Clinical trial metadata: defining and extracting metadata on the design, conduct, results and costs of 125 randomised clinical trials funded by the National Institute for Health Research Health Technology Assessment programme.
By 2011, the Health Technology Assessment (HTA) programme had published the results of over 100 trials with another 220 in progress. The aim of the project was to develop and pilot 'metadata' on clinical trials funded by the HTA programme. The aim of the project was to develop and pilot questions describing clinical trials funded by the HTA programme in terms of it meeting the needs of the NHS with scientifically robust studies. The objectives were to develop relevant classification systems and definitions for use in answering relevant questions and to assess their utility. Published monographs and internal HTA documents. A database was developed, 'populated' using retrospective data and used to answer questions under six prespecified themes. Questions were screened for feasibility in terms of data availability and/or ease of extraction. Answers were assessed by the authors in terms of completeness, success of the classification system used and resources required. Each question was scored to be retained, amended or dropped. One hundred and twenty-five randomised trials were included in the database from 109 monographs. Neither the International Standard Randomised Controlled Trial Number nor the term 'randomised trial' in the title proved a reliable way of identifying randomised trials. Only limited data were available on how the trials aimed to meet the needs of the NHS. Most trials were shown to follow their protocols but updates were often necessary as hardly any trials recruited as planned. Details were often lacking on planned statistical analyses, but we did not have access to the relevant statistical plans. Almost all the trials reported on cost-effectiveness, often in terms of both the primary outcome and quality-adjusted life-years. The cost of trials was shown to depend on the number of centres and the duration of the trial. Of the 78 questions explored, 61 were well answered, 33 fully with 28 requiring amendment were the analysis updated. The other 17 could not be answered with readily available data. The study was limited by being confined to 125 randomised trials by one funder. Metadata on randomised controlled trials can be expanded to include aspects of design, performance, results and costs. The HTA programme should continue and extend the work reported here. The National Institute for Health Research HTA programme.


The University of North Carolina's Health Care Pharmacy Assistance Progam.
UNC Health Care has a Pharmacy Assistance Program (PAP) that provides financial assistance to uninsured patients in North Carolina who need prescription medications. Despite significant investment in the program, little is known about the patients accessing it or the specific health care services they use. Our objectives were to describe the PAP population, to examine their use of prescription medications and health services, and to characterize changes in prescription medication use and expenditures from 2009 through 2011. We used a repeated cross-sectional study design, merging prescription claims with health records from Carolina Data Warehouse for Health, to measure use of prescription medications and use of inpatient, outpatient, and emergency department care by PAP participants. Prescription claims were grouped into therapeutic categories. We generated descriptive statistics for key variables to examine health service utilization from 2009 through 2011. From 2009 through 2011, PAP served 7,180 patients from 81 counties in North Carolina. PAP users received a mean of 23 prescriptions, at an average cost of $754 per recipient per year. An average of $2.93 million per year was spent on the program, with an 8% rise in spending from 2009 to 2011. Inpatient care and emergency department care were utilized by 30% and 31% of PAP users, respectively, and there was minimal change in these rates over 3 years. Data were limited to medications dispensed through PAP and to health services provided by UNC Health Care. With the state's decision to not expand Medicaid, PAP will continue to be an important resource for North Carolina's low income citizens.


Treatment of medulloblastoma with a modified measles virus.
Although treatment of medulloblastoma has improved, at least 30% of patients with this tumor die of progressive disease. Unfortunately, many of the children who survive suffer long-term treatment-related morbidity. Previous studies have demonstrated the efficacy of using oncolytic viruses to eradicate brain tumors. The objective of this study was to test the efficacy of measles virus in treating medulloblastoma. To determine whether medulloblastoma cells are susceptible, 5 different human medulloblastoma cell lines were analyzed for the expression of the measles virus receptor CD46. Fluorescence-activated cell-sorting analysis confirmed expression of CD46 on all cell lines tested, with UW288-1 having the most prominent expression and D283med displaying the lowest expression. CD46 expression was also demonstrated, using immunohistochemistry, in 13 of 13 medulloblastoma tissue specimens. All 5 medulloblastoma cell lines were examined for their susceptibility to measles virus killing in vitro. A measles virus containing the green fluorescent protein (GFP) gene as a marker for infection (MV-GFP) was used. All cell lines exhibited significant killing when infected with MV-GFP, all formed syncytia with infection, all showed fluorescence, and all allowed viral replicaton after infection. In an intracerebral murine xenograft model, a statistically significant increase in survival was seen in animals treated with the active measles virus compared with those treated with inactivated virus. These data demonstrate that medulloblastoma is susceptible to measles virus killing and that the virus may have a role in treating this tumor in the clinical setting.


Pulse shaping with birefringent crystals: a tool for quantum metrology.
A method for time differentiation based on a Babinet-Soleil-Bravais compensator is introduced. The complex transfer function of the device is measured using polarization spectral interferometry. Time differentiation of both the pulse field and pulse envelope are demonstrated over a spectral width of about 100 THz with a measured overlap with the objective mode greater than 99.8%. This pulse shaping technique is shown to be perfectly suited to time metrology at the quantum limit.


Impact of institute and person variables on teachers' conceptions of learning and teaching.
Teachers' conceptions of learning and teaching are important for faculty development to result in enduring changes in teaching practices. Until now, studies on these conceptions have mostly focused on traditional, lecture-based curricula rather than on small-group student-centred educational formats, which are gaining ground worldwide. To explore which factors predict teachers' conceptions in student-centred curricula. In two Dutch medical schools with 10 and 40 years of student-centred education, teachers were asked to fill out the Conceptions of Learning and Teaching (COLT) Questionnaire to assess their 'teacher-centredness', 'appreciation of active learning' and 'orientation to professional practice'. Next, we quantitatively assessed the relations of teachers' conceptions with their personal and occupational characteristics and institute. Overall response was 49.4% (N = 319/646). Institute was the main predictor for variance in all three scales, and discipline, gender and teaching experience significantly explained variance in two of the scales. More than 80% of the variance was not explained by these factors. Longer exposure to a student-centred curriculum was associated with fewer teacher-centred conceptions, greater 'appreciation of active learning' and stronger 'orientation towards professional practice'. In line with studies on lecture-based curricula, discipline, gender and teaching experience also appeared important for teachers' conceptions in student-centred curricula. More research is necessary to better understand the influence of institute on the three teachers' conceptions scales.


Tolerance induced by apoptotic antigen-coupled leukocytes is induced by PD-L1+ and IL-10-producing splenic macrophages and maintained by T regulatory cells.
Ag-specific tolerance is a highly desired therapy for immune-mediated diseases. Intravenous infusion of protein/peptide Ags linked to syngeneic splenic leukocytes with ethylene carbodiimide (Ag-coupled splenocytes [Ag-SP]) has been demonstrated to be a highly efficient method for inducing peripheral, Ag-specific T cell tolerance for treatment of autoimmune disease. However, little is understood about the mechanisms underlying this therapy. In this study, we show that apoptotic Ag-SP accumulate in the splenic marginal zone, where their uptake by F4/80(+) macrophages induces production of IL-10, which upregulates the expression of the immunomodulatory costimulatory molecule PD-L1 that is essential for Ag-SP tolerance induction. Ag-SP infusion also induces T regulatory cells that are dispensable for tolerance induction but required for long-term tolerance maintenance. Collectively, these results indicate that Ag-SP tolerance recapitulates how tolerance is normally maintained in the hematopoietic compartment and highlight the interplay between the innate and adaptive immune systems in the induction of Ag-SP tolerance. To our knowledge, we show for the first time that tolerance results from the synergistic effects of two distinct mechanisms, PD-L1-dependent T cell-intrinsic unresponsiveness and the activation of T regulatory cells. These findings are particularly relevant as this tolerance protocol is currently being tested in a Phase I/IIa clinical trial in new-onset relapsing-remitting multiple sclerosis.


Prioritising physical and psychological symptoms: what are the barriers and facilitators to the discussion of anxiety in the primary care consultation?
Anxiety is under-recorded and under-treated in the UK and is under-represented in research compared with depression. Detecting anxiety can be difficult because of co-existing conditions. GPs can be reluctant to medicalise anxiety symptoms and patients can be reluctant to disclose them, for a variety of reasons. This research addresses the gap in evidence of real-life consultations of patients with anxiety and explores how physical and psychological symptoms are discussed and prioritised by patients and GPs in primary care consultations. A mixed methods study using a baseline questionnaire, video-recorded primary care consultations and interview data with patients and GPs. Seventeen patients with anxiety symptoms (GAD-7 score ≥ 10) completed a questionnaire, had their consultation video-recorded and took part in a semi-structured interview. Four GPs were interviewed. The main themes that emerged from GP and patients accounts as barriers and facilitators to discussing anxiety mostly mirrored each other. The GP/patient relationship and continuity of care was the main facilitator for the discussion of anxiety in the consultation. The main barriers were: attribution of or unacknowledged symptoms; co-morbidities; and time constraints. GPs overcame these barriers by making repeat appointments and employing prioritising techniques; patients by choosing an empathetic GP. The findings add to the evidence base concerning the management of anxiety in primary care. The findings suggest that the discussion around anxiety is a process negotiated between the patient and the GP influenced by a range of barriers and facilitators. Co-existing depression and health anxieties can mask anxiety symptoms in patients. Good practice techniques such as bringing back patients for appointments to foster continuity of care and understanding can help disclosure and detection of anxiety symptoms. Future research could investigate this longitudinally and should include a wider range of GPs practices and GPs.


Characterization of viability and proliferation of alginate-poly-L-lysine-alginate encapsulated myoblasts using flow cytometry.
Genetically modified cells encapsulated in alginate-poly-L-lysine-alginate (APA) are being developed to deliver therapeutic products to treat a variety of diseases. The characterization of the encapsulated cells thus becomes paramount. This study reports a novel method to assess the viability, granularity and proliferation of encapsulated cells based on flow cytometry. The in vitro viability of encapsulated G8 murine myoblasts secreting canine FVIII (cFVIII) measured by flow cytometry was comparable to the traditional trypan blue exclusion method and both correlated with cFVIII secretion levels. In contrast, after implantation into mice, only viability measured by flow cytometry correlated with cFVIII secretion. Further, flow cytometry analysis of encapsulated cells maintained in vitro and in vivo revealed a greater fraction of granular cells compared to free cells, suggesting that encapsulation influences the morphology (cytoplasmic composition) of cells within APA microcapsules. Interestingly, the proliferation study showed that encapsulated cells proliferate faster, on average, and were more heterogeneous in vivo compared to in vitro culture conditions, suggesting that encapsulated cell proliferation is complex and environment-dependent. In conclusion, we show that flow cytometry analysis allows for a more consistent and comprehensive examination of encapsulated cells to aid in the development of cell therapy protocols.


