Alliin alters gut microbiota and gene expression of colonic epithelial tissues.
Alliin is a natural organosulfur-containing phytochemical in garlic. It is possible that alliin can regulate the gut microbiota for its strong antimicrobial activity against many pathogens. Here, we assessed whether alliin impacts the distal small intestinal bacteria, hence the cecal microbiota, thus altering the gene expression of colonic epithelial tissues (CETs). Eighty mg/kg alliin was orally administered to rats for 14 days, and the 16S rDNA from small intestinal and cecal microbiota as well as mRNA from CETs were sequenced and analyzed. The results showed that alliin consumption affected microbiota composition in both the small intestine and cecum, although there was only one specific genus, Allobaculum that was significantly altered in the rat cecum. The altered composition of microbiota indirectly impacted 174 genes in the CETs. Specifically, five genes, including RT1-Ba, RT1-Bb, Cd80, Madcam1, and Aicda, indicated this consumption related to the intestinal immune network for IgA production. PRACTICAL APPLICATIONS: We firstly reported alliin consumption in vivo potentially affected the intestinal immunity of healthy rats by slightly alteration of microbiota composition in small intestine and cecum. The alteration subsequently amplified, resulting in the change of the colonic epithelial expression of several genes related to the intestinal immune network for IgA production. Hence, we suggested the alliin consumption may potentially affect the immune system of healthy individuals by alteration of gut microbiota and epithelial gene expression.


Characteristics of general and subspecialty pediatricians who choose not to recertify.
The goal was to determine the reasons given for nonparticipation in maintenance of certification by general pediatricians and pediatric subspecialists whose board certification had expired. All United States-based pediatricians whose board certification had expired and who had not recertified as a generalist or subspecialist in 2004 or 2005 were surveyed. The total sample included 1001 pediatric generalists and 1237 subspecialists. We conducted a mail survey study, with a total of 3 mailings to nonrespondents. Analyses initially were conducted separately for generalists with expired certificates, subspecialists with active subspecialty but expired general pediatrics certificates, and subspecialists with expired subspecialty certificates (regardless of their general pediatrics certification status). The overall response rates were 68% for generalists and 76% for subspecialists. Of pediatricians who had allowed their certificates to expire, the majority still intended to recertify (65% of the generalists and 86% of the subspecialists, in their primary subspecialty). The most common reasons cited by the 35% of generalists with expired certificates who were not planning to participate in maintenance of certification were the expense, the time required to complete maintenance of certification, and the perceived lack of relevance to their current practice. The 14% of subspecialists who were not planning to recertify in their primary subspecialty most commonly cited the expense, a change in career path making recertification unnecessary, and the time required. Although the physicians in this study had allowed their certificates to expire, they still overwhelmingly believed that physicians who provide direct patient care should maintain their certification. There was general consensus among our respondents regarding the professional value to patients and peers of participation in the maintenance of certification program.


Impact of transfusions and acute rejection on posttransplantation donor antigen-specific responses in two study populations. Cooperative Clinical Trial in Transplantation Research Group.
We participated in a protocol supported by the National Institutes of Allergy and Infectious Disease, Cooperative Clinical Trial in Transplantation (CCTT), which was designed to investigate the effect of peritransplant donor-specific transfusion in non-HLA-identical living donor kidney recipients. We determined the donor antigen-specific responses at 1 year after transplantation for the 79 CCTT donor-recipient combinations in this study. A lower rate of donor antigen-specific hyporeactivity was seen in the CCTT recipients (6 of 79=8%) versus our recipients at the University of Minnesota who underwent transplantation in the same period (9 of 55=16%, P=0.16) and versus our combined historical data (33 of 131=25%, P=0.002). Therefore, we studied the differences in the two recipient populations to determine why hyporeactivity was lower in the CCTT group than at our center. Significant differences were seen in the acute rejection rates and the frequency of pretransplantation random transfusion. Overall and early (<3 month) acute rejection rates were higher in CCTT versus Minnesota recipients (overall: 51% vs. 20%, P=0.001) (early: 43% vs. 16%, P=0.001). The frequency of pretransplantation random transfusion was 40% for CCTT recipients (34%) versus 80% for Minnesota recipients (75%) (P=0.0004). These results provide provocative, although not conclusive, evidence for the importance of pretransplantation transfusion and acute rejection episodes in the development of donor antigen-specific hyporeactivity. Pre-, peri-, and posttransplantation clinical events undoubtedly have an impact on posttransplantation immune parameters.


Asserting Our Competence and Affirming the Value of Occupation With Confidence.
Throughout the years, leaders in the profession have challenged us to affirm the value of occupational therapy and to substantiate what we do. Occupational therapy practitioners have always focused on what most matters to clients in what is now called client-centered or patient-centered practice. We have also focused on client function to enable participation in everyday life. In a welcome shift, society's views about health and meaning-making are becoming more congruent with the long-standing ideals of occupational therapy. Now, more than ever, we have a powerful opportunity to communicate our competence. But how do we assert our competence and the complexity of occupation with confidence? This lecture draws on the conceptual foundations of theories about competence and confidence and provides examples from the research literature, and a practitioner and client narrative to illustrate the factors that enable us to effectively demonstrate the value of occupational therapy.


Archaeal DNA Polymerase-B as a DNA Template Guardian: Links between Polymerases and Base/Alternative Excision Repair Enzymes in Handling the Deaminated Bases Uracil and Hypoxanthine.
In Archaea repair of uracil and hypoxanthine, which arise by deamination of cytosine and adenine, respectively, is initiated by three enzymes: Uracil-DNA-glycosylase (UDG, which recognises uracil); Endonuclease V (EndoV, which recognises hypoxanthine); and Endonuclease Q (EndoQ), (which recognises both uracil and hypoxanthine). Two archaeal DNA polymerases, Pol-B and Pol-D, are inhibited by deaminated bases in template strands, a feature unique to this domain. Thus the three repair enzymes and the two polymerases show overlapping specificity for uracil and hypoxanthine. Here it is demonstrated that binding of Pol-D to primer-templates containing deaminated bases inhibits the activity of UDG, EndoV, and EndoQ. Similarly Pol-B almost completely turns off EndoQ, extending earlier work that demonstrated that Pol-B reduces catalysis by UDG and EndoV. Pol-B was observed to be a more potent inhibitor of the enzymes compared to Pol-D. Although Pol-D is directly inhibited by template strand uracil, the presence of Pol-B further suppresses any residual activity of Pol-D, to near-zero levels. The results are compatible with Pol-D acting as the replicative polymerase and Pol-B functioning primarily as a guardian preventing deaminated base-induced DNA mutations.


End of life of patients treated with haemodialysis as narrated by their close relatives.
The study aimed to describe end of life for patients treated with maintenance haemodialysis as narrated by their close relatives. Many patients undergoing haemodialysis are older, have several comorbidities and underestimated symptoms and are in their last year of life. To improve care, we need to know more about their end-of-life situation. Qualitative and descriptive. Qualitative retrospective interviews were conducted with 14 close relatives of deceased haemodialysis patients (3-13 months after death). Data were analysed using qualitative content analysis. The study is ethically approved. In the last months, a gradual deterioration in health with acute episodes necessitating hospital admissions was described. This involved diminishing living space and expressions of dejection, but also of joy. Three patterns emerged in the last weeks: uncertain anticipation of death as life fades away; awaiting death after haemodialysis withdrawal; and sudden but not unexpected death following intensive care. Findings show complexities of decisions on haemodialysis withdrawal. Different end-of-life patterns all involved increasingly complex care needs and existential issues. Findings show a need for earlier care planning. The identification of organisational factors to facilitate continuity and whole person care to meet these patients' specific care needs with their complex symptom burdens and comorbidities is needed. Findings indicate the need for integration of a palliative care approach in the treatment of patients in haemodialysis care.


HVint: A Strategy for Identifying Novel Protein-Protein Interactions in Herpes Simplex Virus Type 1.
Human herpesviruses are widespread human pathogens with a remarkable impact on worldwide public health. Despite intense decades of research, the molecular details in many aspects of their function remain to be fully characterized. To unravel the details of how these viruses operate, a thorough understanding of the relationships between the involved components is key. Here, we present HVint, a novel protein-protein intraviral interaction resource for herpes simplex virus type 1 (HSV-1) integrating data from five external sources. To assess each interaction, we used a scoring scheme that takes into consideration aspects such as the type of detection method and the number of lines of evidence. The coverage of the initial interactome was further increased using evolutionary information, by importing interactions reported for other human herpesviruses. These latter interactions constitute, therefore, computational predictions for potential novel interactions in HSV-1. An independent experimental analysis was performed to confirm a subset of our predicted interactions. This subset covers proteins that contribute to nuclear egress and primary envelopment events, including VP26, pUL31, pUL40, and the recently characterized pUL32 and pUL21. Our findings support a coordinated crosstalk between VP26 and proteins such as pUL31, pUS9, and the CSVC complex, contributing to the development of a model describing the nuclear egress and primary envelopment pathways of newly synthesized HSV-1 capsids. The results are also consistent with recent findings on the involvement of pUL32 in capsid maturation and early tegumentation events. Further, they open the door to new hypotheses on virus-specific regulators of pUS9-dependent transport. To make this repository of interactions readily accessible for the scientific community, we also developed a user-friendly and interactive web interface. Our approach demonstrates the power of computational predictions to assist in the design of targeted experiments for the discovery of novel protein-protein interactions.


Whole genome sequencing analysis of the cutaneous pathogenic yeast Malassezia restricta and identification of the major lipase expressed on the scalp of patients with dandruff.
Malassezia species are opportunistic pathogenic fungi that are frequently associated with seborrhoeic dermatitis, including dandruff. Most Malassezia species are lipid dependent, a property that is compensated by breaking down host sebum into fatty acids by lipases. In this study, we aimed to sequence and analyse the whole genome of Malassezia restricta KCTC 27527, a clinical isolate from a Korean patient with severe dandruff, to search for lipase orthologues and identify the lipase that is the most frequently expressed on the scalp of patients with dandruff. The genome of M. restricta KCTC 27527 was sequenced using the Illumina MiSeq and PacBio platforms. Lipase orthologues were identified by comparison with known lipase genes in the genomes of Malassezia globosa and Malassezia sympodialis. The expression of the identified lipase genes was directly evaluated in swab samples from the scalps of 56 patients with dandruff. We found that, among the identified lipase-encoding genes, the gene encoding lipase homolog MRES_03670, named LIP5 in this study, was the most frequently expressed lipase in the swab samples. Our study provides an overview of the genome of a clinical isolate of M. restricta and fundamental information for elucidating the role of lipases during fungus-host interaction.


