosting one or more cells of each type), and, hence, should pave the way for a variety of cell-based assays in droplets.


Cross-site comparison of gene expression data reveals high similarity.
Consistency and coherence of gene expression data across multiple sites depends on several factors such as platform (oligo, cDNA, etc.), environmental conditions at each laboratory, and data quality. The Hepatotoxicity Working Group of the International Life Sciences Institute Health and Environmental Sciences Institute consortium on the application of genomics to mechanism-based risk assessment is investigating these factors by comparing high-density gene expression data sets generated on two sets of RNA from methapyrilene (MP) experiments conducted at Abbott Laboratories and Boehringer-Ingelheim Pharmaceuticals, Inc. using a single platform (Affymetrix Rat Genome U34A GeneChip) at seven different sites. This article focuses on the evaluation of data quality and statistical models that facilitate the comparison of such data sets at the probe level. We present methods for exploring and quantitatively assessing differences in the data, with the principal goal being the generation of lists of site-insensitive genes responsive to low and high doses of MP. A combination of numerical and graphical techniques reveals important patterns and partitions of variability in the data, including the magnitude of the site effects. Although the site effects are significantly large in the analysis results, they appear to be primarily additive and therefore can be adjusted in the statistical calculations in a way that does not bias conclusions regarding treatment differences.


Versatile and Valuable Utilization of Amidohydrolase L-glutaminase in Pharma and Food industries: A Review.
L-glutaminase has versatile applications in pharma and food industries. In pharmaceutical industry, L-glutaminase can be used as anti-oxidant and anti-cancer agent to treat Acute Lymphocytic Leukaemia (ALL). Whereas, in the food industry, L-glutaminase is used for acrylamide degradation, theanine production, flavour enhancer, soy sauce and many. The other applications include nitrogen metabolism and its use as biosensor in hybridoma technology. Both intra-cellular and extra-cellular L-glutaminases from wide range of sources were identified. Because of its diverse applications, there is a need to improve the production of L-glutaminase by enzyme engineering technology. Effect of recombination on L-glutaminase production was also reported. Researchers also confirmed the antitumor properties of L-glutaminase by conducting in vitro, in vivo and in silico studies. Bacillus sps. and Aspergillus sps. are the commercial producers of L-glutaminase. In this review, the applications, different sources of Lglutaminase, anti-cancer properties were discussed.


Acute chest pain in contingency operations at a Role 1 facility.
Acute chest pain is a common medical presenting complaint which can be difficult to diagnose and treat outside of a fully equipped emergency department. In future contingency operations the number of personnel deployed is likely be smaller, with the medical cover appropriate for the population at risk, such that the deployed medical facilities will be smaller than the Role 3 unit with which we have become familiar over the last 10 years of operations in Afghanistan. Physician involvement in these smaller medical facilities is crucial to maintain clinical effect when dealing with patients presenting with disease and non-battle injury, which can often make up the majority of deployed healthcare work. Patients presenting with chest pain require rapid assessment and stabilisation prior to medical evacuation to a suitable definitive care unit. This article focuses on emergency acute chest pain presentations, non-cardiac causes of chest pain, risk reduction and how contingency will affect patient care.


Expression of MSH2 and MSH6 on a tissue microarray in patients with osteosarcoma.
Reliable prognostic factors for the outcome of patients with osteosarcoma (OS) remain elusive. We analyzed the relationship between immunohistochemical expression of deoxyribonucleic acid (DNA) mismatch repair (MMR) proteins, MutS protein homolog 2 (MSH2) and MSH6 using a tissue microarray (TMA) with respect to OS patient demographics and survival time. We retrieved tumor tissue specimens from bone tissue originating from surgical primary tumor specimens of OS patients to generate a TMA and stained sections with antibodies against MSH2 and MSH6. Tumor resections of 67 patients with a mean follow-up of 98 months were evaluated. MSH2 was expressed in nine (13%), MSH6 in ten (15%) and combined MSH2 and MSH6 (MSH2/6) in six (9%) patients. Significantly shorter survival times were associated with expression of MSH6, MSH2/6, as well as simultaneous non-response to chemotherapy and presence of metastasis. The survival time of patients with OS may be predicted by local expression of MSH6 and MSH2/6 in surgical primary tumor resections. This study shows the prognostic value of the local expression of DNA MMR proteins, as markers for poor prognosis of OS patients.