Shedding of the tumor necrosis factor (TNF) receptor from the surface of hepatocytes during sepsis limits inflammation through cGMP signaling.
Proteolytic cleavage of the tumor necrosis factor (TNF) receptor (TNFR) from the cell surface contributes to anti-inflammatory responses and may be beneficial in reducing the excessive inflammation associated with multiple organ failure and mortality during sepsis. Using a clinically relevant mouse model of polymicrobial abdominal sepsis, we found that the production of inducible nitric oxide synthase (iNOS) in hepatocytes led to the cyclic guanosine monophosphate (cGMP)-dependent activation of the protease TACE (TNF-converting enzyme) and the shedding of TNFR. Furthermore, treating mice with a cGMP analog after the induction of sepsis increased TNFR shedding and decreased systemic inflammation. Similarly, increasing the abundance of cGMP with a clinically approved phosphodiesterase 5 inhibitor (sildenafil) also decreased markers of systemic inflammation, protected against organ injury, and increased circulating amounts of TNFR1 in mice with sepsis. We further confirmed that a similar iNOS-cGMP-TACE pathway was required for TNFR1 shedding by human hepatocytes in response to the bacterial product lipopolysaccharide. Our data suggest that increasing the bioavailability of cGMP might be beneficial in ameliorating the inflammation associated with sepsis.


Cymonomid crabs of the MAINBAZA Expedition (Decapoda: Brachyura).
Cymonomid crabs collected from the Mozambique Channel off Madagascar by the 2011 MAINBAZA Expedition are reported. Two species of Cymonomus A. Milne Edwards, 1880, are represented, of which one is new to science and the other, C. valdiviae Lankester, 1903, is rediscovered, being previously known only from the holotype. Three species of Cymonomidae are now known from the western Indian Ocean, including C. trifurcus Stebbing, 1920, from South Africa.


Evidence of unmet need in the care of severely physically disabled adults.
To identify unmet needs in the care of severely disabled people aged 16-64. Detailed personal interview and physical assessment of physically disabled adults; personal or telephone interview with carers. Somerset Health District. 181 severely disabled adults and their carers. Independence in activities of daily living; identity of requirements for assessing communication disorders; appropriate provision of services and allowances. 53 (29.3%) of the 181 disabled subjects had unmet needs for aids to allow independence in activities of daily living-namely, 43% of subjects (41/95) with progressive disorders and 14% of subjects (12/86) with non-progressive disorders. The prevalence of unmet need was higher among subjects whose sole regular professional contact was with health services personnel (48 (40.3%) of 119 subjects). Only 18 (31.6%) of the 57 subjects with communication disorders had ever been assessed by a speech therapist. This study shows that the needs of severely physically disabled adults in the community--especially those with progressive disorders--are being monitored inadequately by health professionals.


Determination of the real effect of genes identified in GWAS: the example of IL2RA in multiple sclerosis.
Genome-wide association studies (GWAS), although efficient to detect genes involved in complex diseases, are not designed to measure the real effect of the genes. This is illustrated here by the example of IL2RA in multiple sclerosis (MS). Association between IL2RA and MS is clearly established, although the functional variation is still unknown: the effect of IL2RA might be better described by several SNPs than by a single one. This study investigates whether a pair of SNPs better explains the observed linkage and association data than a single SNP. In total, 522 trio families and 244 affected sib-pairs were typed for 26 IL2RA SNPs. For each SNP and pairs of SNPs, the phased genotypes of patients and controls were compared to determine the SNP set offering the best risk discrimination. Consistency between the genotype risks provided by the retained set and the identical by descent allele sharing in affected sib-pairs was assessed. After controlling for multiple testing, the set of SNPs rs2256774 and rs3118470, provides the best discrimination between the case and control genotype distributions (P-corrected=0.009). The relative risk between the least and most at-risk genotypes is 3.54 with a 95% confidence interval of [2.14-5.94]. Furthermore, the linkage information provided by the allele sharing between affected sibs is consistent with the retained set (P=0.80) but rejects the SNP reported in the literature (P=0.006). Establishing a valid modeling of a disease gene is essential to test its potential interaction with other genes and to reconstruct the pathophysiological pathways.


Dioxin-like PCBs and endometriosis.
A recent survey in the United States identified 287 different chemicals in human cord blood, demonstrating the significant exposure of women and their children to a wide array of environmental toxicants. While reducing contamination and exposure should be an international priority, it is equally appropriate to develop an understanding of the health consequences of increasing world-wide industrialization. Endometriosis, a disease of the female reproductive tract, has emerged as a disease potentially related to environmental exposures. While a number of population-based studies have suggested that a woman's exposure to dioxin-like polychlorinated biphenyls may affect her risk of developing this disease, other studies have failed to find such evidence. In the current manuscript, we will review the limited data regarding polychlorinated biphenyl congeners and endometriosis with a focus on dioxin-like toxicants. We will also discuss the potential importance of early life exposures to these toxicants on the subsequent development of endometriosis.


Alkaloid variation among epichloid endophytes of sleepygrass (Achnatherum robustum) and consequences for resistance to insect herbivores.
Epichloid endophytes are well known symbionts of many cool-season grasses that may alleviate environmental stresses for their hosts. For example, endophytes produce alkaloid compounds that may be toxic to invertebrate or vertebrate herbivores. Achnatherum robustum, commonly called sleepygrass, was aptly named due to the presence of an endophyte that causes toxic effects to livestock and wildlife. Variation in alkaloid production observed in two A. robustum populations located near Weed and Cloudcroft in the Lincoln National Forest, New Mexico, suggests two different endophyte species are present in these populations. Genetic analyses of endophyte-infected samples revealed major differences in the endophyte alkaloid genetic profiles from the two populations, which were supported with chemical analyses. The endophyte present in the Weed population was shown to produce chanoclavine I, paspaline, and terpendoles, so thus resembles the previously described Epichloë funkii. The endophyte present in the Cloudcroft population produces chanoclavineI, ergonovine, lysergic acid amide, and paspaline, and is an undescribed endophyte species. We observed very low survival rates for aphids feeding on plants infected with the Cloudcroft endophyte, while aphid survival was better on endophyte infected plants in the Weed population. This observation led to the hypothesis that the alkaloid ergonovine is responsible for aphid mortality. Direct testing of aphid survival on oat leaves supplemented with ergonovine provided supporting evidence for this hypothesis. The results of this study suggest that alkaloids produced by the Cloudcroft endophyte, specifically ergonovine, have insecticidal properties.


Quantifying the infectivity of human immunodeficiency virus.
We have developed a mathematical model that quantifies lymphocyte infection by human immunodeficiency virus (HIV) and lymphocyte protection by blocking agents such as soluble CD4. We use this model to suggest standardized parameters for quantifying viral infectivity and to suggest techniques for calculating these parameters from well-mixed infectivity assays. We discuss the implications of the model for our understanding of the infectious process and virulence of HIV in vivo.


Oral Health Literacy Assessment: development of an oral health literacy instrument for Spanish speakers.
To develop an oral health literacy instrument for Spanish-speaking adults, evaluate its psychometric properties, and determine its comparability to an English version. The Oral Health Literacy Assessment in Spanish (OHLA-S) and English (OHLA-E) are designed with a word recognition section and a comprehension section using the multiple-choice format developed by an expert panel. Validation of OHLA-S and OHLA-E involved comparing the instrument with other health literacy instruments in a sample of 201 Spanish-speaking and 204 English-speaking subjects. Comparability between Spanish and English versions was assessed by testing for differential item functioning (DIF) using item response theory. We considered three OHLA-S scoring systems. Based on validity and reliability comparisons, 24 items were retained in the OHLA-S instrument. OHLA-S was correlated with another health literacy instrument, Spanish Test of Functional Health Literacy in Adults (P < 0.05). Significant correlations were also found between OHLA-S and years of schooling, oral health knowledge, overall health, and an understanding of written health-care materials (P < 0.05). OHLA-S displayed satisfactory reliability with a Cronbach Alpha of 0.70-0.80. DIF results suggested that OHLA-S and OHLA-E scores were not comparable at a given level of oral health literacy. OHLA-S has acceptable reliability and validity. OHLA-S and OHLA-E are two different measurement tools and should not be used to compare oral health literacy between English- and Spanish-speaking populations.


Markov models for biogeography-based optimization.
Biogeography-based optimization (BBO) is a population-based evolutionary algorithm that is based on the mathematics of biogeography. Biogeography is the science and study of the geographical distribution of biological organisms. In BBO, problem solutions are analogous to islands, and the sharing of features between solutions is analogous to the migration of species. This paper derives Markov models for BBO with selection, migration, and mutation operators. Our models give the theoretically exact limiting probabilities for each possible population distribution for a given problem. We provide simulation results to confirm the Markov models.


Improved precision in calculated T1 MR images using multiple spin-echo acquisition.
Calculated T1 images require that magnetic resonance signals be detected at several inversion or repetition times (TR). Multiple spin-echo (SE) acquisitions provide several measurements of the magnetization at each TR, the signal size diminishing according to T2 decay. In this work we review one method (Case 1) for estimating T1 from single echoes and present four new methods (Cases 2-5) in which multiple acquired echoes are used. For Case 2 a fit is performed using the first echo at each TR, repeated using second echoes, etc., and the final T1 estimate is the simple average of the individual fits at each echo time (TE). For Case 3 the optimum weighted average is performed. For Cases 4 and 5 synthetic SE images are generated at each TR prior to the T1 fit, Case 4 using a synthetic TE of zero, and Case 5 using a TE providing maximum signal-to-noise ratio in the synthetic image. The relative precision in T1 provided by each method is calculated rigorously. It is proven that Cases 3 and 5 are optimum and equivalent and can theoretically reduce the noise in T1 images by as much as 40% over Case 1 with no increase in scanning time. Approximations are proposed that enable the optimum methods to be implemented in a practical fashion. Experimental images are presented that verify the relative predicted behavior.