Neurocysticercosis in Houston, Texas: an update.
Neurocysticercosis, one of the most common parasitic infections of the human nervous system, has emerged as an important infection in the United States. Neurocysticercosis causes significant morbidity associated with acute seizures, chronic epilepsy, and hydrocephalus.We retrospectively identified patients with definitive or probable neurocysticercosis seen at Ben Taub General Hospital, the largest public teaching hospital in Houston, Texas, from September 1997 through December 2005. We collected demographic, clinical, therapeutic, and outcome variables. Neurocysticercosis was classified according to the location of cysts in imaging studies. We compared cases with parenchymal and extraparenchymal disease.We included 111 patients (48 had definitive and 63 probable neurocysticercosis). The mean age was 28.6 years (standard deviation, 13.6 yr), and the male to female ratio was 2:1. Most patients (93%) were Hispanic immigrants. Sixty (54%) patients had parenchymal disease, 22 (20%) intraventricular, 13 (12%) subarachnoid disease, and 13 (12%) had calcifications only. Additionally, 2 patients had hydrocephalus only, and 1 had ocular cysticercosis. Thirteen (40%) of 32 patients with parenchymal disease and 3 (30%) of 10 patients with calcifications had relapsed seizures at follow-up. Extraparenchymal disease was associated with longer duration of hospitalization compared with parenchymal disease. No deaths were identified in our series during a median follow-up of 1 year.Neurocysticercosis has emerged as an important parasitic infection in developed countries as a result of increased migration. With current management, mortality is limited, but there continues to be significant morbidity. Further studies of the epidemiology and pathophysiology of the infection are urgently needed to develop better preventive and therapeutic strategies.


Dose-response analysis in risk assessment: evaluation of behavioral specificity.
Several methods of quantitative risk assessment that have been described recently are particularly applicable to neurotoxic end points. These methods can be broadly divided into two types of approaches based on their treatment of dose-response data to estimate risks. Benchmark approaches estimate risks using variability in response to a fixed dose level in comparison with background control variability. Probabilistic approaches estimate risks using the variability in the dose to produce a small effect in the sample population. The current report seeks to extend the development of probabilistic approaches for neurotoxic end points. Because behavioral data are often used to assess therapeutic efficacy as well as toxicity (unwanted effects), this analysis focused on the relative risks of producing these effects with the same agent. The therapeutic potential of GBR 12909 was determined by its ability to decrease cocaine-maintained responding in monkeys. The effects of this agent were also assessed in the same monkeys using food-maintained responding to provide an indication of behavioral toxicity. GBR 12909 decreased both behaviors, with complete decreases on drug-seeking behavior occurring at doses that had minimal effects on food-maintained responding. The difference in the estimates of doses to decrease drug-seeking and food-maintained behavior suggested that specific therapeutic effects could be obtained in the absence of unwanted side effects for a definable proportion of the population. These results also suggest that multiple behavioral end points can be useful for identifying specific effects of chemicals for the purposes' of risk assessment.


Molecular identification and expression of differentially regulated genes of the European flounder, Platichthys flesus, submitted to pesticide exposure.
Effects of pesticide exposure on the European flounder, Platichthys flesus, were investigated using a suppression subtractive hybridization method (SSH) to identify up- and down-regulated genes after a 30-day exposure to herbicides (a cocktail of atrazine, diuron, and isoproturon, and a single herbicide, glyphosate). A total of 256 expressed gene sequences were identified as having the potential for being differentially expressed, of which 116 could be identified by homology with databased sequences. The metabolic functions with which they are associated include energy production, general metabolism, signaling, transport, immune system, and structure. Expression of 14 of these genes was analyzed in liver, muscle, and gills by reverse transcriptase-polymerase chain reaction (RT-PCR) under experimental conditions (0, 15, and 30 days of exposure) and under field conditions (sampling in two estuaries displaying different levels of pesticide contamination). This study provides a first basis for studying the response of fish to pesticide exposure and allows the characterization of new potential genetic markers of pesticide contamination in the field.


CaMKII inhibition with KN93 attenuates endothelin and serotonin receptor-mediated vasoconstriction and prevents subarachnoid hemorrhage-induced deficits in sensorimotor function.
It has been suggested that transcriptional upregulation of cerebral artery contractile endothelin (ETB) and 5-hydroxytryptamine (5-HT1B) receptors play an important role in the development of late cerebral ischemia and increased vasoconstriction after subarachnoid hemorrhage (SAH). We tested the hypothesis that inhibition of calcium calmodulin-dependent protein kinase II (CaMKII) may reduce cerebral vasoconstriction mediated by endothelin and serotonin receptors and improve neurological outcome after experimental SAH. SAH was induced in adult rats by injection of 250 μL autologous blood into the basal cisterns. The CaMKII activity in cerebral vessels was studied by Western blot and immunohistochemistry. The vasomotor responses of middle cerebral and basilar arteries were measured in a sensitive myograph system. The functional outcome was examined by the rotating pole test 2 and 3 days after SAH. SAH induced a rapid early increase in phosphorylated CaMKII protein at 1 h that was attenuated by cisternal administration of the CaMKII inhibitor KN93 (0.501 μg/kg) 45 min prior and immediately after SAH as evaluated by Western blot. Application of KN93 at 1 h and every 12 h post-SAH significantly reduced vascular CaMKII immunoreactivity at 72 h. In addition, contractile responses of cerebral arteries to endothelin-1 (ET-1) and 5-hydroxycarboxamide (5-CT) were increased at this time-point. KN93 treatment significantly attenuated the contraction induced by ET-1 and 5-CT. Importantly, treatment with the CaMKII inhibitor prevented SAH-induced deficits in neurological function, as evaluated by the rotating pole test, and similar sensorimotor scores were seen in sham-operated animals. The present study has shown that SAH is associated with increased contractile responses to ET-1 and 5-CT in cerebral arteries and enhanced early activation of CaMKII. Treatment with the CaMKII inhibitor KN93 attenuated the contractile responses and prevented impaired sensorimotor function after SAH.


Age of onset and type of diabetes.
The age of onset and the clinical type of diabetes mellitus were evaluated on the basis of a cross-sectional study of medical records of 14 municipal health centers in East Finland. Altogether 281 patients were classified as having insulin-dependent (IDDM) and 2713 as having non-insulin-dependent (NIDDM) diabetes. Nearly all patients diagnosed before the age of 19 had IDDM, but a large proportion (37%) of all diagnoses of IDDM were made after that age. Six percent of all diabetic subjects in the age group 15-19 yr were classified as NIDDM and the proportion increased rapidly in older age groups. Half of the patients with NIDDM were diagnosed over the age of 64.


Developmental status at age 12 months according to birth weight and family income: a comparison of two Brazilian birth cohorts.
Two cohorts of children born in the city of Pelotas, Southern Brazil, in 1993 and 2004, were compared in terms of neuro-psychomotor development at the age of 12 months. Children were evaluated using the Denver II screening test. Analyses were performed using the Poisson regression technique. The prevalence of suspected developmental delay fell from 37.1% in 1993 to 21.4% in 2004 and was inversely proportional to family income and birth weight. Among children born weighing under 2,000 g, there was a fourfold reduction in the prevalence of developmental delay between 1993 and 2004. With regard to family income, the poorest group showed the greatest reduction between the two cohorts--a 30% reduction in risk. Our results confirm the influence of income and birth weight on child development. The decrease in the prevalence of developmental delay in the last decade reflects, among other factors, improvements in neonatal care, increased coverage of developmental monitoring in the first year of life, and longer breastfeeding duration. Despite this reduction, the prevalence of developmental delay is still high, reinforcing the need for early diagnosis and intervention.


Comparative efficacy of family and group treatment for adolescent substance abuse.
Due to the continuing prevalence of adolescent substance abuse, promising treatment models need to be developed and evaluated. Thus, the purpose of this study was to evaluate the efficacy of two promising models, Strengths Oriented Family Therapy (SOFT) and The Seven Challenges(R) (7C). Adolescents who qualified for outpatient treatment and agreed to participate in our study were randomly assigned to one of the two treatments and assessed at 3 and 6-months following baseline. Using a two-part, random-effects model, we examined the odds of achieving abstinence or full symptom remission between treatments and over time. For those not achieving full abstinence or full problem remission, we investigated whether frequency of use or symptom severity were reduced at follow-up. Participants in both SOFT and 7C demonstrated significant reductions in substance use and related problems, but treatments did not differ at 3 and 6 months following baseline. Overall, treatment services were delivered as planned. Both SOFT and 7C were efficacious with adolescents who abuse substances, as participants in both conditions were significantly more likely to be in symptom remission or abstinent at follow-up interviews versus at baseline. Replication studies are needed that address this study's limitations.


Antiatherosclerotic effects of dietary supplementations of garlic and turmeric: Restoration of endothelial function in rats.
Protections of endothelial integrity by elimination of certain risk have proven to be effective in maintaining hemostasis and in slowing the progress of the cardiovascular disease. Indigenous drugs are the natural source of protection against these disorders, which can be used more effectively by the knowledge of their active ingredients as well as by their mechanism of action. Most prominent among these drugs are garlic, [Alium sativum L., Family: Liliaceae, Bulbs] and turmeric [Curcuma longa L., Family: Zingiberaceae, Rhizomes]; commonly used Indian traditional spices. In the present study, we examined the atheroscleroprotective potential of diet supplementation of garlic and turmeric by measuring serum lipid profile, changes in cardiovascular parameters i.e. arterial blood pressure, electrocardiogram and heart rate. We further tried to elucidate the mechanism of restoration of endothelial function and the role of endothelium-derived factors mainly, nitric oxide (NO) and cycloxygenase derived contracting factors. A notable restoration of arterial blood pressure was seen in animals on garlic and turmeric supplemented diet. Animals on supplemented diet showed a significantly enhanced vasorelaxant response to adenosine, acetylcholine, isoproterenol and contractile effect of 5-hyderoxytryptamine was significantly attenuated. Inhibition of these responses by L-NMMA was smaller in tissues from herbal treated animals. Incubation of tissues with L-arginine (10(-5) M) resulted in a significant reversal of L-NMMA induced inhibition of endothelium-mediated relaxation, which appeared to be pronounced in rings from animals supplemented with herbs as compared to hypercholesterolemic animals. Addition of indomethacin (10(-5) M) augmented the relaxation in all the groups of animals. The present study demonstrated that garlic and turmeric are potent vasorelaxants as well as reduce the atherogenic properties of cholesterol. Whether combination of these vasodilators in cardiovascular disorders with increased peripheral vascular resistance remains to be determined.