Southern Africa crustal anisotropy reveals coupled crust-mantle evolution for over 2 billion years.
The long-term stability of Precambrian continental lithosphere depends on the rheology of the lithospheric mantle as well as the coupling between crust and mantle lithosphere, which may be inferred by seismic anisotropy. Anisotropy has never been detected in cratonic crust. Anisotropy in southern Africa, detected by the seismological SKS-splitting method, usually is attributed to the mantle due to asthenospheric flow or frozen-in features of the lithosphere. However, SKS-splitting cannot distinguish between anisotropy in the crust and the mantle. We observe strong seismic anisotropy in the crust of southern African cratons by Receiver Function analysis. Fast axes are uniform within tectonic units and parallel to SKS axes, orogenic strike in the Limpopo and Cape fold belts, and the strike of major dyke swarms. Parallel fast axes in the crust and mantle indicate coupled crust-mantle evolution for more than 2 billion years with implications for strong rheology of the lithosphere.


Pacific and Atlantic Lepeophtheirus salmonis (Krøyer, 1838) are allopatric subspecies: Lepeophtheirus salmonis salmonis and L. salmonis oncorhynchi subspecies novo.
The salmon louse Lepeophtheirus salmonis is a parasitic copepod that infects salmonids in the Pacific and Atlantic oceans. Although considered as a single species, morphological and biological differences have been reported between lice from the two oceans. Likewise, studies based on nucleotide sequencing have demonstrated that sequence differences between Atlantic and Pacific L. salmonis are highly significant, albeit smaller than the divergence observed between congeneric copepod species. We demonstrated reproductive compatibility between L. salmonis from the two oceans and successfully established F2 hybrid strains using separate maternal lines from both the Pacific and Atlantic. The infection success for the F2 hybrid strains were similar to results typically observed for non hybrid lice strains in the rearing facility used. Lepeophtheirus salmonis COI and 16S sequences divergence between individuals from the Pacific and the Atlantic oceans was high compared to what may be expected within a copepod species and phylogenetic analysis showed that they consistently formed monophyletic clades representing their origin from the Pacific or Atlantic oceans. Lepeophtheirus salmonis from the Pacific and Atlantic oceans are reproductively compatible at least until adults at the F2 hybrid stage, and should not be regarded as separate species based on reproductive segregation or sequence divergence levels. Reported biological and genetic differences in L. salmonis seen in conjunction with the reported genetic diversity commonly observed between and within species demonstrate that Atlantic and Pacific L. salmonis should be regarded as two subspecies: Lepeophtheirus salmonis salmonis and L. salmonis oncorhynchi subsp. nov.


The spatial variability of soil resources following long-term disturbance.
The spatial distributions of selected soil properties in two adjacent sites in southwest Michigan were examined to evaluate the potential effects of chronic disturbance on resource heterogeneity. One site was a cultivated field that had been cleared, plowed, and cropped annually for decades prior to sampling while the other, uncultivated field was cleared of original forest in 1960 after which it was mown annually but never plowed or cropped. We took replicate samples from a 330-point unaligned grid across the sites for soil pH, gravimetric moisture, inorganic phosphorus, total carbon, and net nitrification and nitrogen mineralization potentials. Soils in the cultivated site contained less than half as much carbon as in the uncultivated site, but had higher levels of inorganic phosphorus and moisture, and higher soil pH. Potential net nitrogen mineralization and nitrification rates did not differ between sites. Geostatistical analysis showed that almost all properties examined were strongly autocorelated within each site; structural variance as a proportion of sample variance ranged from 30-95% for all properties, and for any given property differed little between sites. The distance over which this dependence was expressed, however, was for all properties but pH substantially less in the uncultivated site (7-26 m) as compared to the tilled site (48-108m), especially for total C and net nitrification and N mineralization. These results suggest that the spatial pattern and scale of soil variability can differ markedly among edaphically identical sites and that these differences can be related to disturbance history.