Weapon Carrying and Weapon Brandishing at School: Responses to Strain Among Same-Sex Attracted Youth.
Studies consistently find lesbian, gay, bisexual, and questioning youth are more likely than other youth to experience discrimination, fear for their safety, and be victimized at school. We argue that same-sex attracted youth may be more likely to both carry weapons and to brandish those weapons as coping mechanisms in response to the strain of real or perceived mistreatment school. Using data from the National Longitudinal Study of Adolescent to Adult Health (Wave 1, 1994-1995), we examine the relationships between same-sex attraction, fear, victimization, weapon carrying, and weapon brandishing at school. We find that same-sex attracted youth are significantly more likely than other youth to carry a weapon to school, and to brandish the weapon in front of peers. Consistent with general strain explanations, victimization, discrimination, and fear mediate the association between sexual minority status and both outcomes. We discuss the implications of these patterns for primary prevention interventions.


Cancer: beyond speciation.
A good account of the nature of cancer should provide not only a description of its consistent features, but also how they arise, how they are maintained, why conventional chemotherapy succeeds, and fails, and where to look for better targets. Cancer was once regarded as enigmatic and inexplicable; more recently, the "mutation theory," based on random alterations in a relatively small set of proto-oncogenes and tumor suppressor genes, has enjoyed widespread acceptance. The "mutation theory," however, is noticeable for its failure to explain the basis of differential chemosensitivity, for providing a paucity of targets, especially druggable ones, and for justifying the development of targeted therapies with, in general, disappointingly abbreviated clinical benefit. Furthermore, this theory has mistakenly predicted a widespread commonality of consistent genetic abnormalities across the range of cancers, whereas the opposite, that is, roiling macrogenomic instability, is generally the rule. In contrast, concerning what actually is consistent, that is, the suite of metabolic derangements common to virtually all, especially aggressive, cancers, the "Mutation Theory" has nothing to say. Other hypotheses merit serious consideration "aneuploidy theories" posit whole-genome instability and imbalance as causally responsible for the propagation of the tumor. Another approach, that is, "derepression atavism," suggests cancer results from the release of an ancient survival program, characterized by the emergence of remarkably primitive features such as unicellularity, fermentation, and immortality; existential goals are served by heuristic genomic instability coupled with host-to-tumor biomass interconversion, mediated by the Warburg effect, a major component of the program. Carcinogenesis is here seen as a process of de-speciation; however, genomic nonrestabilization raises issues as to where on the tree of life cancers belong, as a genuinely alternative modus vivendi. Philosophical considerations aside, genomic instability offers the prospect of subtle new therapies based on loss of information rather than gain; and the consistent, specific, and broad-spectrum perfidy of the Warburg effect highlights a supplemental target of the highest priority.


Corticotropin-releasing factor overexpression in mice abrogates sex differences in body weight, visceral fat, and food intake response to a fast and alters levels of feeding regulatory hormones.
Corticotropin-releasing factor overexpressing (CRF-OE) male mice showed an inhibited feeding response to a fast, and lower plasma acyl ghrelin and Fos expression in the arcuate nucleus compared to wild-type (WT) mice. We investigated whether hormones and hypothalamic feeding signals are impaired in CRF-OE mice and the influence of sex. Male and female CRF-OE mice and WT littermates (4-6 months old) fed ad libitum or overnight fasted were assessed for body, adrenal glands and perigonadal fat weights, food intake, plasma hormones, blood glucose, and mRNA hypothalamic signals. Under fed conditions, compared to WT, CRF-OE mice have increased adrenal glands and perigonadal fat weight, plasma corticosterone, leptin and insulin, and hypothalamic leptin receptor and decreased plasma acyl ghrelin. Compared to male, female WT mice have lower body and perigonadal fat and plasma leptin but higher adrenal glands weights. CRF-OE mice lost these sex differences except for the adrenals. Male CRF-OE and WT mice did not differ in hypothalamic expression of neuropeptide Y (NPY) and proopiomelanocortin (POMC), while female CRF-OE compared to female WT and male CRF-OE had higher NPY mRNA levels. After fasting, female WT mice lost more body weight and ate more food than male WT, while CRF-OE mice had reduced body weight loss and inhibited food intake without sex difference. In male WT mice, fasting reduced plasma insulin and leptin and increased acyl ghrelin and corticosterone while female WT showed only a rise in corticosterone. In CRF-OE mice, fasting reduced insulin while leptin, acyl ghrelin and corticosterone were unchanged with no sex difference. Fasting blood glucose was higher in CRF-OE with female > male. In WT mice, fasting increased hypothalamic NPY expression in both sexes and decreased POMC only in males, while in CRF-OE mice, NPY did not change, and POMC decreased in males and increased in females. These data indicate that CRF-OE mice have abnormal basal and fasting circulating hormones and hypothalamic feeding-related signals. CRF-OE also abolishes the sex difference in body weight, abdominal fat, and fasting-induced feeding and changes in plasma levels of leptin and acyl ghrelin.


Out-of-pocket healthcare payments on chronic conditions impoverish urban poor in Bangalore, India.
The burden of chronic conditions is on the rise in India, necessitating long-term support from healthcare services. Healthcare, in India, is primarily financed through out-of-pocket payments by households. Considering scarce evidence available from India, our study investigates whether and how out-of-pocket payments for outpatient care affect individuals with chronic conditions. A large census covering 9299 households was conducted in Bangalore, India. Of these, 3202 households that reported presence of chronic condition were further analysed. Data was collected using a structured household-level questionnaire. Out-of-pocket payments, catastrophic healthcare expenditure, and the resultant impoverishment were measured using a standard technique. The response rate for the census was 98.5%. Overall, 69.6% (95%CI=68.0-71.2) of households made out-of-pocket payments for outpatient care spending a median of 3.2% (95%CI=3.0-3.4) of their total income. Overall, 16% (95%CI=14.8-17.3) of households suffered financial catastrophe by spending more than 10% of household income on outpatient care. Occurrence and intensity of financial catastrophe were inequitably high among poor. Low household income, use of referral hospitals as place for consultation, and small household size were associated with a greater likelihood of incurring financial catastrophe.The out-of-pocket spending on chronic conditions doubled the number of people living below the poverty line in one month, with further deepening of their poverty. In order to cope, households borrowed money (4.2% instances), and sold or mortgaged their assets (0.4% instances). This study provides evidence from India that the out-of-pocket payment for chronic conditions, even for outpatient care, pushes people into poverty. Our findings suggest that improving availability of affordable medications and diagnostics for chronic conditions, as well as strengthening the gate keeping function of the primary care services are important measures to enhance financial protection for urban poor. Our findings call for inclusion of outpatient care for chronic conditions in existing government-initiated health insurance schemes.


Pristine C(60) fullerenes inhibit the rate of tumor growth and metastasis.
To estimate the impact of C(60) fullerene aqueous solution (C(60)FAS) on the rate of transplanted malignant tumor growth and metastasis. Lewis lung carcinoma was transplanted into С57Bl/6J male mice. Conventional methods for the evaluation of antitumor and antimetastatic effects have been used. The C(60)FAS at low single therapeutic dose of 5 mg/kg inhibited the growth of transplanted malignant tumor (antitumor effect) and metastasis (antimetastatic effect): the maximum therapeutic effect was found to be of 76.5% for the tumor growth inhibition; the increase of animal life span by 22% was found; the metastasis inhibition index was estimated as 48%. It was found that water-soluble pristine С(60) fullerenes efficiently inhibit the transplanted malignant tumor growth and metastasis.


Optimizing microsurgical skills with EEG neurofeedback.
By enabling individuals to self-regulate their brainwave activity in the field of optimal performance in healthy individuals, neurofeedback has been found to improve cognitive and artistic performance. Here we assessed whether two distinct EEG neurofeedback protocols could develop surgical skill, given the important role this skill plays in medicine. National Health Service trainee ophthalmic microsurgeons (N = 20) were randomly assigned to either Sensory Motor Rhythm-Theta (SMR) or Alpha-Theta (AT) groups, a randomized subset of which were also part of a wait-list 'no-treatment' control group (N = 8). Neurofeedback groups received eight 30-minute sessions of EEG training. Pre-post assessment included a skills lab surgical procedure with timed measures and expert ratings from video-recordings by consultant surgeons, together with state/trait anxiety self-reports. SMR training demonstrated advantages absent in the control group, with improvements in surgical skill according to 1) the expert ratings: overall technique (d = 0.6, p < 0.03) and suture task (d = 0.9, p < 0.02) (judges' intraclass correlation coefficient = 0.85); and 2) with overall time on task (d = 0.5, p = 0.02), while everyday anxiety (trait) decreased (d = 0.5, p < 0.02). Importantly the decrease in surgical task time was strongly associated with SMR EEG training changes (p < 0.01), especially with continued reduction of theta (4-7 Hz) power. AT training produced marginal improvements in technique and overall performance time, which were accompanied by a standard error indicative of large individual differences. Notwithstanding, successful within session elevation of the theta-alpha ratio correlated positively with improvements in overall technique (r = 0.64, p = 0.047). SMR-Theta neurofeedback training provided significant improvement in surgical technique whilst considerably reducing time on task by 26%. There was also evidence that AT training marginally reduced total surgery time, despite suboptimal training efficacies. Overall, the data set provides encouraging evidence of optimised learning of a complex medical specialty via neurofeedback training.


A novel Sarcocystis neurona genotype XIII is associated with severe encephalitis in an unexpectedly broad range of marine mammals from the northeastern Pacific Ocean.
Sarcocystis neurona is an important cause of protozoal encephalitis among marine mammals in the northeastern Pacific Ocean. To characterise the genetic type of S. neurona in this region, samples from 227 stranded marine mammals, most with clinical or pathological evidence of protozoal disease, were tested for the presence of coccidian parasites using a nested PCR assay. The frequency of S. neurona infection was 60% (136/227) among pinnipeds and cetaceans, including seven marine mammal species not previously known to be susceptible to infection by this parasite. Eight S. neurona fetal infections identified this coccidian parasite as capable of being transmitted transplacentally. Thirty-seven S. neurona-positive samples were multilocus sequence genotyped using three genetic markers: SnSAG1-5-6, SnSAG3 and SnSAG4. A novel genotype, referred to as Type XIII within the S. neurona population genetic structure, has emerged recently in the northeastern Pacific Ocean and is significantly associated with an increased severity of protozoal encephalitis and mortality among multiple stranded marine mammal species.