Adaptative biaryl phosphite-oxazole and phosphite-thiazole ligands for asymmetric Ir-catalyzed hydrogenation of alkenes.
A library of readily available phosphite-oxazole/thiazole ligands (L1 a-g-L7 a-g) was applied in the Ir-catalyzed asymmetric hydrogenation of several largely unfunctionalized E- and Z-trisubstituted and 1,1-disubstituted terminal alkenes. The ability of the catalysts to transfer chiral information to the product could be tuned by choosing suitable ligand components (bridge length, the substituents in the heterocyclic ring and the alkyl backbone chain, the configuration of the ligand backbone, and the substituents/configurations in the biaryl phosphite moiety), so that enantioselectivities could be maximized for each substrate as required. Enantioselectivities were therefore excellent (enantiomeric excess (ee) values up to >99 %) for a wide range of E- and Z-trisubstituted and 1,1-disubstituted terminal alkenes. The biaryl phosphite moiety was a very advantageous ligand component in terms of substrate versatility.


Granulocyte colony--stimulating factor for chemotherapy-induced neutropenia in patients with small cell lung cancer : the 40% rule revisited.
Recombinant granulocyte colony-stimulating factor (G-CSF) [filgrastim and lenograstim] and pegylated G-CSF (pegfilgrastim) have been shown to reduce the severity and duration of chemotherapy-associated febrile neutropenia (FN) when administered prophylactically to cancer patients receiving chemotherapeutic regimens. The American Society of Clinical Oncology (ASCO) evidence-based clinical guidelines published in 1994, 1996 and 1997 recommended primary prophylaxis with G-CSF for cancer patients. The 2000 ASCO update, with the same recommendation, highlights the importance of economic considerations in decision making for CSFs. This paper reviews the available cost-effectiveness evidence on the use of G-CSF as primary prophylaxis against FN in patients with small cell lung cancer (SCLC).Cost-effectiveness ratios from a healthcare payer perspective supported the use of filgrastim as primary prophylaxis for people with SCLC, on the basis of both clinical and economic benefits, treated with chemotherapeutic regimens that have an FN rate in the range of 40-60%. However, when indirect and patient out-of-pocket costs attributable to severe FN are included, available evidence suggests that the risk threshold may be reduced by more than half. Given that FN rates associated with chemotherapeutic regimens for SCLC are generally <40%, then few circumstances would warrant the use of G-CSFs (filgrastim and lenograstim) under the current rule. However, inclusion of indirect costs would lower the cost-effectiveness threshold. Future cost-effectiveness studies of medications such as pegfilgrastim should attempt to capture the societal perspective by incorporating productivity-related costs and using base-case rates of FN reported in the literature.


Cognitive Remediation in Bipolar (CRiB2): study protocol for a randomised controlled trial assessing efficacy and mechanisms of cognitive remediation therapy compared to treatment as usual.
A substantial proportion of people with bipolar disorder (BD) experience persistent cognitive difficulties associated with impairments in psychosocial functioning and a poorer disorder course. Emerging evidence suggests that cognitive remediation (CR), a psychological intervention with established efficacy in people with schizophrenia, can also benefit people with BD. Following a proof-of-concept trial showing that CR is feasible and potentially beneficial for people with BD, we are conducting an adequately powered trial in euthymic people with BD to 1) determine whether an individual, therapist-supported, computerised CR can reduce cognitive difficulties and improve functional outcomes; and 2) explore how CR exerts its effects. CRiB2 is a two-arm, assessor-blind, multi-site, randomised controlled trial (RCT) comparing CR to treatment-as-usual (TAU). Participants are people with a diagnosis of BD, aged between 18 and 65, with no neurological or current substance use disorder, and currently euthymic. 250 participants will be recruited through primary, secondary, tertiary care, and the community. Participants will be block-randomised (1:1 ratio, stratified by site) to continue with their usual care (TAU) or receive a 12-week course of therapy and usual care (CR + TAU). The intervention comprises one-on-one CR sessions with a therapist supplemented with independent cognitive training for 30-40 h in total. Outcomes will be assessed at 13- and 25-weeks post-randomisation. Efficacy will be examined by intention-to-treat analyses estimating between-group differences in primary (i.e., psychosocial functioning at week 25 measured with the Functional Assessment Short Test) and secondary outcomes (i.e., measures of cognition, mood, patient-defined goals, and quality of life). Global cognition, metacognitive skills, affect fluctuation, and salivary cortisol levels will be evaluated as putative mechanisms of CR through mediation models. This study will provide a robust evaluation of efficacy of CR in people with BD and examine the putative mechanisms by which this therapy works. The findings will contribute to determining the clinical utility of CR and potential mechanisms of action. Cognitive Remediation in Bipolar 2 (CRiB2): ISRCTN registry: https://www.isrctn.com/ISRCTN10362331 . Registered 04 May 2022. Overall trial status: Ongoing; Recruitment status: Recruiting.


Differential expression of transforming growth factor-beta receptors in a rabbit zone II flexor tendon wound healing model.
Flexor tendon repair in zone II is complicated by adhesions that impair normal postoperative gliding. Transforming growth factor-beta (TGF-beta) is a family of growth factors that has been implicated in scar formation. The TGF-beta family of proteins binds to three distinct classes of membrane receptors, termed RI, RII, and RIII. In this study, we analyzed the temporal and spatial distribution of TGF-beta receptor isoforms (RI, RII, and RIII) in a rabbit zone II flexor tendon wound healing model.Twenty-eight adult New Zealand White rabbit forepaws underwent isolation of the middle digit flexor digitorum profundus tendon in zone II. The tendons underwent transection in zone II and immediate repair. The tendons were harvested at increasing time points: 1, 3, 7, 14, 28, and 56 days postoperatively (n = 4 at each time point). The control flexor tendons were harvested without transection and repair (n = 4). Immunohistochemical analysis was used to detect the expression patterns for TGF-beta receptors RI, RII, and RIII. Immunohistochemical staining of the transected and repaired tendons demonstrated up-regulation of TGF-beta RI, RII, and RIII protein levels. TGF-beta receptor production in the experimental group (transection and repair) was concentrated in the epitenon and along the repair site. Furthermore, the TGF-beta receptor expression levels peaked at day 14 and decreased by day 56 postoperatively. In contrast, minimal receptor expression was observed in the untransected and unrepaired control tendons. These data provide evidence that (1) TGF-beta receptors are up-regulated after injury and repair; (2) peak levels of TGF-beta receptor expression occurred at day 14 and decreased by day 56 after wounding and repair; and (3) both the tendon sheath and epitenon have the highest receptor expression, and both may play critical roles in flexor tendon wound healing. Understanding the up-regulation of TGF-beta isoforms and the up-regulation of their corresponding receptors during flexor tendon wound healing provides new targets for biomolecular modulation of postoperative scar formation.


The sak1+ gene of Schizosaccharomyces pombe encodes an RFX family DNA-binding protein that positively regulates cyclic AMP-dependent protein kinase-mediated exit from the mitotic cell cycle.
In Schizosaccharomyces pombe, meiosis is initiated by conditions of nutrient deprivation. Mutations in genes encoding elements of the cyclic AMP-dependent protein kinase (cAPK) pathway interfere with meiosis. Loss-of-function alleles of genes that stimulate the activity of cAPK allow cells to bypass the normal requirement of starvation for conjugation and meiosis. Alternatively, loss-of-function alleles of genes that inhibit cAPK lead to the inability to undergo sexual differentiation. The cgs1+ gene encodes the regulatory subunit of cAPK, and the cgs2+ gene encodes a cyclic AMP phosphodiesterase. Thus, both genes encode proteins which negatively regulate the activity of cAPK. Loss of either cgs1 or cgs2 prevents haploid cells from conjugating and diploid cells from undergoing meiosis. In addition to these defects, cells are unable to enter stationary phase. We describe a novel gene, sak1+, which when present on a plasmid overcomes the aberrant phenotypes associated with unregulated cAPK activity. Genetic analysis of sak1+ (suppressor of A-kinase) reveals that it functions downstream of cyclic AMP-dependent protein kinase to allow cells to exist the mitotic cycle and enter either stationary phase or the pathway leading to sexual differentiation. The sak1+ gene is essential for cell viability, and a null allele causes multiple defects in cell morphology and nuclear division. Thus, sak1+ is an important regulatory element in the life cycle of S. pombe. Sequence analysis shows that the predicted product of the sak1+ gene is an 87-kDa protein which shares homology to the RFX family of DNA-binding proteins identified in humans and mice. One member of this family, RFX1, is a transcription factor for a variety of viral and cellular genes.


[Continuing medical professional development in Uruguay as the country faces the 21st century].
This paper outlines the evolution of the concept of continuing medical professional development in Uruguay, as that professional development has progressed from independent and varied efforts that were not coordinated or evaluated to a plan that is aimed at improving the quality of educational activities and programs and making them accessible to all the physicians in the country. In contrast to countries where scientific societies and medical associations preside over continuing professional development, in Uruguay the Graduate School of the School of Medicine of the University of the Republic has taken on managing and developing this process. The key objective of this process is for the Graduate School to work in conjunction with other leading players in professional development to create a national system of continuing medical professional development. This system should be interinstitutional and include both public and private entities. By having the key public institutions (the Graduate School and the Ministry of Public Health) working with private entities (medical associations, scientific societies, and health care institutions), there should be an adequate balance of interests. The national professional development system should work in the most decentralized manner possible and should be based on a network of units distributed around the entire country, but with centralized coordination. The system's interinstitutional character should ensure high technical and ethical standards as well as a balance among the governmental, commercial, professional, and university components. This system should serve as the basis for extending this activity to all health personnel in Uruguay. While the national system is still being finalized, the Graduate School has implemented a process of accrediting institutions that are involved in continuing professional medical development. The aim of this accreditation process is to improve the educational offerings for physicians and to generate practical experience that will serve the future system.