In planta conditions induce genomic changes in Pseudomonas syringae pv. phaseolicola.
The co-evolution of bacterial plant pathogens and their hosts is a complex and dynamic process. Plant resistance can impose stress on invading pathogens that can lead to, and select for, beneficial changes in the bacterial genome. The Pseudomonas syringae pv. phaseolicola (Pph) genomic island PPHGI-1 carries an effector gene, avrPphB (hopAR1), which triggers the hypersensitive reaction in bean plants carrying the R3 resistance gene. Interaction between avrPphB and R3 generates an antimicrobial environment within the plant, resulting in the excision of PPHGI-1 and its loss from the genome. The loss of PPHGI-1 leads to the generation of a Pph strain able to cause disease in the plant. In this study, we observed that lower bacterial densities inoculated into resistant bean (Phaseolus vulgaris) plants resulted in quicker PPHGI-1 loss from the population, and that loss of the island was strongly influenced by the type of plant resistance encountered by the bacteria. In addition, we found that a number of changes occurred in the bacterial genome during growth in the plant, whether or not PPHGI-1 was lost. We also present evidence that the circular PPHGI-1 episome is able to replicate autonomously when excised from the genome. These results shed more light onto the plasticity of the bacterial genome as it is influenced by in planta conditions.


Involvement of Ahr Pathway in Toxicity of Aflatoxins and Other Mycotoxins.
The purpose of this review is to present information about the role of activation of aflatoxins and other mycotoxins, of the aryl hydrocarbon receptor (AhR) pathway. Aflatoxins and other mycotoxins are a diverse group of secondary metabolites that can be contaminants in a broad range of agricultural products and feeds. Some species of Aspergillus, Alternaria, Penicilium, and Fusarium are major producers of mycotoxins, some of which are toxic and carcinogenic. Several aflatoxins are planar molecules that can activate the AhR. AhR participates in the detoxification of several xenobiotic substances and activates phase I and phase II detoxification pathways. But it is important to recognize that AhR activation also affects differentiation, cell adhesion, proliferation, and immune response among others. Any examination of the effects of aflatoxins and other toxins that act as activators to AhR must consider the potential of the disruption of several cellular functions in order to extend the perception thus far about the toxic and carcinogenic effects of these toxins. There have been no Reviews of existing data between the relation of AhR and aflatoxins and this one attempts to give information precisely about this dichotomy.


Inferred spoligoforest topology unravels spatially bimodal distribution of mutations in the DR region.
Biomarkers of Mycobacterium tuberculosis complex (MTBC) mutate over time. Among the biomarkers of MTBC, spacer oligonucleotide type (spoligotype) and mycobacterium interspersed repetitive unit (MIRU) patterns are commonly used to genotype clinical MTBC strains. In this study, we present an evolution model of spoligotype rearrangements using MIRU patterns to disambiguate the ancestors of spoligotypes. We use a large patient dataset from the United States Centers for Disease Control and Prevention (CDC) to generate this model. Based on the contiguous deletion assumption and rare observation of convergent evolution, we first generate the most parsimonious forest of spoligotypes, called a spoligoforest, using three genetic distance measures. An analysis of topological attributes of the spoligoforest and number of variations at the direct repeat (DR) locus of each strain reveals interesting properties of deletions in the DR region. First, we compare our mutation model to existing mutation models of spoligotypes and find that our mutation model produces as many within-lineage mutation events as other models, with slightly higher segregation accuracy. Second, based on our mutation model, the number of descendant spoligotypes follows a power law distribution. Third, contrary to prior studies, the power law distribution does not plausibly fit to the mutation length frequency. Moreover, we find that the total number of mutation events at consecutive spacers follows a spatially bimodal distribution. The two modes are spacers 13 and 40, which are hotspots for chromosomal rearrangements, and the change point is spacer 34, which is absent in most MTBC strains. Based on this observation, we built two alternative models for mutation length frequency: the Starting Point Model (SPM) and the Longest Block Model (LBM). Both models are plausibly good fits to the mutation length frequency distribution, as verified by the goodness-of-fit test. We also apply SPM and LBM to a dataset from Institut Pasteur de Guadeloupe and verify that these models hold for different strain datasets.