A recurrent germline mutation in the PIGA gene causes Simpson-Golabi-Behmel syndrome type 2.
Hypomorphic germline mutations in the PIGA (phosphatidylinositol glycan class A) gene recently were recognized as the cause of a clinically heterogeneous spectrum of X-linked disorders including (i) early onset epileptic encephalopathy with severe muscular hypotonia, dysmorphism, multiple congenital anomalies, and early death ("MCAHS2"), (ii) neurodegenerative encephalopathy with systemic iron overload (ferro-cerebro-cutaneous syndrome, "FCCS"), and (iii) intellectual disability and seizures without dysmorphism. Previous studies showed that the recurrent PIGA germline mutation c.1234C>T (p.Arg412*) leads to a clinical phenotype at the most severe end of the spectrum associated with early infantile lethality. We identified three additional individuals from two unrelated families with the same PIGA mutation. Major clinical findings include early onset intractable epileptic encephalopathy with a burst-suppression pattern on EEG, generalized muscular hypotonia, structural brain abnormalities, macrocephaly and increased birth weight, joint contractures, coarse facial features, widely spaced eyes, a short nose with anteverted nares, gingival overgrowth, a wide mouth, short limbs with short distal phalanges, and a small penis. Based on the phenotypic overlap with Simpson-Golabi-Behmel syndrome type 2 (SGBS2), we hypothesized that both disorders might have the same underlying cause. We were able to confirm the same c.1234C>T (p.Arg412*) mutation in the DNA sample from an affected fetus of the original family affected with SGBS2. We conclude that the recurrent PIGA germline mutation c.1234C>T leads to a recognizable clinical phenotype with a poor prognosis and is the cause of SGBS2.


From enzymatic adaptation to allosteric transitions.
The research by my collaborators and myself since 1945 has been carried out entirely at the Pasteur Institute. This work has received decisive assistance from numerous institutions, in particular the Centre National de la Recherche Scientifique, the Rockefeller Foundation of New York, the National Science Foundation and the National Institutes of Health of the United States, the Jane Coffin Childs Memorial Fund, the Commissariat á l'Energie Atomique, and the Depsilonlepsilongation Gepsilonnepsilonrale alpha la Recherche Scientifique et Technique. A donation by MIesdames Edouard de Rothschild and Bethsabee de Rothschild permitted, in large part, the establishment in 1954 of the Department of Cellular Biochemistry at the Pasteur Institute


A High Malaria Prevalence Identified by PCR among Patients with Acute Undifferentiated Fever in India.
Approximately one million malaria cases were reported in India in 2015, based on microscopy. This study aims to assess the malaria prevalence among hospitalised fever patients in India identified by PCR, and to evaluate the performance of routine diagnostic methods. During June 2011-December 2012, patients admitted with acute undifferentiated fever to seven secondary level community hospitals in Assam (Tezpur), Bihar (Raxaul), Chhattisgarh (Mungeli), Maharashtra (Ratnagiri), Andhra Pradesh (Anantapur) and Tamil Nadu (Oddanchatram and Ambur) were included. The malaria prevalence was assessed by polymerase chain reaction (PCR), routine microscopy, and a rapid diagnostic test (RDT) with PCR as a reference method. The malaria prevalence by PCR was 19% (268/1412) ranging from 6% (Oddanchatram, South India) to 35% (Ratnagiri, West India). Among malaria positive patients P. falciparum single infection was detected in 46%, while 38% had P. vivax, 11% mixed infections with P. falciparum and P. vivax, and 5% P. malariae. Compared to PCR, microscopy had sensitivity of 29% and specificity of 98%, while the RDT had sensitivity of 24% and specificity of 99%. High malaria prevalence was identified by PCR in this cohort. Routine diagnostic methods had low sensitivity compared to PCR. The results suggest that malaria is underdiagnosed in rural India. However, low parasitaemia controlled by immunity may constitute a proportion of PCR positive cases, which calls for awareness of the fact that other pathogens could be responsible for the febrile disease in submicroscopic malaria.


Hypothesis: Obesity Is Associated with a Lower Mutation Threshold in Colon Cancer.
Neoplastic progression requires accumulation of several mutations (mutation threshold). We hypothesize that obesity raises the risk of microsatellite stable (MSS) colon cancer (CC) at least in part by decreasing the mutation threshold. Thus, we posit that obese patients require fewer mutations, particularly driver mutations, compared to their normal BMI counterparts. Further, we suggest that the reduced number of required mutations in obese patients could be due to several factors, including the high levels of cytokines that accompany obesity. Cytokine-activated ERK, AKT, and JAK/STAT signaling could synergize with CC-initiating mutations to promote intestinal neoplastic development. Therefore, driver mutations that induce these specific pathways may not be "required" for neoplastic development in obesity; alteration in cell signaling consequent to obesity can substitute for some driver mutations in neoplastic progression. This hypothesis is supported by preliminary analyses of data from The Cancer Genome Atlas (TCGA). Thus, we observed that, compared to normal weight patients, cancer genomes of obese MSS CC patients exhibit fewer somatic mutations, and correspondingly lower numbers of mutations in driver genes (P = 0.026).The most striking observation was the lower number of KRAS mutations detected in patients with high body-mass index (BMI). These intriguing observations require further validation with increased number of patients, taking into account all possible confounding factors. If the hypothesis is confirmed, future studies should also address several possible explanations for the observed lower mutation threshold in obese MSS CC patients.


Parafalcine and midline arteriovenous malformations: surgical strategy, techniques, and outcomes.
Parafalcine arteriovenous malformations (AVMs) have a midline plane in common, but differ in their location (anterior, middle, or posterior) and depth (superficial or deep). Surgical management varies with AVM location and depth in terms of patient position, head position, craniotomy, and surgical approach. This study examined surgical strategies, patient outcomes, and regional factors influencing results. Patients with AVMs located on the medial surface of the cerebral hemisphere were identified retrospectively from a consecutive, single-neurosurgeon series that is registered prospectively as part of the UCSF Brain Arteriovenous Malformation Study Project. During a 12-year period, 443 patients with AVMs were treated surgically. Of these 443 patients, 132 (30%) had parafalcine AVMs, which were distributed in zones as follows: superficial-anterior, 25 (18.9%); superficial-middle, 26 (19.7%); superficial-posterior, 39 (29.5%); deep-anterior, 25 (18.9%); deep-posterior, 17 (12.9%). Five different surgical strategies were used depending on AVM zone. Complete AVM resection was achieved in 123 (93.2%) of 132 patients. Overall, neurological condition improved in 74 patients (56.1%) and remained unchanged in 41 patients (31.1%). Neurological condition deteriorated in 12 patients (9.1%), and 5 patients (3.8%) died. Patients with AVMs in the superficial-middle zone had the highest rate of neurological deterioration (26.9%). Parafalcine AVMs lie on a midline surface that, when exposed with a bilateral craniotomy across the superior sagittal sinus and a wide opening of the interhemispheric fissure, makes them superficial. However, unlike convexity AVMs, which are approached perpendicularly, parafalcine AVMs are approached tangentially. Gravity retraction is useful with deeply located AVMs (those in the deep-anterior and deep-posterior zones), because it widens the interhemispheric fissure and accesses deep arterial feeding vessels from the anterior and posterior cerebral arteries. Surgical risks were increased in the superficial-middle zone, which is likely explained by the proximity of sensorimotor cortex. The authors' regional classification of parafalcine AVMs may serve as a guide to surgical planning.


Protective effects of hydrogen-rich saline on monocrotaline-induced pulmonary hypertension in a rat model.
Hydrogen-rich saline has been reported to have antioxidant and anti-inflammatory effects and effectively protect against organ damage. Oxidative stress and inflammation contribute to the pathogenesis and/or development of pulmonary hypertension. In this study, we investigated the effects of hydrogen-rich saline on the prevention of pulmonary hypertension induced by monocrotaline in a rat model. In male Sprague-Dawley rats, pulmonary hypertension was induced by subcutaneous administration of monocrotaline at a concentration of 6 mg/100 g body weight. Hydrogen-rich saline (5 ml/kg) or saline was administered intraperitoneally once daily for 2 or 3 weeks. Severity of pulmonary hypertension was assessed by hemodynamic index and histologic analysis. Malondialdehyde and 8-hydroxy-desoxyguanosine level, and superoxide dismutase activity were measured in the lung tissue and serum. Levels of pro-inflammatory cytokines (tumor necrosis factor-α, interleukin-6) in serum were determined with enzyme-linked immunosorbent assay. Hydrogen-rich saline treatment improved hemodynamics and reversed right ventricular hypertrophy. It also decreased malondialdehyde and 8-hydroxy-desoxyguanosine levels, and increased superoxide dismutase activity in the lung tissue and serum, accompanied by a decrease in pro-inflammatory cytokines. These results suggest that hydrogen-rich saline ameliorates the progression of pulmonary hypertension induced by monocrotaline in rats, which may be associated with its antioxidant and anti-inflammatory effects.


[The use of propolis-based therapy in otorhionolaryngology].
The authors analyze the literature publications and the results of their original investigations concerning the application of the propolis-based pharmacological products for the treatment of acute and chronic diseases of the upper respiratory tract and ears. The desirability of the more extensive application of the propolis-containing medicines in the routine otorhinolaryngological practice is emphasized bearing in mind their numerous biological effects including antibiotic, anti-inflammatory, immunostimulating, anti-oxidative, anesthetic, and cytostatic actions.


Virulence of Pythium Species Isolated from Wheat Fields in Eastern Washington.
Although Pythium root rot in wheat (Triticum aestivum) is well documented, limited information is available concerning which species of Pythium are most responsible for disease damage. The objective of this study was to examine the variation in virulence on wheat among isolates of Pythium collected from cereal grain fields in eastern Washington. Isolates of nine Pythium species were tested for virulence on spring wheat cultivars Chinese Spring and Spillman. Cultivars were planted in pasteurized soil infested with Pythium isolates and placed in a growth chamber maintained at a constant 16°C and ambient humidity. Plant height, length of the first true leaf, and number of seminal roots were recorded, and roots were digitally scanned to create computer files that were analyzed using WinRhizo software. Pythium isolates caused a significant reduction (P < 0.05) in the number of root tips, root length, and length of the first leaf. Differences in virulence were detected among species and among isolates within species. Isolate Pythium debaryanum 90136 and P. ultimum 90038 were the most virulent and may prove useful in future disease screening assays of Triticum germ plasm.