On the identifiability of metabolic network models.
A major problem for the identification of metabolic network models is parameter identifiability, that is, the possibility to unambiguously infer the parameter values from the data. Identifiability problems may be due to the structure of the model, in particular implicit dependencies between the parameters, or to limitations in the quantity and quality of the available data. We address the detection and resolution of identifiability problems for a class of pseudo-linear models of metabolism, so-called linlog models. Linlog models have the advantage that parameter estimation reduces to linear or orthogonal regression, which facilitates the analysis of identifiability. We develop precise definitions of structural and practical identifiability, and clarify the fundamental relations between these concepts. In addition, we use singular value decomposition to detect identifiability problems and reduce the model to an identifiable approximation by a principal component analysis approach. The criterion is adapted to real data, which are frequently scarce, incomplete, and noisy. The test of the criterion on a model with simulated data shows that it is capable of correctly identifying the principal components of the data vector. The application to a state-of-the-art dataset on central carbon metabolism in Escherichia coli yields the surprising result that only 4 out of 31 reactions, and 37 out of 100 parameters, are identifiable. This underlines the practical importance of identifiability analysis and model reduction in the modeling of large-scale metabolic networks. Although our approach has been developed in the context of linlog models, it carries over to other pseudo-linear models, such as generalized mass-action (power-law) models. Moreover, it provides useful hints for the identifiability analysis of more general classes of nonlinear models of metabolism.


Correlates of subjective well-being in schizophrenic patients treated with atypical antipsychotics.
Objective A growing body of research indicates that a low subjective well-being (SW) may be predictive of non-adherence and less favourable outcome. This study examined baseline variables and variables in the course of treatment hypothesised to be associated with later SW. Methods Sixty-three inpatients with schizophreniform disorder or schizophrenia were randomly assigned to treatment with various atypical antipsychotics after a wash-out phase of 2 days. Subjects were evaluated with a protocol that examined psychopathology (Positive and Negative Symptom Scale, PANSS), side effects (Scandinavian Society of Pharmacology, UKU), and subjective well-being (Subjective Well-being under Neuroleptic treatment, SWN) at baseline and endpoint (mean duration of treatment 39.9 days). Two-thirds of subjects were multiple episode schizophrenic inpatients pre-treated with antipsychotics. Results Multiple regression analyses revealed that the PANSS negative score, neurological side effects, and SWN at baseline, as well as change of the PANSS positive score between baseline and endpoint, were associated independently with SW at endpoint (R(2)=0.55 after exclusion of two subjects). Conclusions Patients with low SW, severe negative symptoms, and neurological side effects, all at baseline, as well as those without improvement or deterioration of positive symptoms are at risk of low SW later in treatment and, most likely, of non-adherence.


Psychometric characteristics of the mental health crisis assessment scale in youth with autism spectrum disorder.
Youth with autism spectrum disorder (ASD) exhibit high rates of psychopathology. These symptoms can pose a risk of injury to self or others when the child is in crisis. Despite this danger, there are no instruments available to identify those with ASD who are at risk or actively in crisis. This study examined the psychometric properties of the Mental Health Crisis Assessment Scale (MCAS), a 28 item parent report measure. The MCAS was administered to the parents of 606 children and young adults (aged 3-25 years, M age = 13 years, SD = 5 years) enrolled in the Interactive Autism Network, an online registry of families raising a child with ASD. The MCAS asks parents to rate the severity of various emotional and behavioral symptoms exhibited by their child. The parent then selects the behavior they perceive as the most dangerous behavior and rates the acuity of as well as their efficacy in managing this behavior. The MCAS was tested for internal consistency, construct validity, criterion validity, and convergent validity. The MCAS demonstrated strong internal consistency (Total Scale Cronbach's α = .88). The exploratory and confirmatory factor analyses suggested that a two factor (acuity and behavioral efficacy) model fit the data well, providing evidence of construct validity. Criterion validity, which was assessed by comparing the MCAS to clinician determination of crisis, indicated high levels of agreement (ROC = .85). Strong positive relationships emerged between the MCAS and measures of family distress (r = .56), parental stress, and frustration (r = .48), and use of emergency psychiatric services (OR = 24.2, 95% CI: 8.6-68.2), indicating convergent validity of the measure (all p < .05). Results of the psychometric analyses suggest the MCAS appears to be a promising tool that can measure mental health crises in youth with ASD.


Gene-expression variation within and among human populations.
Understanding patterns of gene-expression variation within and among human populations will provide important insights into the molecular basis of phenotypic diversity and the interpretation of patterns of expression variation in disease. However, little is known about how gene-expression variation is apportioned within and among human populations. Here, we characterize patterns of natural gene-expression variation in 16 individuals of European and African ancestry. We find extensive variation in gene-expression levels and estimate that approximately 83% of genes are differentially expressed among individuals and that approximately 17% of genes are differentially expressed among populations. By decomposing total gene-expression variation into within- versus among-population components, we find that most expression variation is due to variation among individuals rather than among populations, which parallels observations of extant patterns of human genetic variation. Finally, we performed allele-specific quantitative polymerase chain reaction to demonstrate that cis-regulatory variation in the lymphocyte adaptor protein (SH2B adapter protein 3) contributes to differential expression between European and African samples. These results provide the first insight into how human population structure manifests itself in gene-expression levels and will help guide the search for regulatory quantitative trait loci.


Successful implantation of bioengineered, intrinsically innervated, human internal anal sphincter.
To restore fecal continence, the weakened pressure of the internal anal sphincter (IAS) must be increased. We bioengineered intrinsically innervated human IAS to emulate sphincteric physiology in vitro. We cocultured human IAS circular smooth muscle with immortomouse fetal enteric neurons. We investigated the ability of bioengineered innervated human IAS, implanted in RAG1-/- mice, to undergo neovascularization and preserve the physiology of the constituent myogenic and neuronal components. The implanted IAS was neovascularized in vivo; numerous blood vessels were observed with no signs of inflammation or infection. Real-time force acquisition from implanted and preimplant IAS showed distinct characteristics of IAS physiology. Features included the development of spontaneous myogenic basal tone; relaxation of 100% of basal tone in response to inhibitory neurotransmitter vasoactive intestinal peptide (VIP) and direct electrical field stimulation of the intrinsic innervation; inhibition of nitrergic and VIPergic electrical field-induced relaxation (by antagonizing nitric oxide synthesis or receptor interaction); contraction in response to cholinergic stimulation with acetylcholine; and intact electromechanical coupling (evidenced by direct response to potassium chloride). Implanted, intrinsically innervated bioengineered human IAS tissue preserved the integrity and physiology of myogenic and neuronal components. Intrinsically innervated human IAS bioengineered tissue can be successfully implanted in mice. This approach might be used to treat patients with fecal incontinence.


5-Aminolevulinic acid induced protoporphyrin IX as a fluorescence marker for quantitative image analysis of high-grade dysplasia in Barrett's esophagus cellular models.
Early detection and treatment of high-grade dysplasia (HGD) in Barrett's esophagus may reduce the risk of developing esophageal adenocarcinoma. Confocal endomicroscopy (CLE) has shown advantages over routine white-light endoscopic surveillance with biopsy for histological examination; however, CLE is compromised by insufficient contrast and by intra- and interobserver variation. An FDA-approved PDT photosensitizer was used here to reveal morphological and textural features similar to those found in histological analysis. Support vector machines were trained using the aforementioned features to obtain an automatic and robust detection of HGD. Our results showed 95% sensitivity and 87% specificity using the optimal feature combination and demonstrated the potential for extension to a three-dimensional cell model.


Ethics of genetic and biomarker test disclosures in neurodegenerative disease prevention trials.
Prevention trials for neurodegenerative diseases use genetic or other risk marker tests to select participants but there is concern that this could involve coercive disclosure of unwanted information. This has led some trials to use blinded enrollment (participants are tested but not told of their risk marker status). We examined the ethics of blinded vs transparent enrollment using well-established criteria for assessing the ethics of clinical research. Normative analysis applying 4 key ethical criteria-favorable risk-benefit ratio, informed consent, fair subject selection, and scientific validity-to blinded vs transparent enrollment, using current evidence and state of Alzheimer disease (AD) and other prevention trials. Current evidence on the psychosocial impact of risk marker disclosure and considerations of scientific benefit do not support an obligation to use blinded enrollment in prevention trials. Nor does transparent enrollment coerce or involve undue influence of potential participants. Transparent enrollment does not unfairly exploit vulnerable participants or limit generalizability of scientific findings of prevention trials. However, if the preferences of a community of potential participants would affect the rigor or feasibility of a prevention trial using transparent enrollment, then investigators are required by considerations of scientific validity to use blinded enrollment. Considerations of risks and benefits, informed consent, and fair subject selection do not require the use of blinded enrollment for AD prevention trials. Blinded enrollment in AD prevention trials may sometimes be necessary because of the need for scientific validity, not because it prevents coercion or undue influence.


Low-grade astrocytoma with neuraxis dissemination at diagnosis.
Little is known about low-grade astrocytoma with neuraxis dissemination at diagnosis. A review of medical records identified this phenomenon in eight of 150 pediatric patients evaluated between 1985 and 1994 for histologically confirmed low-grade astrocytoma. These patients (five male and three female) ranged in age from 5 months to 20 years (median 8 years). Symptoms of neuraxis disease were minimal or absent. Primary tumor sites were the hypothalamus in four cases, brainstem/spinal cord in three, and temporal lobe in one. Patterns of dissemination (evaluated by computerized tomography and/or magnetic resonance imaging techniques) appeared to be related to the primary site: hypothalamic tumors metastasized along the ventricular cerebrospinal fluid pathways, and tumors in other locations disseminated along subarachnoid pathways. Following initial treatment with chemotherapy (in three), partial resection (in one), radiation therapy (in three), and chemotherapy plus irradiation (in one), four patients required salvage therapy for progressive or recurrent disease. Seven of the eight patients are alive with stable or progressive disease 6 to 105 months postdiagnosis (median 15 months). Low-grade astrocytoma with initial neuraxis dissemination is responsive to chemotherapy and radiation, a proportion showing periods of stable disease. The optimum therapy or combination of therapies remains unclear.