Peritoneal transport status correlates with morbidity but not longitudinal change of nutritional status of continuous ambulatory peritoneal dialysis patients: a 2-year prospective study.
Cross-sectional studies show that peritoneal transport status is associated with nutritional status and clinical outcome of continuous ambulatory peritoneal dialysis (CAPD) patients. High transporters often are assumed to have progressive malnutrition as a result of fluid overload and nutrient loss in dialysis effluent. There are few prospective data to confirm this assumption, however. We studied 235 unselected CAPD patients to examine this possibility. There were 58 new cases and 177 prevalent cases. A standard peritoneal equilibration test was performed at enrollment. All patients were followed for 2 years. Dialysis adequacy and nutritional assessment, including serum albumin, protein nitrogen appearance, and lean body mass, were performed at 0, 12, and 24 months. Clinical outcome included hospitalization, actuarial patient survival, and technique survival. The mean age was 51.6 +/- 12.4 years. The mean 4-hour dialysate-to-plasma ratio of creatinine was 0.57 +/- 0.12 for all cases (0.60 +/- 0.12 for new cases). Patients were classified into three groups: high/high-average (H/HA) (63 cases), low-average (LA) (105 cases), and low (L) (67 cases) transporters. Baseline demographic data and total Kt/V were similar in all transport groups. There were weak correlations between 4-hour dialysate-to-plasma ratio of creatinine and baseline serum albumin (r = -0.249, P: < 0.001), protein nitrogen appearance (r = -0.190, P: < 0.01), and percentage of lean body mass (r = -0.194, P: < 0.01). The H/HA group was a specific but not a sensitive predictor of poor baseline nutritional status. There was no significant change in any nutritional indices after 2 years in new cases and prevalent cases, regardless of transport status. The differences in nutritional indices between groups remained unchanged during the study period. There was a significant difference in hospitalization rate among peritoneal transport groups (median 12 versus 7 versus 3 days per year for H/HA, LA, and L groups, Kruskal-Wallis test, P: < 0.05). The difference remained similar when new cases and prevalent cases were analyzed separately. The L group had slightly better 2-year patient survival than the H/HA group (90.2% versus 83.3%), but the result was not statistically significant. We conclude that peritoneal transport status is not associated with longitudinal change of nutritional parameters, although transport status is associated with short-term patient morbidity. Further study is needed to identify the mechanisms of poor clinical outcome in high peritoneal transporters.


Distribution of heart rate and responses to resuscitation among 1237 apnoeic newborns at birth.
Describe the distribution of the first recorded heart rate (HR) in apnoeic term/near-term newborns, HR responses to basic resuscitation (no intubation, chest compressions and/or medication), and relationship to 24-h outcomes. We also document patient characteristics and care provider behaviour stratified by first HR. Descriptive study from July 2013 through June 2018 at Haydom Hospital in Tanzania. All deliveries were observed by assistants recording data. Bag-mask ventilation and ECG data were recorded by resuscitation monitors. Newborns with ≥5 ventilations and ECG signal-data were included. 1237 term/near-term newborns with median (25th, 75th percentiles) gestation 38 (37, 40) weeks and birth weight 3140 (2750, 3500) grams fulfilled inclusion criteria. The first HR, measured median 102 (73, 144) s after birth following drying/stimulation, was distributed into two peaks with centres around 60 and 165 bpm, 51% were ≥100 bpm. After ventilation, the HR distribution shifted to a single-peak, with median 161 bpm. At least one low-high HR transition crossing 100 bpm was noted in 44% of newborns. The HR increase occurred over median 9.2 (6.2, 13) s, was 60 (43, 77) bpm, and 86% followed a ventilation sequence of 23 (16, 34) s duration. 72% of the newborns with first HR < 60 bpm survived following ventilations only. Both first and final HR were significantly related to 24-h outcomes. The first recorded HR was distributed into two peaks on each side of 100 bpm. Ventilation increased HR in most newborns. Lower first and final HR were related to gradually more adverse 24-h outcomes.


Performance Characteristics of Next-Generation Sequencing for the Detection of Antimicrobial Resistance Determinants in Escherichia coli Genomes and Metagenomes.
Short-read sequencing can provide detection of multiple genomic determinants of antimicrobial resistance from single bacterial genomes and metagenomic samples. Despite its increasing application in human, animal, and environmental microbiology, including human clinical trials, the performance of short-read Illumina sequencing for antimicrobial resistance gene (ARG) detection, including resistance-conferring single nucleotide polymorphisms (SNPs), has not been systematically characterized. Using paired-end 2 × 150 bp (base pair) Illumina sequencing and an assembly-based method for ARG prediction, we determined sensitivity, positive predictive value (PPV), and sequencing depths required for ARG detection in an Escherichia coli isolate of sequence type (ST) 38 spiked into a synthetic microbial community at varying abundances. Approximately 300,000 reads or 15× genome coverage was sufficient to detect ARGs in E. coli ST38, with comparable sensitivity and PPV to ~100× genome coverage. Using metagenome assembly of mixed microbial communities, ARG detection at E. coli relative abundances of 1% would require assembly of approximately 30 million reads to achieve 15× target coverage. The minimum sequencing depths were validated using public data sets of 948 E. coli genomes and 10 metagenomic rectal swab samples. A read-based approach using k-mer alignment (KMA) for ARG prediction did not substantially improve minimum sequencing depths for ARG detection compared to assembly of the E. coli ST38 genome or the combined metagenomic samples. Analysis of sequencing depths from recent studies assessing ARG content in metagenomic samples demonstrated that sequencing depths had a median estimated detection frequency of 84% (interquartile range: 30%-92%) for a relative abundance of 1%. IMPORTANCE Systematically determining Illumina sequencing performance characteristics for detection of ARGs in metagenomic samples is essential to inform study design and appraisal of human, animal, and environmental metagenomic antimicrobial resistance studies. In this study, we quantified the performance characteristics of ARG detection in E. coli genomes and metagenomes and established a benchmark of ~15× coverage for ARG detection for E. coli in metagenomes. We demonstrate that for low relative abundances, sequencing depths of ~30 million reads or more may be required for adequate sensitivity for many applications.