Carotid artery stenting in patients with high-risk anatomy for carotid endarterectomy.
To report the results of carotid artery stenting (CAS) in patients considered to have high-risk anatomical characteristics for carotid endarterectomy. CAS was performed in 39 carotid arteries of 37 consecutive patients (26 men; mean age 72 +/- 8 years, range 56-88) who met the criteria for high-risk surgical anatomy: previous ipsilateral carotid endarterectomy (20/39, 51.3%), common carotid bifurcation above the mandibular angle (5/39, 12.8%), contralateral carotid artery occlusion (15/39, 38.5%), or previous radiation therapy to the neck (1/39, 2.6%). Palmaz, Integra, or Wallstents were deployed via a percutaneous femoral artery access. Independent neurological evaluation was performed at specified time points, and a dedicated committee adjudicated all clinical events. Procedural success was 100%, with no major in-hospital complications. Neurological events were rare. Only 1 (2.6%) transient ischemic attack occurred prior to discharge; at 30 days, 1 (2.6%) additional minor stroke had been observed, giving a 2.6% cumulative 30-day "death plus any stroke" rate. Over a mean 11 +/- 6-month follow-up, 2 (5.4%) patients died of nonneurological causes, but there were no strokes. CAS is a viable endovascular revascularization technique that can be performed safely and effectively in patients with high-risk anatomy for carotid endarterectomy.


Plasma-assisted deposition of metal fluoride coatings and modeling the extinction coefficient of as-deposited single layers.
We realized metal fluoride coatings with a high packing density and a low extinction coefficient by plasma (ion)-assisted deposition. The densification can be performed by different types of plasma sources, e.g., by a Leybold LION source and a Leybold APSpro, respectively. But the as-deposited coatings show a characteristic absorption behavior, whereas the absorption losses can be reduced in a postdeposition UV treatment step. We show experimental results of the plasma-assisted metal fluorides before and after the UV treatment and present a new model that allows us to describe and calculate the characteristic absorption losses of LaF3, MgF2, and AlF3.


"Relative" adrenal insufficiency in critical illness.
To present a recommended approach to the problem of "relative" adrenal insufficiency (RAI) in the intensive care unit (ICU). We examine historical data that support the traditional concepts of adrenal insufficiency and the idea that the increase in cortisol secretion during stress is needed to survive the stress. The controversial use of treatment with glucocorticoids (GCs) in patients with sepsis and septic shock in the ICU (and thus survival benefit) is also briefly discussed. During the past decade, the concept of RAI as the failure of cortisol secretion to increase in response to stress to sustain the patient through that stress has gained strength. In some studies, it has been suggested that as many as 75% of patients in an ICU setting have RAI. Experimental support for the concept is not possible because there is no clinically useful laboratory measure of GC action. Therefore, diagnosis is generally based on interpretation of the cosyntropin stimulation test. The best clinical judgment should always guide interpretation of any test results, and sharp categorization of patients on the basis of a single cutoff criterion should be avoided. Overall, the concept of RAI has no clinical utility. In these cases, administration of GCs adds cost without benefit and with increased risk.


Comparison of Different Approaches to Surface Functionalization of Biodegradable Polycaprolactone Scaffolds.
Due to their good mechanical stability compared to gelatin, collagen or polyethylene glycol nanofibers and slow degradation rate, biodegradable poly-ε-caprolactone (PCL) nanofibers are promising material as scaffolds for bone and soft-tissue engineering. Here, PCL nanofibers were prepared by the electrospinning method and then subjected to surface functionalization aimed at improving their biocompatibility and bioactivity. For surface modification, two approaches were used: (i) COOH-containing polymer was deposited on the PCL surface using atmospheric pressure plasma copolymerization of CO2 and C2H4, and (ii) PCL nanofibers were coated with multifunctional bioactive nanostructured TiCaPCON film by magnetron sputtering of TiC-CaO-Ti3POx target. To evaluate bone regeneration ability in vitro, the surface-modified PCL nanofibers were immersed in simulated body fluid (SBF, 1×) for 21 days. The results obtained indicate different osteoblastic and epithelial cell response depending on the modification method. The TiCaPCON-coated PCL nanofibers exhibited enhanced adhesion and proliferation of MC3T3-E1 cells, promoted the formation of Ca-based mineralized layer in SBF and, therefore, can be considered as promising material for bone tissue regeneration. The PCL-COOH nanofibers demonstrated improved adhesion and proliferation of IAR-2 cells, which shows their high potential for skin reparation and wound dressing.


Potent PBS/Polysorbate-Soluble Transplatin-Derived Porphyrin-Based Photosensitizers for Photodynamic Therapy.
In this study, we addressed an important drawback of our previously reported tetraplatinated (metallo)porphyrin-based photosensitizers (PSs) for photodynamic therapy (PDT), namely, the poor solubility in aqueous media. We aimed to create tetraplatinated porphyrin-based PSs that are soluble in aqueous media modified with polysorbate (Tween) and do not need to be pre-dissolved in organic solvents. A structural optimization of the previously reported PSs resulted in the synthesis of an extremely potent novel porphyrin-based PS. The novel PS displays effective phototoxicity upon light irradiation against multicellular tumor spheroids and has a phototoxic index (PI) of 6030 in HeLa cells. This PI value is, to the best of our knowledge, the highest value reported for any porphyrin so far.


Increasing stimulus size impairs first- but not second-order motion perception.
As stimulus size increases, the direction of high-contrast moving stimuli becomes increasingly difficult to perceive. This counterintuitive effect, termed spatial suppression, is believed to reflect antagonistic center-surround interactions--mechanisms that play key roles in tasks requiring sensitivity to relative motion. It is unknown, however, whether second-order motion also exhibits spatial suppression. To test this hypothesis, we measured direction discrimination thresholds for first- and second-order stimuli of varying sizes. The results revealed increasing thresholds with increasing size for first-order stimuli but demonstrated no spatial suppression of second-order motion. This selective impairment of first-order motion predicts increasing predominance of second-order cues as stimulus size increases. We confirmed this prediction by utilizing compound stimuli that contain first- and second-order information moving in opposite directions. Specifically, we found that for large stimuli, motion perception becomes increasingly determined by the direction of second-order cues. Overall, our findings show a lack of spatial suppression for second-order stimuli, suggesting that the second-order system may have distinct functional roles, roles that do not require high sensitivity to relative motion.


Expression of rat homeobox gene, rHOX, in developing and adult tissues in mice and regulation of its mRNA expression in osteoblasts by bone morphogenetic protein 2 and parathyroid hormone-related protein.
The rat homeobox gene, rHox, was cloned from a rat osteosarcoma cDNA library. Southwestern and gel mobility shift analyses showed that rHox binds to the promoter regions of collagen (alpha1)I and osteocalcin genes while transient transfection with rHox resulted in repression of their respective promoter activities. In situ hybridization studies showed that rHox mRNA was widely expressed in osteoblasts, chondrocytes, skeletal muscle, skin epidermis, and bronchial and intestinal epithelial cells, as well as cardiac muscle in embryonic and newborn mice. However in 3-month-old mice, rHox mRNA expression was restricted to osteoblasts, megakaryocytes, and myocardium. Bone morphogenetic protein 2, a growth factor that commits mesenchymal progenitor cells to differentiate into osteoblasts, down-regulated rHox mRNA expression by 40-50% in UMR 201, a rat preosteoblast cell line, in a time- and dose-dependent manner. In contrast, PTH-related protein (PTHrP), recently shown to be a negative regulator of chondrocyte differentiation, significantly enhanced rHox mRNA expression in UMR 106-06 osteoblastic cells by 3-fold at 24 h while at the same time down-regulating expression of pro-alpha1(I) collagen mRNA by 60%. Expression of rHox mRNA in calvarial osteoblasts derived from PTHrP -/- mice was approximately 15% of that observed in similar cells obtained from normal mice. In conclusion, current evidence suggests that rHox acts as a negative regulator of osteoblast differentiation. Furthermore, down-regulation of rHox mRNA by bone morphogenetic protein 2 and its up-regulation by PTHrP support a role of the homeodomain protein, rHox, in osteoblast differentiation.


Convection-enhanced delivery of Ls-TPT enables an effective, continuous, low-dose chemotherapy against malignant glioma xenograft model.
Treatment of malignant gliomas represents one of the most formidable challenges in oncology. The combination of surgery, radiation, and chemotherapy yields median survivals of less than one year. Here we demonstrate the use of a minimally invasive surgical technique, convection-enhanced delivery (CED), for local administration of a novel nanoparticle liposome containing topotecan. CED of this liposomal topotecan (Ls-TPT) resulted in extended brain tissue retention (t1/2 = 1.5 days), whereas free topotecan was rapidly cleared (t1/2 = 0.1 days) after CED. The favorable pharmacokinetic profile of extended topotecan release for about seven days, along with biodistribution featuring perivascular accumulation of the nanoparticles, provided, in addition to the known topoisomerase I inhibition, an effective antiangiogenic therapy. In the rat intracranial U87MG tumor model, vascular targeting of Ls-TPT with CED was associated with reductions in laminin expression and vascular density compared to free topotecan or control treatments. A single CED treatment on day 7 showed that free topotecan conferred no survival benefit versus control. However, Ls-TPT produced a significant (P = 0.0002) survival benefit, with six of seven complete cures. Larger U87MG tumors, where CED of Ls-TPT on day 12 resulted in one of six cures, indicated the necessity to cover the entire tumor with the infused therapeutic agent. CED of Ls-TPT was also efficacious in the intracranial U251MG tumor model (P = 0.0005 versus control). We conclude that the combination of a novel nanoparticle Ls-TPT and CED administration was very effective in treating experimental brain tumors.


Gesturing with an injured brain: how gesture helps children with early brain injury learn linguistic constructions.
Children with pre/perinatal unilateral brain lesions (PL) show remarkable plasticity for language development. Is this plasticity characterized by the same developmental trajectory that characterizes typically developing (TD) children, with gesture leading the way into speech? We explored this question, comparing eleven children with PL - matched to thirty TD children on expressive vocabulary - in the second year of life. Children with PL showed similarities to TD children for simple but not complex sentence types. Children with PL produced simple sentences across gesture and speech several months before producing them entirely in speech, exhibiting parallel delays in both gesture + speech and speech-alone. However, unlike TD children, children with PL produced complex sentence types first in speech-alone. Overall, the gesture-speech system appears to be a robust feature of language learning for simple - but not complex - sentence constructions, acting as a harbinger of change in language development even when that language is developing in an injured brain.