Acute care hospitalization near the end of life for cancer patients who die in hospital in Canada.
Acute care hospitals have a role in managing the health care needs of people affected by cancer when they are at the end of life. However, there is a need to provide end-of-life care in other settings, including at home or in hospice, when such settings are more appropriate. Using data from 9 provinces, we examined indicators that describe the current landscape of acute care hospital use at the end of life for patients who died of cancer in hospital in Canada. Interprovincial variation was observed in acute care hospital deaths, length of stay in hospital, readmission to hospital, and intensive care unit use at the end of life. High rates of acute care hospital use near the end of life might suggest that community and home-based end-of-life care might not be suiting patient needs.


Heat shock protein 90 inhibitors augment endogenous wild-type p53 expression but down-regulate the adenovirally-induced expression by inhibiting a proteasome activity.
Heat shock protein 90 (HSP90) inhibitors suppressed MDM4 functions which mediated p53 ubiquitination, and blocked a chaperon function which influenced expression of the client proteins. We examined cytotoxic effects of the inhibitors, 17-allylamino-17-demetheoxygeldanamycin (17-AAG) and 17-dimethylaminoethylamino-17-demethoxy-geldanamycin (17-DMAG), on mesothelioma and investigated combinatory effects of the inhibitors and adenoviruses expressing the wild-type p53 gene (Ad-p53). A majority of mesothelioma lacks p14 and p16 expression, which leads to defective p53 pathway despite bearing the wild-type p53 genotype. The HSP90 inhibitors up-regulated endogenous wild-type p53 expression and induced cell death. Furthermore, the inhibitors increased the endogenous p53 levels that were induced by cisplatin. Nevertheless, the HSP90 inhibitors suppressed Ad-p53-induced exogenous p53 expression primarily at a posttranscriptional level and inhibited the Ad-p53-mediated cell death. HSP90 inhibitors suppressed ubiquitination processes which were involved in p53 degradation, but a proteasome inhibitor, MG-132, prevented the HSP90 inhibitors-induced p53 down-regulation. In contrast, an inhibitor for HSP70 with a chaperon function, pifithrin-μ, did not produce the p53 down-regulation. The HSP90 inhibitors did not suppress expression of Ad receptor molecules but rather increased expression of green fluorescence protein transduced by the same Ad vector. These data collectively indicated that an HSP90 inhibitor possessed a divalent action on p53 expression, as an activator for endogenous wild-type p53 through inhibited ubiquitination and a negative regulator of exogenously over-expressed p53 through the proteasome pathway.


Utilizing Experimental Mouse Model to Identify Effectors of Hepatocellular Carcinoma Induced by HBx Antigen.
Hepatocellular carcinoma (HCC) is among the ten most commonly diagnosed cancers and the fourth leading cause of cancer-related death. Patients with hepatitis B virus (HBV) infection are prone to developing chronic liver diseases (i.e., fibrosis and cirrhosis), and the HBV X antigen plays an important role in the development of HCC. The difficulty in detecting HCC at the early stages is one of the main reasons that the death rate approximates the incidence rate. The regulators controlling the downstream liver protein expression from HBV infection are unclear. Mass spectrometric techniques and customized programs were used to identify differentially expressed proteins which may be involved in the development of liver fibrosis and HCC progression in hepatitis B virus X protein transgenic mice (HBx mice). FSTL1, CTSB, and TGF-β enhanced the signaling pathway proteins during the pathogenesis of HBx. Missing proteins can be essential in cell growth, differentiation, apoptosis, migration, metastasis or angiogenesis. We found that LHX2, BMP-5 and GDF11 had complex interactions with other missing proteins and BMP-5 had both tumor suppressing and tumorigenic roles. BMP-5 may be involved in fibrosis and tumorigenic processes in the liver. These results provide us an understanding of the mechanism of HBx-induced disorders, and may serve as molecular targets for liver treatment.


Translating the research diagnostic criteria for temporomandibular disorders into German: evaluation of content and process.
To develop a German-language version of the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) through a formal translation/back-translation process, to summarize available data about their psychometric properties, and to provide new data about psychometric testing of components of the RDC/TMD. To cross-culturally adapt the instrument, the RDC/TMD were translated using a forward-backward method, except for measures of somatization and depression, because German-specific instruments of these already existed. The psychometric properties of the RDC/TMD were examined, and the literature on this topic was reviewed. The available literature about reliability of clinical examination methods (4 studies) showed at least acceptable results, with a median intraclass correlation coefficient (ICC) of 0.60. Reliability of RDC/TMD components Jaw Disability List (JDL) and Graded Chronic Pain Scale (GCPS) was sufficient (ICC for retest reliability [n = 27] was 0.76 for JDL and 0.92 for GCPS; Cronbach's alpha for internal consistency [n = 378] was 0.72 and 0.88, respectively). A priori hypothesized associations between GCPS or JDL summary scores and self-report of general health, oral health, oral health-related quality of life, or dysfunctional pain, which were measured by means of the Multidimensional Pain Inventory, were confirmed in a convenience sample of clinical TMD patients (n = 378). These correlations were interpreted as support for the validity of the GCPS and JDL. The original RDC/TMD include measures for somatization and depression (SCL-90-R); however, equivalent German instruments to assess these constructs ("Beschwerdenliste," "Allgemeine Depressionsskala") have well-established validity and reliability in the German cultural environment. The psychometric properties and international comparability of the German version of the RDC/TMD (RDC/TMD-G) make this instrument suitable for the assessment of TMD in Germany.


Proteomics of Alzheimer's disease: understanding mechanisms and seeking biomarkers.
Alzheimer's disease is the scourge of the modern, aging world: a costly, damaging disease that robs the elderly of their ability to function as well as their memories. Three decades of progress have resulted in a deep understanding of the pathological processes and a range of targets for therapy, many of which have advanced to late-stage clinical trials. Proteomics has contributed greatly to these advances and will continue to have a growing role in determining the nature of the pathological lesions in the brain. In addition, proteomics (both gel based and gel free, mass spectrometry based), is likely to play an increasing role in identifying biomarkers that may assist in early diagnosis and in monitoring progression and, most importantly, response to therapy.


COVID-19 and the state: Exploring a puzzling relationship in the early stages of the pandemic.
During the first year of the Covid-19 pandemic, it was wealthier countries with stronger institutions that suffered the highest numbers of cases and fatalities. Many weaker countries were instead praised for more effective pandemic response. What explains this seeming puzzle? We re-consider these relationships in the cross-country data, drawing on measures of the state, Covid's health impact and pandemic response. In brief, our analysis suggests that, when appropriate additional factors are taken into account, the expected relationship between state effectiveness and pandemic health outcomes in fact is clear. We also offer insight into how different dimensions of the state influence policy and outcomes and how particular countries compare with others.


Estrogen induces neurotensin/neuromedin N messenger ribonucleic acid in a preoptic nucleus essential for the preovulatory surge of luteinizing hormone in the rat.
Ovarian steroids act on unidentified neurons to trigger preovulatory secretion of GnRH. In the rat, important steroid target cells reside in the anterior medial preoptic nucleus (AMPN), a sexually dimorphic structure essential for stimulatory effects of ovarian steroids on LH secretion. The AMPN contains neurotensin (NT)-immunoreactive neurons, and immunoneutralization of NT in the preoptic region markedly attenuates steroid-induced LH surges. Using probes derived from the rat gene that encodes NT and neuromedin N (NT/N), we investigated the ability of estrogen to influence NT/N mRNA levels in the AMPN. Ovariectomized rats were treated for 14 days with sham capsules or capsules that produce supraphysiological serum levels of 17 beta-estradiol (250 +/- 20 pg/ml). As determined by in situ hybridization, estradiol markedly altered the distribution of NT/N mRNA in the medial preoptic region, causing a striking increase in NT/N mRNA abundance specifically in the AMPN and adjacent medial preoptic nucleus (MPN). In contrast, estradiol caused no obvious changes in labeling in the lateral septum, diagonal band of Broca, bed nucleus of the stria terminalis, and lateral preoptic area. The distribution of NT/N mRNA in the AMPN of normal male rats closely resembled that in ovariectomized rats, where labeled cells were rarely observed. Microdissection and S1 nuclease protection analysis were used to quantitate the effect of estradiol on NT/N mRNA levels. Supraphysiological estradiol treatment for 14 days caused a 3.4-fold increase (P less than 0.0002) in NT/N mRNA levels in the combined AMPN/MPN, whereas levels in the central amygdaloid nucleus remained constant, providing further evidence of regional specificity. Forty-eight hours of estradiol treatment, at concentrations (60 +/- 1 pg/ml) similar to those observed on the morning of proestrus, caused a 1.8-fold increase (P less than 0.001) in NT/N mRNA levels in the AMPN/MPN, indicating that the time course of NT/N mRNA induction by estrogen is compatible with events of the normal estrous cycle. Together with previous findings, our results strongly suggest that NT neurons mediate, directly or indirectly, stimulatory effects of ovarian steroids on GnRH secretion.


Investigating community-based health and health promotion for homeless people: a mixed methods review.
Homeless people are susceptible to a range of health problems, yet in terms of health promotion, tend to be a hard-to-reach, marginalized group. Robust evidence regarding the ability to engage with this population via effective health promotion programmes is essential if policy and practice are to be informed to improve the health of homeless people. A structured review was conducted with the aim of examining what is known about community-based health promotion for homeless people. Six databases were searched and 8435 records screened. Thirteen studies met the inclusion criteria. A mixed-methods 'combined separate synthesis' approach was used to accommodate both quantitative and qualitative evidence within one review. Three themes emerged: (i) incorporating homelessness, (ii) health improving and (iii) health engaging. The review has implications for health promotion design, with evidence suggesting that as part of a tailored approach, homeless people must be actively involved in intervention development, ensuring that appropriate, acceptable and potentially effective individual elements are incorporated into community-based interventions.