Pediatric Forefoot Fractures: Assessment of Fracture Patterns and Predictors of Complicated Outcome.
Forefoot fractures account for 6% to 10% of fractures in children, and although the majority heals with supportive treatment, complications may lead to pain and disability. No previous study in children has evaluated complication risk in the emergency department based on initial assessment characteristics. The study aim was to identify the radiological and clinical variables that increase the complication rate of pediatric forefoot fractures. This may help emergency physicians refer patients who require more thorough follow-up or surgical intervention. We evaluated 497 forefoot fractures on initial presentation to a pediatric emergency department at the Children's Hospital at London Health Science Centre over a 6-year period. We collected variables such as degree of angulation, displacement, number of concurrent fractures, and demographic data such as age and sex. We then determined the variables associated with complications by reviewing each patient's chart. Overall, there was a 6.4% complication rate. Analysis identified sex as an important predictor of complications. Females, although representing approximately one third of the sample, represented nearly two thirds of the cases with complicated outcomes (P = 0.001; odds ratio [OR], 4.67). Increased number of fractures was also significant (P = 0.01; OR, 2.41) as was increasing age (P = 0.01; OR, 1.17) and patients who chose to return to the emergency department (P < 0.05; OR, 5.282). Lateral angulation/displacement and anteroposterior angulation/displacement were not associated with increased complications. Identifying features, such as female sex, increasing age, multiple fractures, and return to emergency departments for repeat visits, may help guide the emergency physician on whom to refer for specialized care.


A roadmap to disentangle the molecular etiology of schizophrenia.
Schizophrenia is a severe mental disorder striking mainly young adults and leading to life-long disability in a substantial portion of the sufferers. On the other hand, substantial knowledge about its etiology and pathogenesis is still lacking. Therefore the European Science Foundation (ESF) sponsored a meeting of a panel of European experts on schizophrenia research to discuss the state of art and future perspectives of key topics in this area. The fields covered genetics, epidemiology, animal models, molecular neuropathology and imaging. This was a first step to establish a network of European groups dedicated to Schizophrenia research. The coming calls of the frame work program will be used to strengthen this network in order to achieve substantial progress in understanding and treating this devastating illness.


Development and function of human dendritic cells in humanized mice models.
Dendritic cells (DCs) are sentinel cells of the immune system arising from hematopoietic stem cells. DCs play a key role in the regulation of both adaptive and innate lymphocyte responses. As such, experimental models enabling a thorough analysis of human DCs development and function are needed. Humanized mice models (termed collectively as HIS mice, or human immune system mice models) provide unique opportunities to model human hematopoiesis and tackle the function of human immune cell types in vivo. Here, we review experimental approaches enabling to recapitulate the ontogeny of DC subsets in HIS mice and discuss studies addressing the biology of human DC subsets implementing HIS mice models.


