Subpopulation of Macrophage-Like Plasmatocytes Attenuates Systemic Growth via JAK/STAT in the Drosophila Fat Body.
Drosophila hemocytes, like those of mammals, are given rise from two distinctive phases during both the embryonic and larval hematopoiesis. Embryonically derived hemocytes, mostly composed of macrophage-like plasmatocytes, are largely identified by genetic markers. However, the cellular diversity and distinct functions of possible subpopulations within plasmatocytes have not been explored in Drosophila larvae. Here, we show that larval plasmatocytes exhibit differential expressions of Hemolectin (Hml) and Peroxidasin (Pxn) during development. Moreover, removal of plasmatocytes by overexpressing pro-apoptotic genes, hid and reaper in Hml-positive plasmatocytes, feeding high sucrose diet, or wasp infestation results in increased circulating hemocytes that are Hml-negative. Interestingly these Hml-negative plasmatocytes retain Pxn expression, and animals expressing Hml-negative and Pxn-positive subtype largely attenuate growth and abrogate metabolism. Furthermore, elevated levels of a cytokine, unpaired 3, are detected when Hml-positive hemocytes are ablated, which in turn activates JAK/STAT activity in several tissues including the fat body. Finally, we observed that insulin signaling is inhibited in this background, which can be recovered by concurrent loss of upd3. Overall, this study highlights heterogeneity in Drosophila plasmatocytes and a functional plasticity of each subtype, which reaffirms extension of their role beyond immunity into metabolic regulation for cooperatively maintaining internal homeostatic balance.


Effectiveness of the Open Screening Programs in Recruiting Subjects to Prodromal and Mild Alzheimer's Disease Clinical Trials.
Due to the lack of scientific data comparing the success and cost-effectiveness of trial recruiting strategies, the main goal of this paper is to present our results and experiences in recruiting participants to prodromal and mild AD clinical trials from an open-access screening program. The screening procedure includes the interview, and combined tests administration conducted by experienced neuropsychologist: Mini-Mental State Examination (MMSE) and Auditory-Verbal Learning Test (AVLT). The clinical evaluation was based on test scores, patient and/or caregiver interview, and the health questionnaire. The open-access screening program was conducted in Wroclaw Alzheimer's Center for 18 months (2018-2019). We invited individuals age 50 or older with the caregivers. The total number of subjects was 730 (N=730). Due to our research, the detection rates in the screened population were 0,7% for severe dementia, 4,1% for moderate dementia, 18,6% for mild dementia, and 28,9% for mild cognitive impairment (MCI). From 347 individuals classified in our open-access screening programs as MCI or mild dementia patients, as many as 248 patients were screened in Alzheimer's disease clinical trials, which is 71,47%. Moreover, 63 from 347 individuals selected from our program as MCI or mild dementia patients were randomized into the clinical trials, which is 18,16%. Furthermore, 63 from total 730 (8,6%) patients were randomized in clinical trials. Open-access screening programs can improve detection of MCI and dementia in society, help to distinguish demented from non-demented elderly, and improve recruitment of prodromal AD patients who would probably not have come to the memory clinic otherwise.


Origins of life: a problem for physics, a key issues review.
The origins of life stands among the great open scientific questions of our time. While a number of proposals exist for possible starting points in the pathway from non-living to living matter, these have so far not achieved states of complexity that are anywhere near that of even the simplest living systems. A key challenge is identifying the properties of living matter that might distinguish living and non-living physical systems such that we might build new life in the lab. This review is geared towards covering major viewpoints on the origin of life for those new to the origin of life field, with a forward look towards considering what it might take for a physical theory that universally explains the phenomenon of life to arise from the seemingly disconnected array of ideas proposed thus far. The hope is that a theory akin to our other theories in fundamental physics might one day emerge to explain the phenomenon of life, and in turn finally permit solving its origins.