A quantitative structure-property relationship for predicting drug solubility in PEG 400/water cosolvent systems.
A quantitative structure-property relationship (QSPR) was developed to predict drug solubility in binary mixtures of polyethylene glycol (PEG) 400 and water. The ability of the QSPR model to predict solubility was assessed and compared to the classic log-linear cosolvency model. The solubility of 122 drugs, ranging in log P from -2.4 to 7.5, was determined in 0%, 25%, 50%, and 75% PEG (v/v in water) by the shake-flask method. Solubility data from 84 drugs were fit by linear regression using the following molecular descriptors: molecular weight, volume, radius of gyration, density, number of rotatable bonds, hydrogen-bond donors, and hydrogen-bond acceptors. The multiple linear regression model was optimized by a genetic algorithm guided selection method. The remaining 38 compounds were used to test the predictability of the model. QSPR-based models developed at each volume fraction with the training set compounds showed a reasonable correlation coefficient (r) of approximately 0.9 and a root mean square (rms) error of <0.5 log unit. The model predicted solubility values of approximately 78% of the testing set compounds within 1 log unit. The log-linear model was as effective as the QSPR-based model in predicting the testing set solubilities; however, many drugs, as expected, showed significant deviation from log-linearity. The QSPR model requires only the chemical structure of the drug and has utility for guiding vehicle identification for early preclinical in vivo studies, especially when compound availability is limited and experimental data such as aqueous solubility and melting point are unknown. When experimental data are available, the log-linear model was verified to be a useful predictive tool.


Histamine and migraine revisited: mechanisms and possible drug targets.
To review the existing literature on histamine and migraine with a focus on the molecule, its receptors, its use in inducing migraine, and antihistamines in the treatment of migraine. Histamine has been known to cause a vascular type headache for almost a hundred years. Research has focused on antihistamines as a possible treatment and histamine as a migraine provoking agent but there has been little interest in this field for the last 25 years. In recent years two additional histamine (H3 and H4) receptors have been discovered and a series of non-sedating antihistamines have been developed. It is therefore timely to review the field again. For this review the PubMed/MEDLINE database was searched for eligible studies. We searched carefully for all articles on histamine, antihistamines and histamine receptors in relation to migraine and the nervous system. The following search terms were used: histamine, migraine disorders, migraine, headache, antihistamines, histamine antagonists, clinical trials, induced headache, histamine H3 receptor, histamine H4 receptor and pharmacology. Four hundred thirty-six titles were read, 135 abstracts were read, 112 articles were read in full and 53 articles were used in this review. Review process resulted in 12 articles added to a total of 65. Early studies of H1 and H2 antihistamines lack scientific strength and show conflicting results. Most of the antihistaminic drugs used in these trials bind also to other receptors which makes it difficult to conclude on the antihistaminic effect. Histamine is an efficient inducer of migraine attacks in migraine patients by an H1 mechanism most likely extracerebrally. These findings merit further investigation of antihistamines in clinical drug trials. The H3 and H4 receptors are found in primarily in CNS and immune tissues, respectively. H3 is likely to be involved in antinociception and has been linked with cognitive, neurodegenerative and sleep disorders. The only marketed H3 agent, pitolisant, is a brain penetrant H3 antagonist/inverse agonist which increases central histamine and causes headache. The experimental H3 agonist Nα-methylhistamine has shown promising results as a migraine preventative in studies of uncertain quality. With the current limited knowledge of the H4 receptor it is questionable whether or not the receptor is involved in migraine. There is insufficient support for first generation antihistamines (both H1 and H2) as preventive migraine medications and sedation and weight gain are unacceptable side effects. Non-sedating H1 antihistamines need to be appropriately tested. Central H3 receptors seem to have a role in migraine that merit further investigation. The histaminergic system may be a goal for novel migraine drugs.


Glaser-mediated synthesis and photophysical characterization of diphenylbutadiyne-linked porphyrin dyads.
The Pd-mediated Glaser coupling of a zinc monoethynyl porphyrin and a magnesium monoethynyl porphyrin affords a mixture of three 4,4'-diphenylbutadiyne-linked dyads comprised of two zinc porphyrins (Zn-pbp-Zn), two magnesium porphyrins (Mg-pbp-Mg), and one metalloporphyrin of each type (Zn-pbp-Mg). The latter is easily isolated due to the greater polarity of the magnesium versus the zinc chelate. Exposure of Zn-pbp-Mg to silica gel results in selective demetalation, affording Zn-pbp-Fb where Fb = free base porphyrin. This synthesis route employs the magnesium porphyrin as a latent form of the Fb porphyrin, thereby avoiding copper insertion during the Glaser reaction, and as a polar entity facilitating separation. The absorption spectrum of Zn-pbp-Mg or Zn-pbp-Fb is the sum of the spectra of the component parts, while in each case the fluorescence spectrum upon illumination of the Zn porphyrin is dominated by emission from the Mg or Fb porphyrin. Time-resolved absorption spectroscopy shows that the energy-transfer rate constants are (11 ps)(-1) and (37 ps)(-1) for Zn-pbp-Mg and Zn-pbp-Fb, respectively, corresponding to energy-transfer quantum yields of 0.995 and 0.983, respectively. The calculated Förster through-space rates are (1900 ps)(-1) and (1100 ps)(-1) for Zn-pbp-Mg and Zn-pbp-Fb, respectively. Accordingly, the through-bond process dominates for both dyads with a through-bond:through-space energy-transfer ratio of > or =97:1. Collectively, the studies show that the 4,4'-diphenylbutadiynyl linker supports fast and efficient energy transfer between Zn and Mg or Fb porphyrins.


Reanalysis of morphine consumption from two randomized controlled trials of gabapentin using longitudinal statistical methods.
Postoperative pain management in total joint replacement surgery remains ineffective in up to 50% of patients and has an overwhelming impact in terms of patient well-being and health care burden. We present here an empirical analysis of two randomized controlled trials assessing whether addition of gabapentin to a multimodal perioperative analgesia regimen can reduce morphine consumption or improve analgesia for patients following total joint arthroplasty (the MOBILE trials). Morphine consumption, measured for four time periods in patients undergoing total hip or total knee arthroplasty, was analyzed using a linear mixed-effects model to provide a longitudinal estimate of the treatment effect. Repeated-measures analysis of variance and generalized estimating equations were used in a sensitivity analysis to compare the robustness of the methods. There was no statistically significant difference in morphine consumption between the treatment group and a control group (mean effect size estimate 1.0, 95% confidence interval -4.7, 6.7, P=0.73). The results remained robust across different longitudinal methods. The results of the current reanalysis of morphine consumption align with those of the MOBILE trials. Gabapentin did not significantly reduce morphine consumption in patients undergoing major replacement surgeries. The results remain consistent across longitudinal methods. More work in the area of postoperative pain is required to provide adequate management for this patient population.


Metal-mediated affinity and orientation specificity in a computationally designed protein homodimer.
Computationally designing protein-protein interactions with high affinity and desired orientation is a challenging task. Incorporating metal-binding sites at the target interface may be one approach for increasing affinity and specifying the binding mode, thereby improving robustness of designed interactions for use as tools in basic research as well as in applications from biotechnology to medicine. Here we describe a Rosetta-based approach for the rational design of a protein monomer to form a zinc-mediated, symmetric homodimer. Our metal interface design, named MID1 (NESG target ID OR37), forms a tight dimer in the presence of zinc (MID1-zinc) with a dissociation constant <30 nM. Without zinc the dissociation constant is 4 μM. The crystal structure of MID1-zinc shows good overall agreement with the computational model, but only three out of four designed histidines coordinate zinc. However, a histidine-to-glutamate point mutation resulted in four-coordination of zinc, and the resulting metal binding site and dimer orientation closely matches the computational model (Cα rmsd = 1.4 Å).


Chromatin modification by PSC occurs at one PSC per nucleosome and does not require the acidic patch of histone H2A.
Chromatin architecture is regulated through both enzymatic and non-enzymatic activities. For example, the Polycomb Group (PcG) proteins maintain developmental gene silencing using an array of chromatin-based mechanisms. The essential Drosophila PcG protein, Posterior Sex Combs (PSC), compacts chromatin and inhibits chromatin remodeling and transcription through a non-enzymatic mechanism involving nucleosome bridging. Nucleosome bridging is achieved through a combination of nucleosome binding and self-interaction. Precisely how PSC interacts with chromatin to bridge nucleosomes is not known and is the subject of this work. We determine the stoichiometry of PSC-chromatin interactions in compact chromatin (in which nucleosomes are bridged) using Scanning Transmission Electron Microscopy (STEM). We find that full compaction occurs with one PSC per nucleosome. In addition to compacting chromatin, we show that PSC oligomerizes nucleosome arrays. PSC-mediated oligomerization of chromatin occurs at similar stoichiometry as compaction suggesting it may also involve nucleosome bridging. Interactions between the tail of histone H4 and the acidic patch of histone H2A are important for chromatin folding and oligomerization, and several chromatin proteins bind the histone H2A acidic patch. However, mutation of the acidic patch of histone H2A does not affect PSC's ability to inhibit chromatin remodeling or bridge nucleosomes. In fact, PSC does not require nucleosomes for bridging activity but can bridge naked DNA segments. PSC clusters nucleosomes on sparsely assembled templates, suggesting it interacts preferentially with nucleosomes over bare DNA. This may be due to the ability of PSC to bind free histones. Our data are consistent with a model in which each PSC binds a nucleosome and at least one other PSC to directly bridge nucleosomes and compact chromatin, but also suggest that naked DNA can be included in compacted structures. We discuss how our data highlight the diversity of mechanisms used to modify chromatin architecture.