Neuroblastoma of undifferentiated subtype, prognostic significance of prominent nucleolar formation, and MYC/MYCN protein expression: a report from the Children's Oncology Group.
This study sought to investigate biological/clinicopathological characteristics of neuroblastoma, undifferentiated subtype (NBUD). This study examined 157 NBUD cases filed at the Children's Oncology Group Neuroblastoma Pathology Reference Laboratory, and survival rates of the patients were analyzed with known prognostic factors. Immunostainings for MYCN and MYC protein were performed on 68 tumors. NBUD cases had a poor prognosis (48.4% ± 5.0% 3-year event-free survival [EFS]; 56.5% ± 5.0% overall survival [OS]), and were often associated with high mitosis-karyorrhexis index (MKI, 65%), prominent nucleoli (PN, 83%), ≥ 18 months of age (75%), MYCN amplification (MYCN-A, 83%), diploid pattern (63%), and 1pLOH (loss of heterozygosity (72%). However, these prognostic indicators, except for MYCN status, had no significant impact on survival. Surprisingly, EFS for patients with MYCN-A tumors (53.4% ± 5.6%) was significantly better (P=.0248) than for patients with MYCN-nonamplified (MYCN-NA) tumors (31.7% ± 11.7%), with MYCN-NA and PN (+) tumors having the worst prognosis (9.3% ± 8.8%, P=.0045). Immunohistochemically, MYCN expression was found in 42 of 48 MYCN-A tumors. In contrast, MYC expression was almost exclusively found in the MYCN-NA tumors (9 of 20) especially when they had PN (8 of 11). Those patients with only MYC-positive tumors had the worst EFS (N=8, 12.5% ± 11.7%) compared with only MYCN-positive (N=39, 49.9% ± 17.7%) and both negative tumors (N=15, 70.0% ± 17.1%) (P= .0029). High MKI was often found in only MYCN-positive (30 of 38) but rarely in only MYC-positive (2 of 8) tumors. NBUD represents a unique subtype of neuroblastoma associated with a poor prognosis. In this subtype, MYC protein expression may be a new prognostic factor indicating more aggressive clinical behavior than MYCN amplification and subsequent MYCN protein expression.


Improved human tenocyte proliferation and differentiation in vitro by optimized silk degumming.
Tendon disorders are common clinical conditions. Tendon tissue engineering provides a new approach for tendon repair by integrating engineered substitutes with their native counterparts. Silk is considered to be a promising candidate for tendon engineering because of its biological and mechanical properties. However, a major concern with using silk for biomedical applications is the immune responses generated by sericin, a glue-like protein that coats the silk fibres. This study improves the existing protocols for silk 'degumming' which removes sericin and enables preparation of silk that is suitable for tendon regeneration. Bombyx mori silks were treated by sequential treatments with different proteases. The efficiency of degumming was determined by measuring weight loss, picric acid and carmine staining and scanning electron microscopy. To evaluate the cellular responses after degumming, the growth and differentiation of human tenocytes on silks were examined. The results showed that sequential protease treatment effectively degummed raw silks. The sequentially degummed silks showed enhanced tenocyte proliferation and upregulated mRNA levels of tendon markers. Thick cell multilayers formed on the treated silks, with cells and collagen fibres penetrating into the spaces in individual silk filaments, resulting in a structure resembling human tendon.


Baroreflex sensitivity and outcomes following coronary surgery.
Postoperative atrial fibrillation, acute kidney dysfunction and low cardiac output following coronary surgery are associated with morbidity and mortality. The purpose of this study is to determine if the preoperative autonomic control is a determinant of these postoperative complications. This is a prospective cohort study on 150 adult patients undergoing surgical coronary revascularization with cardiopulmonary bypass. The patients received an autonomic control assessment after the induction of anesthesia. Baroreflex sensitivity was computed by spectral analysis and expressed as BRSαHF and BRSαLF for measure respectively in the high and low frequency domains. Atrial fibrillation was adjudicated at any postoperative time during the hospital stay. Acute kidney dysfunction was defined as any increase of serum creatinine levels from preoperative values within the first 48 hours after surgery, and acute kidney injury was adjudicated at a 50% increase. Low cardiac ouput syndrome was defined as the need for inotropic support > 48 hours. Thirty-eight (26.4%) patients experienced postoperative atrial fibrillation; 32 (22.2%) had acute kidney dysfunction and 5 (3.5%) acute kidney injury; 14(10%) had a low cardiac output state. No indices of baroreflex sensitivity were associated with atrial fibrillation or acute kidney injury. A low value of BRSαLF was associated with acute kidney dysfunction and low cardiac output state. A BRSαLF < 3 msec/mmHg was an independent risk factor for acute kidney dysfunction (odds ratio 3.0, 95% confidence interval 1.02-8.8, P = 0.045) and of low cardiac output state (odds ratio 17.0, 95% confidence interval 2.9-99, P = 0.002). Preoperative baroreflex sensitivity is linked to postoperative complications through a number of possible mechanisms, including an autonomic nervous system-mediated vasoconstriction, a poor response to hypotension, and an increased inflammatory reaction.