The role of CXCL12 in the organ-specific process of artery formation.
CXC chemokine ligand 12 (CXCL12; stromal cell-derived factor-1 [SDF-1]/pre-B-cell growth-stimulating factor [PBSF]) and its receptor CXCR4 are essential for vascularization in the gastrointestinal tract as well as B lymphopoiesis and colonization of bone marrow by hematopoietic cells. However, the mechanism by which CXCL12/CXCR4 functions in blood vessel formation remains elusive. Here, we have found a novel mode of organ vascularization and determined the roles of CXCL12 in these processes. In the developing small intestine, many short interconnecting vessels form between larger superior mesenteric artery (SMA) and the neighboring primary capillary plexus surrounding the primitive gut, and they elongate and become the arteries supplying the small intestine. Mice lacking CXCL12 or CXCR4 lack the interconnecting vessels but have normal venous networks. The mutants lack filopodial extension and intussusception from endothelial cells of SMAs seen in wild-type embryos. CXCR4 is specifically expressed in arteries in the developing mesenteries and its expression is severely reduced in CXCL12-/- embryos. Mice in which CXCR4 is specifically deleted in the endothelium reveal vascular defects identical to those observed in the conventional CXCR4-/- embryos. Together, CXCL12 acts on arterial endothelial cells of SMA to up-regulate CXCR4 and mediate the connection between the larger artery and neighboring capillary plexus in an organ-specific manner.


Dielectric ordering of water molecules arranged in a dipolar lattice.
Intermolecular hydrogen bonds impede long-range (anti-)ferroelectric order of water. We confine H2O molecules in nanosized cages formed by ions of a dielectric crystal. Arranging them in channels at a distance of ~5 Å with an interchannel separation of ~10 Å prevents the formation of hydrogen networks while electric dipole-dipole interactions remain effective. Here, we present measurements of the temperature-dependent dielectric permittivity, pyrocurrent, electric polarization and specific heat that indicate an order-disorder ferroelectric phase transition at T0 ≈ 3 K in the water dipolar lattice. Ab initio molecular dynamics and classical Monte Carlo simulations reveal that at low temperatures the water molecules form ferroelectric domains in the ab-plane that order antiferroelectrically along the channel direction. This way we achieve the long-standing goal of arranging water molecules in polar order. This is not only of high relevance in various natural systems but might open an avenue towards future applications in biocompatible nanoelectronics.


Drinking water arsenic exposure and blood pressure in healthy women of reproductive age in Inner Mongolia, China.
The extremely high exposure levels evaluated in prior investigations relating elevated levels of drinking water arsenic and hypertension prevalence make extrapolation to potential vascular effects at lower exposure levels very difficult. A cross-sectional study was conducted on 8790 women who had recently been pregnant in an area of Inner Mongolia, China known to have a gradient of drinking water arsenic exposure. This study observed increased systolic blood pressure levels with increasing drinking water arsenic, at lower exposure levels than previously reported in the literature. As compared to the referent category (below limit of detection to 20 microg of As/L), the overall population mean systolic blood pressure rose 1.29 mm Hg (95% CI 0.82, 1.75), 1.28 mm Hg (95% CI 0.49, 2.07), and 2.22 mm Hg (95% CI 1.46, 2.97) as drinking water arsenic concentration increased from 21 to 50, 51 to 100, and >100 microg of As/L, respectively. Controlling for age and body weight (n=3260), the population mean systolic blood pressure rose 1.88 mm Hg (95% CI 1.03, 2.73), 3.90 mm Hg (95% CI 2.52, 5.29), and 6.83 mm Hg (95% CI 5.39, 8.27) as drinking water arsenic concentration increased, respectively. For diastolic blood pressure effect, while statistically significant, was not as pronounced as systolic blood pressure. Mean diastolic blood pressure rose 0.78 mm Hg (95% CI 0.39, 1.16), 1.57 mm Hg (95% CI 0.91, 2.22) and 1.32 mm Hg (95% CI 0.70, 1.95), respectively, for the overall population and rose 2.11 mm Hg (95% CI 1.38, 2.84), 2.74 mm Hg (95% CI 1.55, 3.93), and 3.08 mm Hg (95% CI 1.84, 4.31), respectively, for the adjusted population (n=3260) at drinking water arsenic concentrations of 21 to 50, 51 to 100, and >100 microg of As/L. If our study results are confirmed in other populations, the potential burden of cardiovascular disease attributable to drinking water arsenic is significant.