Maternal discipline of young children: context, belief, and practice.
This report describes maternal discipline of infants and toddlers, focusing on how the context of the misbehavior affects maternal beliefs and practices. Mothers of 1- to 4-year-old children were interviewed using a 20-minute, structured questionnaire at two practice sites: (1) an inner-city teaching hospital pediatric clinic (n = 104) and (2) a private pediatrician's office in a nearby suburban neighborhood (n = 100). Measures were constructed to assess beliefs and practices about three kinds of discipline (teaching, spanking, and removing) and about two approaches to discipline (positive or negative). We found that mothers believe significantly (p < .001) more strongly in all three kinds of discipline for dangerous misbehavior compared with annoying ones. Belief in teaching, spanking, and negative approach increase with age; belief in removing decreases (p < .001); and belief in positive approach remains stable. Belief in spanking starts earlier than belief in teaching, and belief in negative tone increases with age. Mothers seem to have better formulated beliefs about spanking and negative and positive approach to discipline than about teaching or removing. Eighty-five percent believe time-out is at times appropriate for 2 year old, but only 67% had practiced it in the past week. Belief and practice of time-out varied by the age of the child and practice site. Explaining, distracting, and praising were practiced frequently, whereas bribing was less common. Less than half (44%) reported that they had ever discussed discipline with their child's physician, although 85% thought that their physician could give good advice. There is evidence that mothers change their minds about the appropriateness of time-out as their child gets older, but beliefs about other kinds of discipline remain relatively constant. We conclude that the type of misbehavior and the age of the child affect a mother's beliefs about discipline. These findings suggest a role for pediatricians in exploring and broadening parents' concepts of discipline.


Functional annotation of the transcriptome of Sorghum bicolor in response to osmotic stress and abscisic acid.
Higher plants exhibit remarkable phenotypic plasticity allowing them to adapt to an extensive range of environmental conditions. Sorghum is a cereal crop that exhibits exceptional tolerance to adverse conditions, in particular, water-limiting environments. This study utilized next generation sequencing (NGS) technology to examine the transcriptome of sorghum plants challenged with osmotic stress and exogenous abscisic acid (ABA) in order to elucidate genes and gene networks that contribute to sorghum's tolerance to water-limiting environments with a long-term aim of developing strategies to improve plant productivity under drought. RNA-Seq results revealed transcriptional activity of 28,335 unique genes from sorghum root and shoot tissues subjected to polyethylene glycol (PEG)-induced osmotic stress or exogenous ABA. Differential gene expression analyses in response to osmotic stress and ABA revealed a strong interplay among various metabolic pathways including abscisic acid and 13-lipoxygenase, salicylic acid, jasmonic acid, and plant defense pathways. Transcription factor analysis indicated that groups of genes may be co-regulated by similar regulatory sequences to which the expressed transcription factors bind. We successfully exploited the data presented here in conjunction with published transcriptome analyses for rice, maize, and Arabidopsis to discover more than 50 differentially expressed, drought-responsive gene orthologs for which no function had been previously ascribed. The present study provides an initial assemblage of sorghum genes and gene networks regulated by osmotic stress and hormonal treatment. We are providing an RNA-Seq data set and an initial collection of transcription factors, which offer a preliminary look into the cascade of global gene expression patterns that arise in a drought tolerant crop subjected to abiotic stress. These resources will allow scientists to query gene expression and functional annotation in response to drought.


Open multi-culture platform for simple and flexible study of multi-cell type interactions.
The study of multi-cell-type (MCT) interactions has the potential to significantly impact our understanding of tissue and disease biology. Such studies require innovative culture tools for unraveling the contributions of each cell type. Micro- and macro-scale platforms for MCT culture each have different advantages and disadvantages owing to their widely different capabilities, availability, and ease-of-use. However, as evidenced in the literature, there are very few examples of MCT studies and culture platforms, suggesting both biological and technical barriers. We have developed an open multi-culture platform to promote more rapid progress by integrating advantages of both micro- and macro-scale culture devices. The proposed open multi-culture platform addresses technical barriers by allowing easy customization, independent control of basic physical culture parameters, and incorporation of multiple culture modalities (e.g., 2D, 3D, transwell, and spheroid). The design also permits the user to obtain independent endpoints for each culture region. We demonstrate use of the platform in two example studies where we evaluated how cell ratio and cell types influence the response of triple negative breast cancer cells to heat damage and Hedgehog signaling. We also show that the platform can improve soluble factor transport between cell types compared to compartmentalized macro- and micro-scale alternatives. Last, we examine current and future challenges of the platform. We envision simple, yet flexible and customizable, platforms such as this will be important for advancing in vitro study of tissue and tumor biology.


Effects of sports activity in athletes with bicuspid aortic valve and mild aortic regurgitation.
In this study, the impact of regular training on left ventricle morphology in a group of athletes with bicuspid aortic valve (BAV) was evaluated. Longitudinal cohort study. A group of competitive athletes with BAV was followed up with a yearly standard echocardiographic examination for 5 years. Sport Medicine Centre, University of Florence, Pre-Participation Protocol Study. A group of 88 consecutive athletes diagnosed with BAV was identified in the period January to December 1999, and 30 of these completed a 5-year follow-up. They were compared with a group of 56 athletes with a normal tricuspid valve (TAV). BAV athletes showed significant progressive increase in left ventricular dimensions and aortic diameters at four levels. The values were within the range of the general and non-athletic BAV populations. In TAV athletes, the aortic and left ventricle dimensions did not increase significantly and remained within physiological range. Left ventricular dimensions in competitive BAV athletes remain within the normal range. There is, however, a significant progressive increase in the BAV group compared with the TAV group. These results are in agreement with data obtained in previous studies on the non-athletic BAV population. Sports activity does not have an additional effect on cardiac morphology in athletes with asymptomatic BAV associated with mild regurgitation, for at least 5 years.


PREDICTORS OF BIOCHEMICAL REMISSION AND RECURRENCE AFTER SURGICAL AND RADIATION TREATMENTS OF CUSHING DISEASE: A SYSTEMATIC REVIEW AND META-ANALYSIS.
We conducted a systematic review and meta-analysis to synthesize the evidence about predictors that may affect biochemical remission and recurrence after transsphenoidal surgery (TSS), radiosurgery (RS), and radiotherapy (RT) in Cushing disease. We searched multiple databases through December 2014 including original controlled and uncontrolled studies that enrolled patients with Cushing disease who received TSS (first-line), RS, or RT. We extracted data independently, in duplicates. Outcomes of interest were biochemical remission and recurrence. A meta-analysis was conducted using the random-effects model to estimate event rates with 95% confidence intervals (CIs). First-line TSS was associated with high remission (76% [95% CI, 72 to 79%]) and low recurrence rates (10% [95% CI, 6 to 16%]). Remission after TSS was higher in patients with microadenomas or positive-adrenocorticotropic hormone tumor histology. RT was associated with a high remission rate (RS, 68% [95% CI, 61 to 77%]; RT, 66% [95% CI, 58 to 75%]) but also with a high recurrence rate (RS, 32% [95% CI, 16 to 60%]; RT, 26% [95% CI, 14 to 48%]). Remission after RS was higher at short-term follow-up (≤2 years) and with high-dose radiation, while recurrence was higher in women and with lower-dose radiation. Remission was after RT in adults who received TSS prior to RT, and with lower radiation doses. There was heterogeneity (nonstandardization) in the criteria and cutoff points used to define biochemical remission and recurrence. First-line TSS is associated with high remission and low recurrence, while RS and RT are associated with reasonable remission rates but important recurrence rates. The current evidence warrants low confidence due to the noncomparative nature of the studies, high heterogeneity, and imprecision.


Clinico-hematological profile of Chediak-Higashi syndrome: experience from a tertiary care center in south India.
Chediak-Higashi syndrome (CHS) is a rare autosomal recessive disorder characterized by partial ocular and cutaneous albinism, increased susceptibility to pyogenic infections, the presence of large lysosomal-like organelles in most granule-containing cells and a bleeding tendency. The abnormal granules are most readily seen in blood and marrow leukocytes, especially granulocytes; and in melanocytes. Other clinical features include silvery hair, photophobia, horizontal and rotatory nystagmus and hepatosplenomegaly. The clinico-hematological profile of a series of 5 cases of CHS encountered at JIPMER Hospital with diagnostic work-up done in the Department of Pathology over the last 6 years is presented. The diagnostic work-up included complete hemogram with peripheral smear, bone marrow examination, skin and liver biopsies. The age of the patients ranged from 5 months to 3 years. All patients had silvery hair and partial albinism and presented with fever and recurrent chest infection. Two patients were stable. Three patients were in accelerated phase; of them, 1 patient with associated hemophagocytic syndrome had a rapidly fulminant course. Peripheral blood smear showed anomalously large granules in the leukocytes. Skin biopsy showed sparse, coarse melanin pigment in the epidermis, and liver biopsy done in 2 patients with accelerated phase showed portal lymphohistiocytic aggregates. The diagnostic hallmark of CHS is the occurrence of giant inclusion bodies (granules) in the peripheral leukocyte and their bone marrow precursors. The case series is being presented because of the rarity of CHS and varied spectrum of clinical and hematological presentation.


Microtubule polarity and distribution in teleost photoreceptors.
We have characterized the polarity orientation of microtubules in teleost retinal photoreceptors. The highly polarized rods and cones contain large numbers of paraxially aligned microtubules and exhibit dramatic cell shape changes. The myoid portion of the inner segments of both rods and cones undergoes contraction and elongation in response to light or circadian signals. Previous studies in our laboratory have demonstrated that in cones but not rods myoid elongation is microtubule-dependent. To determine polarity orientation, we decorated microtubules in photoreceptors of the green sunfish Lepomis cyanellus, with hooks formed from either exogenous or endogenous tubulin subunits. The direction of curvature of the attached hooks in cross section indicates microtubule polarity orientation by allowing one to determine the relative positions within the cell of the plus (fast-growing) and minus (slow-growing) ends of the microtubules. We found that virtually all cytoplasmic microtubules in photoreceptors are oriented with plus ends directed toward the synapse and minus ends toward the basal body at the base of the outer segment. Axonemal microtubules in photoreceptor outer segments are oriented with minus ends toward the basal body as in cilia and flagella. We have suggested previously that cone myoid elongation is mediated by mechanochemical sliding between microtubules. The polarity observations reported here indicate that if microtubules do slide in cones, sliding would necessarily occur between microtubules of parallel orientation as is observed in cilia and flagella.