A multimedia patient simulation for teaching and assessing endodontic diagnosis.
Teaching and assessing diagnostic skills are difficult due to relatively small numbers of total clinical experiences and a shortage of clinical faculty. Patient simulations could help teach and assess diagnosis by displaying a well-defined diagnostic task, then providing informative feedback and opportunities for repetition and correction of errors. This report describes the development and initial evaluation of SimEndo I, a multimedia patient simulation program that could be used for teaching or assessing endodontic diagnosis. Students interact with a graphical interface that has four pull-down menus and related submenus. In response to student requests, the program presents patient information. Scoring is based on diagnosis of each case by endodontists. Pilot testing with seventy-four junior dental students identified numerous needed improvements to the user interface program. A multi-school field test of the interface program using three patient cases addressed three research questions: 1) How did the field test students evaluate SimEndo I? Overall mean evaluation was 8.1 on a 0 to 10 scale; 2) How many cases are needed to generate a reproducible diagnostic proficiency score for an individual student using the Rimoldi scoring procedure? Mean diagnostic proficiency scores by case ranged from .27 to .40 on a 0 to 1 scale; five cases would produce a score with a 0.80 reliability coefficient; and 3) Did students accurately diagnose each case? Mean correct diagnosis scores by case ranged from .54 to .78 on a 0 to 1 scale. We conclude that multimedia patient simulations offer a promising alternative for teaching and assessing student diagnostic skills.


Specimen volume versus yield in the BACTEC blood culture system.
During a 24-month period, 5,625 blood culture specimens were collected at the Seattle Veterans Administration Medical Center in 20-ml volumes and divided into separate 10-ml aliquots. The two aliquots were processed as duplicate sets (set 1, set 2) by the BACTEC system (Johnston Laboratories, Inc., Towson, Md.). Specimens (5 ml) from each set were inoculated into aerobic (6B) and anaerobic (7C/7D) vials. A total of 434 significantly positive blood cultures were found. In 342 of these positive cultures, yielding 379 isolates (112 members of the family Enterobacteriaceae, 104 staphylococci, 87 streptococci, 27 anaerobes, 20 yeasts, 14 pseudomonads, and 15 miscellaneous organisms), there was adequate specimen volume to fill all four vials. The utilization of set 1 would have resulted only in the failure to detect 65 of 379 (17.2%) significant isolates, 52 of 342 (15.2%) positive cultures, and 20 of 198 (10.1%) bacteremic episodes. There were no significant differences in the recovery of individual species in sets 1 and 2. Although the range of isolates recovered by the aerobic and anaerobic vials of each set differed, the percent yield of total isolates was similar, indicating total isolate yield was predominantly a function of specimen volume. The addition of set 2 most dramatically increased the recovery of Escherichia coli (30%), yeasts (33%), and anaerobes (42%).


Cockroaches probably cleaned up after dinosaurs.
Dinosaurs undoubtedly produced huge quantities of excrements. But who cleaned up after them? Dung beetles and flies with rapid development were rare during most of the Mesozoic. Candidates for these duties are extinct cockroaches (Blattulidae), whose temporal range is associated with herbivorous dinosaurs. An opportunity to test this hypothesis arises from coprolites to some extent extruded from an immature cockroach preserved in the amber of Lebanon, studied using synchrotron X-ray microtomography. 1.06% of their volume is filled by particles of wood with smooth edges, in which size distribution directly supports their external pre-digestion. Because fungal pre-processing can be excluded based on the presence of large particles (combined with small total amount of wood) and absence of damages on wood, the likely source of wood are herbivore feces. Smaller particles were broken down biochemically in the cockroach hind gut, which indicates that the recent lignin-decomposing termite and cockroach endosymbionts might have been transferred to the cockroach gut upon feeding on dinosaur feces.


Calming effects of deep touch pressure in patients with autistic disorder, college students, and animals.
ABSTRACT Many people with autistic disorder have problems with oversensitivity to both touch and sound. The author (an autistic person) developed a device that delivers deep touch pressure to help her learn to tolerate touching and to reduce anxiety and nervousness. The "squeeze machine" applies lateral, inwardly directed pressure to both lateral aspects of a person's entire body, by compressing the user between two foam-padded panels. Clinical observations and several studies suggest that deep touch pressure is therapeutically beneficial for both children with autistic disorder and probably children with attention-deficit hyperactivity disorder. Only minor and occasional adverse effects have been noted. Data are reported that show a similar calming effect in nonreferred college students. A review of the animal literature reveals that animals have similar calming reactions, and also suggests possible additional physiological effects of deep touch pressure. At present, there are increasing anecdotal reports of the clinical value of the squeeze machine, including suggestions that it can be used to reduce required doses of psychostimulant medications. More clinical studies are needed to evaluate the potential role of this seemingly beneficial form of "physiological" stimulation.


Food addiction: an examination of the diagnostic criteria for dependence.
The evidence for food's addictive properties is steadily growing. In addition to clinical and evolutionary plausibility, the possibility of addiction to food is supported by animal model research and increasingly by research with humans. Much as classic drugs of abuse "hijack" the brain, accumulating evidence with food suggests a similar impact, but with weaker effects. Although neurobiological evidence for food addiction is compelling, dependence as conceptualized with respect to alcohol and other drugs of abuse is fundamentally a behavioral disorder. Thus, we review the current state of food addiction research in the context of each of the diagnostic criterion for dependence (ie, tolerance, withdrawal, loss of control) and briefly explore other relevant addiction topics such as expectancies, reinforcement, and incentive salience. There is substantial evidence that some people lose control over their food consumption, suffer from repeated failed attempts to reduce their intake, and are unable to abstain from certain types of food or reduce consumption in the face of negative consequences. Although there is some evidence for other dependence criterion, further research is needed to examine tolerance and withdrawal to high-fat sweets, time spent in obtaining, using, and recovering from excess food consumption and the degree to which important activities are given up due to overconsumption. As science continues forward and both the public and elected leaders become aware that food may trigger an addictive process, this information will likely be used to inform policy. Thus, researchers need to carefully consider the implications of their work and the way in which the results may be interpreted.


Angiomatoid fibrous histiocytoma including cases with pleomorphic features analysed by fluorescence in situ hybridisation.
Angiomatoid fibrous histiocytoma (AFH) is a rare soft-tissue tumour of uncertain differentiation and low metastatic potential. Cytogenetics and/or molecular genetics have revealed that most have a rearrangement of the EWSR1 gene, whereas a FUS gene rearrangement is present in a minority of cases. Although some cases of AFH display striking pleomorphism and mitotic activity, there are no known clinical, morphological or genetic factors that predict metastasis. The authors present clinicopathological features of AFH, including cases showing a pleomorphic histological appearance, and results of fluorescence in situ hybridisation analysis of EWSR1 and FUS rearrangements. Tumour samples from 10 patients were subjected to clinicopathological and immunohistochemical analysis and dual-colour fluorescence in situ hybridisation for EWSR1 and FUS with split-signal probes. All cases showed clinical features (sites: extremities followed by trunk; age: adolescent to young adult), morphology (multinodular proliferation of spindle cells, lymphoid cuffs and pseudovascular spaces) and immunohistochemical results (more than half were positive for CD68, CD99, desmin and epithelial membrane antigen) typical of AFH. There were two local recurrences in each of two patients. Two patients developed distant metastases and died from the disease; tumours of these two patients showed focal proliferation of large pleomorphic cells with hyperchromatic nuclei and high proliferative activity (>10/10 high-power field and Ki-67 labelling index >10%). There were no clinical, histological or immunohistochemical differences between the nine cases with EWSR1 rearrangement and one case with FUS rearrangement. Wide surgical excision and careful follow-up are necessary for patients with AFH in view of its risk of local recurrence and metastasis leading to a fatal outcome.


Vitamin D status in postmenopausal women living at higher latitudes in the UK in relation to bone health, overweight, sunlight exposure and dietary vitamin D.
For 5 months a year the UK has insufficient sunlight for cutaneous synthesis of vitamin D and winter requirements are met from stores made the previous summer. Although there are few natural dietary sources, dietary intake may help maintain vitamin D status. We investigated the relationship between 25-hydroxyvitamin D (25(OH)D), bone health, overweight, sunlight exposure and dietary vitamin D in 3113 women (age 54.8 [SD 2.3] years) living at latitude 57 degrees N between 1998-2000. Serum 25(OH)D was measured by high performance liquid chromatography (HPLC), dietary intakes (food frequency questionnaire, n=2598), sunlight exposure (questionnaire, n=2402) and bone markers were assessed. Bone mineral density (BMD) was measured by dual x-ray absorptiometry in all women at the sampling visit and 6 years before. Seasonal variation in 25(OH)D was not substantial with a peak in the autumn (23.7 [9.9] ng/ml) and a nadir in spring (19.7 [7.6] ng/ml). Daily intake of vitamin D was 4.2 [2.5] mug from food only and 5.8 [4.0] mug including vitamin D from cod liver oil and multivitamins. The latter was associated with 25(OH)D at each season whereas vitamin D simply from food was associated with 25(OH)D in winter and spring only. Sunlight exposure was associated with 25(OH)D in summer and autumn. 25(OH)D was negatively associated with increased bone resorption and bone loss (P<0.05) remaining significant after adjustment for confounders (age, weight, height, menopausal status/HRT use, physical activity and socio-economic status). Using an insufficiency cut-off of <28 ng/ml 25(OH)D, showed lower concentrations of bone resorption markers in the upper category (fDPD/Cr 5.1 [1.7] nmol/mmol compared to 5.3 [2.1] nmol/mmol, P=0.03) and no difference in BMD or bone loss. 25(OH)D was lower (P<0.01) and parathyroid hormone higher (P<0.01) in the top quintile of body mass index. In conclusion, low vitamin D status is associated with greater bone turnover, bone loss and obesity. Diet appears to attenuate the seasonal variation of vitamin D status in early postmenopausal women at northerly latitude where quality of sunlight for production of vitamin D is diminished.