When More Is Not Better: Three Common Mistakes in Health Messaging Interventions.
Health messaging interventions frequently make three well-intentioned but mistaken choices in their communications strategies. To increase their persuasiveness, these messages frequently call attention to the greatest possible numbers of people engaging in undesirable behavior, victims of this behavior, and reasons why one should change the behavior. We raise recent research suggesting how and why the intuitively attractive more-is-better heuristic can be unproductive, and suggest ways to overcome these pitfalls.


An organic self-regulating microfluidic system.
In this paper we present an organic feedback scheme that merges microfluidics and responsive materials to address several limitations of current microfluidic systems. By using in situ fabrication and by taking advantage of microscale phenomena (e.g., laminar flow, short diffusion times), we have demonstrated feedback control of the output pH in a completely organic system. The system autonomously regulates an output stream at pH 7 under a range of input flow conditions. A single responsive hydrogel component performs the functionality of traditional feedback system components. Vertically stacked laminar flow is used to improve the time response of the hydrogel actuator. A star shaped orifice is utilized to improve the flow characteristics of the membrane/orifice valve. By changing the chemistry of the hydrogel component, the system can be altered to regulate flow based on hydrogels sensitive to temperature, light, biological/molecular, and others.


Mentoring the Mentors: Implementation and Evaluation of Four Fogarty-Sponsored Mentoring Training Workshops in Low-and Middle-Income Countries.
A growing body of evidence highlights the importance of competent mentoring in academic research. We describe the development, implementation, and evaluation of four regional 2-day intensive workshops to train mid- and senior-level investigators conducting public health, clinical, and basic science research across multiple academic institutions in low- and middle-income countries (LMICs) on tools and techniques of effective mentoring. Sponsored by the Fogarty International Center, workshops included didactic presentations, interactive discussions, and small-group problem-based learning and were conducted in Lima, Peru; Mombasa, Kenya; Bangalore, India; and Johannesburg, South Africa, from 2013 to 2016. Mid- or senior-level faculty from multiple academic institutions within each region applied and were selected. Thirty faculty from 12 South America-based institutions, 29 faculty from eight East Africa-based institutions, 37 faculty from 14 South Asia-based institutions, and 36 faculty from 13 Africa-based institutions participated, with diverse representation across disciplines, gender, and academic rank. Discussions and evaluations revealed important comparisons and contrasts in the practice of mentoring, and specific barriers and facilitators to mentoring within each cultural and regional context. Specific regional issues related to hierarchy, the post-colonial legacy, and diversity arose as challenges to mentoring in different parts of the world. Common barriers included a lack of a culture of mentoring, time constraints, lack of formal training, and a lack of recognition for mentoring. These workshops provided valuable training, were among the first of their kind, were well-attended, rated highly, and provided concepts and a structure for the development and strengthening of formal mentoring programs across LMIC institutions.