How Severely Is DNA Quantification Hampered by RNA Co-extraction?
The optional RNase digest that is part of many DNA extraction protocols is often omitted, either because RNase is not provided in the kit or because users do not want to risk contaminating their laboratory. Consequently, co-eluting RNA can become a "contaminant" of unknown magnitude in a DNA extraction. We extracted DNA from liver, lung, kidney, and heart tissues and established that 28-52% of the "DNA" as assessed by spectrophotometry is actually RNA (depending on tissue type). Including an RNase digest in the extraction protocol reduced 260:280 purity ratios. Co-eluting RNA drives an overestimation of DNA yield when quantification is carried out using OD 260 nm spectrophotometry, or becomes an unquantified contaminant when spectrofluorometry is used for DNA quantification. This situation is potentially incompatible with the best practice guidelines for biobanks issued by organizations such as the International Society for Biological and Environmental Repositories, which state that biospecimens should be accurately characterized in terms of their identity, purity, concentration, and integrity. Consequently, we conclude that an RNase digest must be included in DNA extractions if pure DNA is required. We also discuss the implications of unquantified RNA contamination in DNA samples in the context of laboratory accreditation schemes.


The PAX2 tanscription factor is expressed in cystic and hyperproliferative dysplastic epithelia in human kidney malformations.
Human dysplastic kidneys are developmental aberrations which are responsible for many of the very young children with chronic renal failure. They contain poorly differentiated metanephric cells in addition to metaplastic elements. We recently demonstrated that apoptosis was prominent in undifferentiated cells around dysplastic tubules (Winyard, P.J.D., J. Nauta, D.S. Lirenman, P. Hardman, V.R. Sams, R.A. Risdon, and A.S. Woolf. 1996. Kidney Int. 49:135-146), perhaps explaining the tendency of some of these organs to regress. In contrast, apoptosis was rare in dysplastic epithelia which are thought to be ureteric bud malformations. On occasion, these tubules form cysts which distend the abdominal cavity (the multicystic dysplastic kidney) and dysplastic kidneys may rarely become malignant. We now demonstrate that dysplastic tubules maintain a high rate of proliferation postnatally and that PAX2, a potentially oncogenic transcription factor, is expressed in these epithelia. In contrast, both cell proliferation and PAX2 are downregulated during normal maturation of human collecting ducts. We demonstrate that BCL2, a protein which prevents apoptosis in renal mesenchymal to epithelia] conversion, is expressed ectopically in dysplastic kidney epithelia. We propose that dysplastic cyst formation may be understood in terms of aberrant temporal and spatial expression of master genes which are tightly regulated in the normal program of human nephrogenesis.


Manifesto for a European research network into obsessive-compulsive and related disorders.
Obsessive-compulsive and related disorders (O-CRDs) are highly disabling psychiatric illnesses of early-onset. They are responsible for considerable morbidity and socioeconomic burden. Existing treatments are usually only partially successful and there is an urgent need to understand the aetiological factors and neurobiological bases of the disorders in order to develop new and more effective strategies for prevention, early detection and effective treatment. Emerging data from the neurosciences supports the reconceptualisation of obsessive-compulsive disorder as a spectrum disorder, related to but different from the anxiety disorders and closely aligned with other less well understood psychiatric disorders characterised by compulsive acts such as body dysmorphic disorder, trichotillomania, skin-picking disorder, hoarding disorder; and possibly extending to tic disorders and other neurodevelopmental disorders such as autism. A new, O-CRDs research network, supported by the Networks Initiative of the European College of Neuropsychopharmacology and comprising leading figures in preclinical and clinical research, has been established. It aims to provide a European perspective on the current debate around internationally-accepted diagnostic criteria and treatment strategies for O-CRDs. Its objectives include; (1) identifying the key outstanding research questions that depend upon cross-centre collaborative investigation, (2) setting a research agenda that is likely to produce an impact on health-outcomes, and (3) strengthening existing projects and collaborative enterprises with these objectives in mind. This paper reviews some of these critical research priorities. By establishing shared multinational databases, collaborative research networks, multicentre studies and joint publications, it is hoped that progress will be achieved.


A multi-site study on medical school selection, performance, motivation and engagement.
Medical schools seek ways to improve their admissions strategies, since the available methods prove to be suboptimal for selecting the best and most motivated students. In this multi-site cross-sectional questionnaire study, we examined the value of (different) selection procedures compared to a weighted lottery procedure, which includes direct admission based on top pre-university grade point averages (≥8 out of 10; top-pu-GPA). We also considered whether students had participated in selection, prior to being admitted through weighted lottery. Year-1 (pre-clinical) and Year-4 (clinical) students completed standard validated questionnaires measuring quality of motivation (Academic Self-regulation Questionnaire), strength of motivation (Strength of Motivation for Medical School-Revised) and engagement (Utrecht Work Engagement Scale-Student). Performance data comprised GPA and course credits in Year-1 and clerkship performance in Year-4. Regression analyses were performed. The response rate was 35% (387 Year-1 and 273 Year-4 students). Top-pu-GPA students outperformed selected students. Selected Year-1 students reported higher strength of motivation than top-pu-GPA students. Selected students did not outperform or show better quality of motivation and engagement than lottery-admitted students. Participation in selection was associated with higher engagement and better clerkship performance in Year-4. GPA, course credits and strength of motivation in Year-1 differed between students admitted through different selection procedures. Top-pu-GPA students perform best in the medical study. The few and small differences found raise questions about the added value of an extensive selection procedure compared to a weighted lottery procedure. Findings have to be interpreted with caution because of a low response rate and small group sizes.


Synthesis of 2-hydroxy-1,4-oxazin-3-ones through ring transformation of 3-hydroxy-4-(1,2-dihydroxyethyl)-β-lactams and a study of their reactivity.
The reactivity of 3-hydroxy-4-(1,2-dihydroxyethyl)-β-lactams with regard to the oxidant sodium periodate was evaluated, unexpectedly resulting in the exclusive formation of new 2-hydroxy-1,4-oxazin-3-ones through a C3C4 bond cleavage of the intermediate 4-formyl-3-hydroxy-β-lactams followed by a ring expansion. This peculiar transformation stands in sharp contrast with the known NaIO(4)-mediated oxidation of 3-alkoxy- and 3-phenoxy-4-(1,2-dihydroxyethyl)-β-lactams, which exclusively leads to the corresponding 4-formyl-β-lactams without a subsequent ring enlargement. In addition, this new class of functionalized oxazin-3-ones was further evaluated for its potential use as building blocks in the synthesis of a variety of differently substituted oxazin-3-ones, morpholin-3-ones and pyrazinones. Furthermore, additional insights into the mechanism and the factors governing this new ring-expansion reaction were provided by means of density functional theory calculations.


Rapid neurogenesis through transcriptional activation in human stem cells.
Advances in cellular reprogramming and stem cell differentiation now enable ex vivo studies of human neuronal differentiation. However, it remains challenging to elucidate the underlying regulatory programs because differentiation protocols are laborious and often result in low neuron yields. Here, we overexpressed two Neurogenin transcription factors in human-induced pluripotent stem cells and obtained neurons with bipolar morphology in 4 days, at greater than 90% purity. The high purity enabled mRNA and microRNA expression profiling during neurogenesis, thus revealing the genetic programs involved in the rapid transition from stem cell to neuron. The resulting cells exhibited transcriptional, morphological and functional signatures of differentiated neurons, with greatest transcriptional similarity to prenatal human brain samples. Our analysis revealed a network of key transcription factors and microRNAs that promoted loss of pluripotency and rapid neurogenesis via progenitor states. Perturbations of key transcription factors affected homogeneity and phenotypic properties of the resulting neurons, suggesting that a systems-level view of the molecular biology of differentiation may guide subsequent manipulation of human stem cells to rapidly obtain diverse neuronal types.


CYP2E1--biochemical and toxicological aspects and role in alcohol-induced liver injury.
Ethanol-induced oxidative stress appears to play a major role in mechanisms by which ethanol causes liver injury. Many pathways have been suggested to contribute to the ability of ethanol to induce a state of oxidative stress. One central pathway appears to be the induction of the CYP2E1 form of cytochrome P450 enzymes by ethanol. CYP2E1 is of interest because of its ability to metabolize and activate many toxicological substrates, including ethanol, to more reactive toxic products. Levels of CYP2E1 are elevated under a variety of physiological and pathophysiological conditions, and after acute and chronic alcohol treatment. CYP2E1 is also an effective generator of reactive oxygen species such as the superoxide anion radical and hydrogen peroxide, and in the presence of iron catalysts, it produces powerful oxidants such as the hydroxyl radical. This review article summarizes some of the biochemical and toxicological properties of CYP2E1, and briefly describes the use of HepG2 cell lines developed to constitutively express the human CYP2E1 in assessing the actions of CYP2E1. Regulation of CYP2E1 is quite complex and will be briefly reviewed. Future directions in research, which may help clarify the actions of CYP2E1 and its role in alcoholic liver injury, are suggested.


Factor VIII: Long-established role in haemophilia A and emerging evidence beyond haemostasis.
Factor VIII protein (FVIII) as a coagulation replacement factor has for decades been used as the standard of care for management of people with haemophilia A. It is effective for treatment of bleeding events, as prophylaxis to prevent bleeding events and preserve joint function, and to support surgery in people with haemophilia A. Despite long experience in treating haemophilia A, we are only beginning to understand the functions of FVIII beyond its established role as a coenzyme to factor IXa to expedite thrombin generation through the intrinsic pathway of coagulation. Here, we review the current role of FVIII coagulant (FVIII:C) in haemophilia A management and emerging evidence for the role of FVIII across multiple systems, including the cardiovascular system, angiogenesis and maintenance of bone health. For instance, supraphysiological FVIII levels are a risk factor for venous thromboembolism. von Willebrand factor (VWF), which forms a non-covalent complex with circulating FVIII, is an established marker and regulator of angiogenesis. In a mouse model of haemophilia, treatment with FVIII decreased expression of receptor activator of nuclear factor kappa-Β ligand (RANKL), a marker for bone turnover. Longitudinal follow-up data in people with haemophilia A are needed to confirm and extend these observations.