Simultaneous total occlusion of two coronary arteries associated with use of drospirenone-ethinyl estradiol (oral contraceptive).
Although the use of oral contraceptives is associated with an increased risk of venous thromboembolic disease, the risk of myocardial infarction (MI) is unclear. A new, third-generation contraceptive agent, drospirenone-ethinyl estradiol, which contains less estrogen and a new progestogen, drospirenone, in a different combination, has been considered more reliable in terms of risk of MI. However, there have been some cases of MI associated with the use of drospirenone-ethinyl estradiol, despite the protective effects of this oral contraceptive. In this report, a 33-year-old woman who had used drospirenone-ethinyl estradiol for 6 months was admitted with MI and symptoms of cardiogenic shock. Coronary angiography revealed the total occlusion of 2 coronary arteries and so percutaneous coronary intervention was performed. To the best of our knowledge, this is the first case report of simultaneous total occlusion of 2 coronary arteries associated with the use of drospirenone-ethinyl estradiol in the English-language medical literature.


Ca2+ signaling tools acquired from prostasomes are required for progesterone-induced sperm motility.
Progesterone-induced calcium ion (Ca2+) signals in the neck region of sperm play a pivotal role in promoting sperm motility. Here, we show that a long-lasting Ca2+ signal required for sperm motility in response to progesterone depends on their pH-dependent fusion with prostasomes, which are small vesicles secreted by the prostate. We found that prostasome fusion led to the transfer of progesterone receptors, cyclic adenosine diphosphoribose (cADPR)-synthesizing enzymes, ryanodine receptors (RyRs), and other Ca2+ signaling tools from prostasomes to the sperm neck. Progesterone-induced sperm motility relied on cADPR-mediated Ca2+ mobilization through RyR located on acidic Ca2+ stores, followed by Ca2+ entry through store-operated channels. Treatment of prostasome-fused sperm with a cADPR antagonist or fusion with prostasomes in which type 2 RyR was depleted resulted in low fertilization rates, reduced sperm motility, or both. Thus, we conclude that sperm motility depends on the acquisition of Ca2+ signaling tools from prostasomes.


Using guanidine-hydrochloride for fast and efficient protein digestion and single-step affinity-purification mass spectrometry.
Protein digestion is an integral part of the "shotgun" proteomics approach and commonly requires overnight incubation prior to mass spectrometry analysis. Quadruplicate "shotgun" proteomic analysis of whole yeast lysate demonstrated that Guanidine-Hydrochloride (Gnd-HCl) protein digestion can be optimally completed within 30 min with endoprotease Lys-C. No chemical artifacts were introduced when samples were incubated in Gnd-HCl at 95 °C, making Gnd-HCl an appropriate digestion buffer for shotgun proteomics. Current methodologies for investigating protein-protein interactions (PPIs) often require several preparation steps, which prolongs any parallel operation and high-throughput interaction analysis. Gnd-HCl allow the efficient elution and subsequent fast digestion of PPIs to provide a convenient high-throughput methodology for affinity-purification mass spectrometry (AP-MS) experiments. To validate the Gnd-HCl approach, label-free PPI analysis of several GFP-tagged yeast deubiquitinating enzymes was performed. The identification of known interaction partners demonstrates the utility of the optimized Gnd-HCl protocol that is also scalable to the 96 well-plate format.


The Brain Anatomy of the Brown Bear (Carnivora, Ursus arctos L., 1758) Compared to That of Other Carnivorans: A Cross-Sectional Study Using MRI.
In this study, we aimed to provide a neuroanatomy atlas derived from cross-sectional and magnetic resonance imaging (MRI) of the encephalon of the brown bear (Ursus arctos). A postmortem brain analysis using magnetic resonance imaging (MRI - 1,5T; a high-resolution submillimeter three-dimensional T1-3D FFE) and cross-sectional macroscopic anatomy methods revealed major embryological and anatomical subdivisions of the encephalon, including the ventricular system. Most of the internal structures were comparably identifiable in both methods. The tractus olfactorius medialis, corpus subthalamicum, brachium colliculi rostralis, fasciculus longitudinalis medialis, nuclei vestibulares, velum medullare rostrale, nucleus fastigii, fasciculi cuneatus et gracilis were identified entirely by cross-sectional macroscopic analysis. However, the glandula pinealis, lemniscus lateralis and nuclei rhaphe were visualized only with MRI. Gross neuroanatomic analysis provided information about sulci and gyri of the cerebral hemispheres, components of the vermis and cerebellar hemispheres, and relative size and morphology of constituents of the rhinencephalon and cerebellum constituents. Similarities and discrepancies in identification of structures provided by both methods, as well as hallmarks of the structures facilitating identification using these methods are discussed. Finally, we compare the brown bear encephalon with other carnivores and discuss most of the identified structures compared to those of the domestic dog, the domestic cat, Ursidae and Mustelidae families and Pinnipedia clade.


Effect of Low-Level Laser Therapy (LLLT) in Pulmonary Inflammation in Asthma Induced by House Dust Mite (HDM): Dosimetry Study.
Asthma is characterized by chronic inflammation in the airways. Several models have been proposed for the discovery of new therapies. Low-Level Laser Therapy (LLLT) is relatively new and effective, very low cost, with no side effects. However, there is still no consensus on the optimal dose to be used. In this sense, the objective of the present study was to evaluate the best dose in an experimental model of asthma induced by House Dust Mite (HDM). Balb/c mice received administration of 100 ug/animal HDM and LLLT applications (diode laser: 660 nm, 100 mW and four different energies 1J, 3J, 5J, and 7.5J) for 16 days. After 24 hours, we studied inflammatory, functional, and structural parameters. The results showed that LBI was able to modulate the pulmonary inflammation observed by reducing the number of cells in Bronchoalveolar Lavage Fluid (BALF) as well as reducing the percentage of neutrophils, eosinophils and T lymphocytes. On the other hand, LLLT increased the level of IL-10 and reduced levels of IL-4, IL-5 and IL-13 in BALF. LLLT was able to reduce the production of mucus, peribronchial eosinophils, collagen deposition, bronchoconstriction index, and bronchial and muscular thickening in the airways. We concluded that the use of LLLT in the treatment of chronic inflammation of the airways attenuated the inflammatory process and functional and structural parameters. We emphasize, in general, that the 1J and 3J laser presented better results. Thus, photobiomodulation may be considered a promising tool for the treatment of chronic pulmonary allergic inflammation observed in asthma.


Measles virus-induced suppression of immune responses.
Measles is an important cause of child mortality that has a seemingly paradoxical interaction with the immune system. In most individuals, the immune response is successful in eventually clearing measles virus (MV) infection and in establishing life-long immunity. However, infection is also associated with persistence of viral RNA and several weeks of immune suppression, including loss of delayed type hypersensitivity responses and increased susceptibility to secondary infections. The initial T-cell response includes CD8+ and T-helper 1 CD4+ T cells important for control of infectious virus. As viral RNA persists, there is a shift to a T-helper 2 CD4+ T-cell response that likely promotes B-cell maturation and durable antibody responses but may suppress macrophage activation and T-helper 1 responses to new infections. Suppression of mitogen-induced lymphocyte proliferation can be induced by lymphocyte infection with MV or by lymphocyte exposure to a complex of the hemagglutinin and fusion surface glycoproteins without infection. Dendritic cells (DCs) are susceptible to infection and can transmit infection to lymphocytes. MV-infected DCs are unable to stimulate a mixed lymphocyte reaction and can induce lymphocyte unresponsiveness through expression of MV glycoproteins. Thus, multiple factors may contribute both to measles-induced immune suppression and to the establishment of durable protective immunity.


Changes in estradiol predict within-women shifts in attraction to facial cues of men's testosterone.
Many studies have demonstrated that women express stronger attraction to androgen-related traits when tested near ovulation than when tested at other times in the cycle. Much less research, however, has directly addressed which hormonal or other physiological signals may regulate these temporal shifts in women's attractiveness judgments. In the present study, we measured women's preferences for facial cues of men's testosterone concentrations on two occasions spaced two weeks apart, while also measuring women's salivary estradiol and testosterone concentrations at each testing session. Changes in women's estradiol concentrations across sessions positively predicted changes in their preferences for facial cues of high testosterone; there was no such effect for changes in women's testosterone concentrations. For the subset of women who had a testing session fall within the estimated fertile window, preferences for high testosterone faces were stronger in the fertile window session, and change in estradiol from outside to inside the fertile window positively predicted the magnitude of the ovulatory preference shift. These patterns were not replicated when testing preferences for faces that were rated as high in masculinity, suggesting that facial cues of high testosterone can be distinguished from the cues used to subjectively judge facial masculinity. Our findings suggest that women's estradiol promotes attraction to androgen-dependent cues in men (similar to its effects in females of various nonhuman species), and support a role for this hormone as a physiological regulator of cycle phase shifts in mating psychology.


A dynamic mode of mitotic bookmarking by transcription factors.
During mitosis, transcription is shut off, chromatin condenses, and most transcription factors (TFs) are reported to be excluded from chromosomes. How do daughter cells re-establish the original transcription program? Recent discoveries that a select set of TFs remain bound on mitotic chromosomes suggest a potential mechanism for maintaining transcriptional programs through the cell cycle termed mitotic bookmarking. Here we report instead that many TFs remain associated with chromosomes in mouse embryonic stem cells, and that the exclusion previously described is largely a fixation artifact. In particular, most TFs we tested are significantly enriched on mitotic chromosomes. Studies with Sox2 reveal that this mitotic interaction is more dynamic than in interphase and is facilitated by both DNA binding and nuclear import. Furthermore, this dynamic mode results from lack of transcriptional activation rather than decreased accessibility of underlying DNA sequences in mitosis. The nature of the cross-linking artifact prompts careful re-examination of the role of TFs in mitotic bookmarking.


The Effects of Changing Attention and Context in an Awake Offline Processing Period on Visual Long-Term Memory.
There is accumulating evidence that sleep as well as awake offline processing is important for the transformation of new experiences into long-term memory (LTM). Yet much remains to be understood about how various cognitive factors influence the efficiency of awake offline processing. In the present study we investigated how changes in attention and context in the immediate period after exposure to new visual information influences LTM consolidation. After presentation of multiple naturalistic scenes within a working memory paradigm, recognition was assessed 30 min and 24 h later in three groups of subjects. One group of subjects engaged in a focused attention task [the Revised Attentional Network Task (R-ANT)] in the 30 min after exposure to the scenes. Another group of subjects remained in the testing room during the 30 min after scene exposure and engaged in no goal- or task-directed activities. A third group of subjects left the testing room and returned 30 min later. A signal detection analysis revealed no significant differences among the three groups in hits, false alarms, or sensitivity on the 30-min recognition task. At the 24-h recognition test, the group that performed the R-ANT made significantly fewer hits compared to the group that left the testing room and did not perform the attention ask. The group that performed the R-ANT and the group that remained in the testing room during the 30-min post-exposure interval made significantly fewer false alarms on the 24-h recognition test compared to the group that left the testing room. The group that stayed in the testing room and engaged in no goal- or task-directed activities exhibited significantly higher sensitivity (d') compared to the group that left the testing room and the group that performed the R-ANT task. Staying in the same context after exposure to new information and resting quietly with minimal engagement of attention results in the best ability to distinguish old from novel visual stimuli after 24 h. These findings suggest that changes in attentional demands and context during an immediate post-exposure offline processing interval modulate visual memory consolidation in a subtle but significant manner.