Stillbirth Collaborative Research Network: design, methods and recruitment experience.
The Stillbirth Collaborative Research Network (SCRN) has conducted a multisite, population-based, case-control study, with prospective enrollment of stillbirths and livebirths at the time of delivery. This paper describes the general design, methods and recruitment experience. The SCRN attempted to enroll all stillbirths and a representative sample of livebirths occurring to residents of pre-defined geographical catchment areas delivering at 59 hospitals associated with five clinical sites. Livebirths <32 weeks gestation and women of African descent were oversampled. The recruitment hospitals were chosen to ensure access to at least 90% of all stillbirths and livebirths to residents of the catchment areas. Participants underwent a standardised protocol including maternal interview, medical record abstraction, placental pathology, biospecimen testing and, in stillbirths, post-mortem examination. Recruitment began in March 2006 and was completed in September 2008 with 663 women with a stillbirth and 1932 women with a livebirth enrolled, representing 69% and 63%, respectively, of the women identified. Additional surveillance for stillbirths continued until June 2009 and a follow-up of the case-control study participants was completed in December 2009. Among consenting women, there were high consent rates for the various study components. For the women with stillbirths, 95% agreed to a maternal interview, chart abstraction and a placental pathological examination; 91% of the women with a livebirth agreed to all of these components. Additionally, 84% of the women with stillbirths agreed to a fetal post-mortem examination. This comprehensive study is poised to systematically study a wide range of potential causes of, and risk factors for, stillbirths and to better understand the scope and incidence of the problem.


Qualitative review of intensive care unit quality indicators.
The purpose of this study was to (1) conduct a systematic review of the literature to identify interventions that improve patient outcomes in the intensive care unit (ICU); (2) evaluate potential measures of quality based on the impact, feasibility, variability, and the strength of evidence to support each measure and to categorize these measures as outcome, process, access, or complication measures; and (3) select a list of candidate quality measures that can be broadly applied to improve ICU care. We identified and independently reviewed all studies in Medline (1965-2000) and The Cochrane Library (Issue 3, 2001) that met the following criteria: design: observational studies, experimental trials, or systematic reviews; population: critically ill adults; and intervention: process or structure measure that was associated with improved patient outcomes: morbidity, mortality, complications, errors, costs, length of stay (LOS), and patient reported outcomes. Studies were grouped into categories by the type of outcome that was improved by the intervention. Potential quality measures were evaluated for: impact on morbidity, mortality, and costs; feasibility of the measure; and variability in the measure. We evaluated the strength of evidence for each intervention used to improve outcomes and using the Delphi method, assigned an over-all recommendation for each quality measure. A total of 3,014 citations were identified. Sixty-six studies that met selection criteria reported on a variety of interventions that were associated with improved patient outcomes. We identified 6 outcome measures: ICU mortality rate, ICU LOS greater than 7 days, average ICU LOS, average days on mechanical ventilation, suboptimal management of pain, and patient/family satisfaction; 6 process measures: effective assessment of pain, appropriate use of blood transfusions, prevention of ventilator-associated pneumonia, appropriate sedation, appropriate peptic ulcer disease prophylaxis, and appropriate deep venous thrombosis prophylaxis; 4 access measures: rate of delayed admissions, rate of delayed discharges, cancelled surgical cases, and emergency department by-pass hours; and 3 complication measures: rate of unplanned ICU readmission, rate of catheter-related blood stream infections, and rate of resistant infections. Further work is needed to create operational definitions and to pilot test the selected measures. The value of these measures will be determined by our ability to evaluate our current performance and implement interventions designed to improve the quality of ICU care.


Yellow fever in the diagnostics laboratory.
Yellow fever (YF) remains a public health issue in endemic areas despite the availability of a safe and effective vaccine. In 2015-2016, urban outbreaks of YF were declared in Angola and the Democratic Republic of Congo, and a sylvatic outbreak has been ongoing in Brazil since December 2016. Of great concern is the risk of urban transmission cycles taking hold in Brazil and the possible spread to countries with susceptible populations and competent vectors. Vaccination remains the cornerstone of an outbreak response, but a low vaccine stockpile has forced a sparing-dose strategy, which has thus far been implemented in affected African countries and now in Brazil. Accurate laboratory confirmation of cases is critical for efficient outbreak control. A dearth of validated commercial assays for YF, however, and the shortcomings of serological methods make it challenging to implement YF diagnostics outside of reference laboratories. We examine the advantages and drawbacks of existing assays to identify the barriers to timely and efficient laboratory diagnosis. We stress the need to develop new diagnostic tools to meet current challenges in the fight against YF.


Racial and Ethnic Diversity in Grounded Theory Research.
National initiatives in the United States call for health research that addresses racial/ethnic disparities. Although grounded theory (GT) research has the potential to contribute much to the understanding of the health experiences of people of color, the extent to which it has contributed to health disparities research is unclear. In this article we describe a project in which we reviewed 44 GT studies published in Qualitative Health Research within the last five years. Using a framework proposed by Green, Creswell, Shope, and Clark (2007), we categorized the studies at one of four levels based on the status and significance afforded racial/ethnic diversity. Our results indicate that racial/ethnic diversity played a primary role in five studies, a complementary role in one study, a peripheral role in five studies, and an absent role in 33 studies. We suggest that GT research could contribute more to health disparities research if techniques were developed to better analyze the influence of race/ethnicity on health-related phenomena.


Meiotic activation of rat pachytene spermatocytes with okadaic acid: the behaviour of synaptonemal complex components SYN1/SCP1 and COR1/SCP3.
The phosphatase inhibitor okadaic acid accelerates meiotic events in rodent germ cells in culture. Isolated pachytene spermatocytes treated with okadaic acid proceed to a metaphase I arrest in a few hours as opposed to the similar process in vivo, which requires several days. Leptotene/zygotene spermatocytes cannot be activated in this way, suggesting that okadaic acid enables cells to bypass a sensor of the meiotic progression, which is pachytene specific. We monitored the chromosome behaviour accompanying the transition to metaphase I in rat spermatocytes with antibodies against COR1/SCP3, a component of the meiotic chromosome cores, and against the synaptic protein, SYN1/SCP1. Okadaic acid induced a rapid synaptonemal complex dissolution and bivalent separation, followed by chromosome condensation and chiasmata formation, similar to the succession of events in untreated cells. The similarity between meiosis I induced with okadaic acid and the meiosis I events in vivo extends to the dissolution of the nuclear membrane and the disappearance of the microtubule network at the onset of metaphase I. This cell culture system provides a model for the in vivo transition from pachytene to metaphase I and therefore can be used in the study of this transition at the molecular level. The effect of okadaic acid is most likely mediated by the activation of tyrosine kinases, as addition of genistein, a general tyrosine kinase inhibitor, completely abolishes the observed effect of okadaic acid on chromosome metabolism. The okadaic acid-induced progression to the metaphase I arrest is not affected by the inhibition of protein synthesis. However, pachytene spermatocytes incubated in the presence of protein synthesis inhibitors for 6 hours show loss of synapsis which is abnormal in that it is not accompanied by chiasmata formation. The two meiosis-specific proteins, SYN1/SCP1 and COR1/SCP3, are efficiently phosphorylated in vitro by extracts from isolated pachytene cells. Extracts from cells that have reached metaphase I upon okadaic acid treatment, with concomitant displacement of SYN1/SCP1 and COR1/SCP3 from their chromosomes, do not have this capability. These data support the hypothesis that phosphorylation of SYN1/SCP1 and COR1/SCP3 targets their removal from the chromosomes and that activity of the kinases involved correlates with the presence of these two proteins on the chromosomes.