Coexistence of cooperation and defection in public goods games.
The production of public goods by the contribution of individual volunteers is a social dilemma because an individual that does not volunteer can benefit from the public good produced by the contributions of others. Therefore it is generally believed that public goods can be produced only in the presence of repeated interactions (which allow reciprocation, reputation effects and punishment) or relatedness (kin selection). Cooperation, however, often occurs in the absence of iterations and relatedness. We show that when the production of a public good is a Volunteer's Dilemma, in which a fixed number of cooperators is necessary to produce the public good, cooperators and defectors persist in a mixed equilibrium, without iterations and without relatedness. This mixed equilibrium is absent in the N-person Prisoner's Dilemma, in which the public good is a linear function of the individual contributions. We also show that the Prisoner's Dilemma and the Volunteer's Dilemma are the two opposite extremes of a general public goods game, and that all intermediate cases can have a mixed equilibrium like the Volunteer's Dilemma. The coexistence of cooperators and defectors, therefore, is a typical outcome of most social dilemmas, which requires neither relatedness nor iterations.


Proteomic response to acupuncture treatment in spontaneously hypertensive rats.
Previous animal and clinical studies have shown that acupuncture is an effective alternative treatment in the management of hypertension, but the mechanism is unclear. This study investigated the proteomic response in the nervous system to treatment at the Taichong (LR3) acupoint in spontaneously hypertensive rats (SHRs). Unanesthetized rats were subject to 5-min daily acupuncture treatment for 7 days. Blood pressure was monitored over 7 days. After euthanasia on the 7(th) day, rat medullas were dissected, homogenized, and subject to 2D gel electrophoresis and MALDI-TOF analysis. The results indicate that blood pressure stabilized after the 5th day of acupuncture, and compared with non-acupoint treatment, Taichong-acupunctured rat's systolic pressure was reduced significantly (P<0.01), though not enough to bring blood pressure down to normal levels. The different treatment groups also showed differential protein expression: the 2D images revealed 571 ± 15 proteins in normal SD rats' medulla, 576 ± 31 proteins in SHR's medulla, 597 ± 44 proteins in medulla of SHR after acupuncturing Taichong, and 616 ± 18 proteins in medulla of SHR after acupuncturing non-acupoint. In the medulla of Taichong group, compared with non-acupoint group, seven proteins were down-regulated: heat shock protein-90, synapsin-1, pyruvate kinase isozyme, NAD-dependent deacetylase sirtuin-2, protein kinase C inhibitor protein 1, ubiquitin hydrolase isozyme L1, and myelin basic protein. Six proteins were up-regulated: glutamate dehydrogenase 1, aldehyde dehydrogenase 2, glutathione S-transferase M5, Rho GDP dissociation inhibitor 1, DJ-1 protein and superoxide dismutase. The altered expression of several proteins by acupuncture has been confirmed by ELISA, Western blot and qRT-PCR assays. The results indicate an increase in antioxidant enzymes in the medulla of the SHRs subject to acupuncture, which may provide partial explanation for the antihypertensive effect of acupuncture. Further studies are warranted to investigate the role of oxidative stress modulation by acupuncture in the treatment of hypertension.


The clinical spectrum of leukocyte adhesion deficiency (LAD) III due to defective CalDAG-GEF1.
Leukocyte adhesion deficiency (LAD) type III is a rare syndrome characterized by severe recurrent infections, leukocytosis, and increased bleeding tendency. All integrins are normally expressed yet a defect in their activation leads to the observed clinical manifestations. Less than 20 patients have been reported world wide and the primary genetic defect was identified in some of them. Here we describe the clinical features of patients in whom a mutation in the calcium and diacylglycerol-regulated guanine nucleotide exchange factor 1 (CalDAG GEF1) was found and compare them to other cases of LAD III and to animal models harboring a mutation in the CalDAG GEF1 gene. The hallmarks of the syndrome are recurrent infections accompanied by severe bleeding episodes distinguished by osteopetrosis like bone abnormalities and neurodevelopmental defects.


Attenuated familial adenomatous polyposis: results from an international collaborative study.
The study aimed to describe genetical and clinical features of attenuated familial adenomatous polyposis (AFAP) and to propose clinical criteria and guidelines for treatment and surveillance. A questionnaire study was carried out of polyposis registries with data on patients with presumed AFAP, defined as having ≤ 100 colorectal adenomas at age ≥ 25. One hundred and ninety-six patients were included. The median number of adenomas was 25 (0-100) with a uniform distribution of colorectal adenomas and carcinomas (CRC). Age at CRC diagnosis was delayed by 15 years compared with classic FAP. Eighty-two patients had a colectomy and an ileorectal anastomosis and 5/82 (6%) had a secondary proctectomy. The location of the mutation in the APC gene was known in 69/171 (40%) tested patients. Only 15/29 (52%) of mutations in APC were found in parts of the gene usually associated with AFAP (the 5' end, exon 9 and 3' end). A subset of FAP patients with a milder phenotype does exist and treatment and surveillance had to be modified accordingly. The mutation detection rate is lower than in classic FAP and mutations in AFAP patients are located throughout the APC gene. We propose the following clinical diagnostic criteria for AFAP: a dominant mode of inheritance of colorectal adenomatosis and <100 colorectal adenomas at age 25 or older. Colonoscopy had to be preferred to sigmoidoscopy and surveillance had to be life-long. In the majority of patients, prophylactic colectomy and ileorectal anastomosis are recommended at the age of 20-25 years.


Zinc-mediated α-regioselective Barbier-type cinnamylation reactions of aldehydes, ketones and esters.
We report a simple, efficient, and general method for the zinc-mediated regioselective cinnamylation of aldehydes and ketones under Barbier-type conditions in a one-pot synthesis affording the corresponding α-cinnamylated alcohols in moderate to excellent yields. Compared to the literature procedures, this approach is operationally simple, uses simple reactants, and provides direct access to linear α-cinnamylated alcohols with excellent regioselectivity. Experimental results suggest that the reactions proceed through the radical pathway. In addition, the reaction was found to be scalable to the gram-scale and the one-pot protocol is also applicable to less reactive esters leading to bishomoallylic alcohols which were valuable intermediates for desymmetrizing intramolecular Heck cyclization, allowing for the elaboration to functionalized building blocks.


Protecting the frontline: designing an infection prevention platform for preventing emerging respiratory viral illnesses in healthcare personnel.
Healthcare personnel often find themselves on the frontlines of any epidemic, and may be at particularly high risk of acquiring respiratory viral illnesses when compared to the general population. Many aspects dictate how respiratory viruses spread both inside the hospital and out: Elements to consider include the specific type of virus being targeted for prevention, as well as environmental conditions and host factors, such as age and immune status. Due to the diverse nature of these agents, multiple modes of transmission, including contact, droplet, aerosol, and transocular, must be considered when designing an effective infection prevention program. In this review, we examine the data behind current theories of respiratory virus transmission and key elements of any respiratory illness prevention program. We also highlight other influences that may come into play, such as the cost-effectiveness of choosing one respiratory protection strategy over another.


Multicolour single molecule emission and excitation spectroscopy reveals extensive spectral shifts.
We explore the distribution and shape of single molecule spectra at room temperature, when embedded in a polymer host. Multicolour excitation and emission spectroscopy is implemented to capture the full inhomogeneous distribution. We observe dramatic spectral changes in a distribution of single quaterrylene diimide (QDI) molecules isolated in a PMMA matrix. The molecules are strongly blue shifted with respect to the ensemble absorption maximum and spread over a staggering 200 nm range. Despite these strong shifts, the shape of the emission spectra does not differ much between individual molecules. We demonstrate that a considerable number of molecules may be invisible in single molecule experiments, as they typically rely on only a single excitation wavelength, which predetermines which subensemble is probed in the experiment. Lastly, we make a first step towards single molecule excitation spectroscopy under ambient conditions, which allows us to determine the spectral range at which individual molecules absorb light most efficiently. We show how single molecule emission and excitation spectroscopies can complement each other and a combination of both techniques can help in understanding the origin of underlaying spectral properties of individual molecules.


p300 cooperates with c-Jun and PARP-1 at the p300 binding site to activate RhoB transcription in NSC126188-mediated apoptosis.
The anti-cancer agent NSC126188 induces apoptosis of stomach carcinoma NUGC-3 cells by inducing RhoB expression. Here, we present that the p300 binding site in the RhoB promoter is crucial for the binding of p300 and its partner transcription factors to activate RhoB transcription in NSC126188-mediated apoptosis. NSC126188 increased expression of p300 and c-Jun. Conversely, knockdown of p300 decreased RhoB expression in the presence of NSC126188. We found that poly(ADP-ribose) polymerase-1 (PARP-1) was associated with the p300 binding site and that PARP-1 knockdown inhibited NSC126188-mediated RhoB expression. In the cells treated with NSC126188, p300, PARP-1, and c-Jun interacted and bound the p300 binding site. Furthermore, chromatin immunoprecipitation (ChIP) analysis revealed strong p300 binding and weak c-Jun binding at the p300 binding site of RhoB promoter in cells treated with NSC126188. We also demonstrated that c-Jun played a crucial role in p300 binding. However, PARP-1 did not directly bind the p300 binding site, suggesting a bridging role between p300 and c-Jun. Electrophoretic mobility shift assays demonstrated a complex comprising p300/c-Jun/PARP-1 that bound wild type, but not a mutated, p300 binding site. In addition, overexpression of p300, PARP-1, or c-Jun dramatically enhanced RhoB promoter activity when it contained the wild type sequence but not mutated sequences, indicating the crucial role of the p300 binding site in NSC126188-induced transcription of RhoB. Taken together, these data suggest that p300 is recruited and cooperates with c-Jun and PARP-1 at the p300 binding site to activate RhoB transcription during NSC126188-mediated apoptosis.