In situ infrared study of the role of PEG in stabilizing silica-supported amines for CO(2) capture.
The CO(2) capture capacity, adsorption mechanism, and degradation characteristics of two sorbents, silica-supported tetraethylenepentamine (TEPA/SiO(2)) and polyethylene-glycol-modified TEPA/SiO(2) (PEG/TEPA/SiO(2)), are studied by diffuse reflectance infrared Fourier transform spectroscopy and mass spectrometry. The CO(2) capture capacities of TEPA/SiO(2) and PEG/TEPA/SiO(2) are determined to be 2087 and 1110 micromol CO(2) g(-1) sorbent, respectively. Both sorbents adsorb CO(2) as hydrogen-bonding species, NH(2)--O, and carbamate/carboxylate species. The CO(2) adsorption half-time increases with the number of CO(2) capture cycles. Infrared results suggest that the increased adsorption half-time is a result of diffusion limitation, caused by accumulation of TEPA and PEG species on the surface of the sorbent particles. The degradation of TEPA/SiO(2) is found to correlate with the accumulation of carboxylate/carbamic species. The addition of PEG decreases the degradation rate of the sorbent and slows down the formation of carboxylate species. These carboxylate species can block CO(2) capture on amine (NH(2)/NH) sites. The stabilizing role of PEG on TEPA/SiO(2) can be attributed to hydrogen-bonding between TEPA (NH(2)/NH)and PEG (OH).


Early loss of neurogenic inflammation in the human diabetic foot.
1. Neurogenic inflammation, mediated by nociceptor C fibres, is part of the acute neurovascular response to injury producing the axon reflex flare. Laser Doppler flowmetry was used to measure the flare response induced by the electrophoresis, at various current strengths, of a ring of acetylcholine solution into dorsal foot skin. 2. Nineteen control subjects and 52 long-duration insulin-dependent (Type 1) diabetic patients of similar age (20 without complications; 19 with laser-treated retinopathy; 13 with reduced vibration perception and retinopathy) were studied in order to investigate the possible attenuation of this defence mechanism in diabetes. 3. The maximal (1 mA) flare response [control median (interquartile range): 1.55 (1.16-2.06) arbitrary units] was reduced greatly in neuropathic patients [0.37 (0.24-0.66) arbitrary units; P less than or equal to 0.001 with respect to all other groups], especially those with a previous history of foot ulceration. The flare was also reduced in some patients with retinopathy alone [1.06 (0.56-1.27) arbitrary units; P less than 0.005 with respect to control subjects]. 4. No rightward shift of the curve of hyperaemic response plotted against current strength was found, suggesting that the abnormal response was due to axonal loss rather than to dysfunction. 5. Neurogenic inflammation, mediated by small pain fibres, was markedly impaired in a group of diabetic patients at risk of foot ulceration. Furthermore, impairment of this nociceptor C fibre response can develop before clinical large-fibre neuropathy and could itself predispose to foot complications.


Membrane bridging by Munc13-1 is crucial for neurotransmitter release.
Munc13-1 plays a crucial role in neurotransmitter release. We recently proposed that the C-terminal region encompassing the C1, C2B, MUN and C2C domains of Munc13-1 (C1C2BMUNC2C) bridges the synaptic vesicle and plasma membranes through interactions involving the C2C domain and the C1-C2B region. However, the physiological relevance of this model has not been demonstrated. Here we show that C1C2BMUNC2C bridges membranes through opposite ends of its elongated structure. Mutations in putative membrane-binding sites of the C2C domain disrupt the ability of C1C2BMUNC2C to bridge liposomes and to mediate liposome fusion in vitro. These mutations lead to corresponding disruptive effects on synaptic vesicle docking, priming, and Ca2+-triggered neurotransmitter release in mouse neurons. Remarkably, these effects include an almost complete abrogation of release by a single residue substitution in this 200 kDa protein. These results show that bridging the synaptic vesicle and plasma membranes is a central function of Munc13-1.


Maternal uniparental isodisomy of chromosome 6 reveals a TULP1 mutation as a novel cause of cone dysfunction.
The majority of the genetic causes of autosomal recessive (ar) cone dystrophy (CD) and cone-rod dystrophy (CRD) are currently unknown. We used a high-resolution homozygosity mapping approach in a cohort of patients with CD or CRD to identify new genes for ar cone disorders. Case series. A cohort of 159 patients with ar CD and 91 patients with CRD. The genomes of 83 patients with ar CD and 73 patients with CRD were analyzed for homozygous regions using single nucleotide polymorphism (SNP) microarrays. One patient showed homozygosity of SNPs across chromosome 6, and segregation analysis was performed using microsatellite markers. Direct sequencing of all retinal disease genes on chromosome 6 revealed a novel pathogenic TULP1 mutation in this patient. A cohort of 159 individuals with CD and 91 individuals with CRD was screened for this particular mutation using the restriction enzyme HhaI. The medical history of patients carrying the TULP1 mutation was reviewed and additional ophthalmic examinations were performed, including electroretinography (ERG), perimetry, optical coherence tomography (OCT), fundus autofluorescence (FAF), and fundus photography. TULP1 mutations, age at diagnosis, visual acuity, fundus appearance, color vision defects, visual field, ERG, FAF, and OCT findings. In 1 patient, homozygosity mapping and subsequent segregation analysis revealed maternal uniparental disomy (UPD) of chromosome 6. A novel homozygous missense mutation (p.Arg420Ser) was identified in TULP1, whereas no mutations were detected in other retinal disease genes on chromosome 6. The mutation affects a highly conserved amino acid residue in the Tubby domain and is predicted to be pathogenic. The same homozygous mutation was also identified in an additional, unrelated patient with CRD. Both patients carrying the p.Arg420Ser mutation presented with a bull's eye maculopathy. The first patient had progressive loss of visual acuity with a relatively preserved ERG, whereas the second patient developed loss of visual acuity, peripheral degeneration, and severely reduced ERG responses in a cone-rod pattern. Maternal UPD of chromosome 6 unmasked a mutation in the TULP1 gene as a novel cause of cone dysfunction. This expands the disease spectrum of TULP1 mutations from Leber congenital amaurosis and early-onset retinitis pigmentosa to cone-dominated disease. The author(s) have no proprietary or commercial interest in any materials discussed in this article.


Tracking the luminal exposure and lymphatic drainage pathways of intravaginal and intrarectal inocula used in nonhuman primate models of HIV transmission.
Over 80% of sexual HIV-1 transmissions originate from a single viral variant, but the underlying basis for this transmission bottleneck remains to be elucidated. Nonhuman primate models of mucosal virus transmission allow opportunities to gain insight into the basis of this mucosal bottleneck. We used simulated inocula consisting of either non-infectious vital dye or contrast dye with non-invasive magnetic resonance imaging (MRI) to visualize mucosal exposure and passive lymphatic drainage patterns following vaginal and rectal exposures in Indian origin rhesus macaques. Results revealed a limited overall distance of dye coverage from the anal verge following 1 ml (n = 8) intrarectally administered, which greatly increased with a 3 ml (n = 8) volume. Intravaginal dye exposure using 2 ml revealed complete coverage of the mucosa of the vagina and ectocervix, however dye was not detectable in the endocervix, uterus, fallopian tubes or ovaries in nuliparous sexually mature rhesus macaques (n = 9). In addition, following submucosal and intranodal injections of vital dye or MRI contrast dye in the rectum (n = 9), or distal and proximal vagina (n = 4), the lymphatic drainage pathways were identified as first the internal then common iliac chain followed by para-aortic lymph nodes. Drainage from the distal descending colon (n = 8) was via the para-colonic lymph nodes followed by the inferior mesenteric and para-aortic lymph nodes. Analysis after vaginal challenge with infectious SIVmac239 followed by euthanasia at day 3 revealed a pattern of viral dissemination consistent with the imaging results. These results provide insights into potential patterns of viral dissemination that can help guide efforts to better elucidate the earliest events of virus transmission and potential intervention strategies.


Angiotensin-converting enzyme gene polymorphism and carotid wall thickening in a community population.
The insertion/deletion (I/D) polymorphism of the angiotensin-converting enzyme (ACE) gene has been associated with an increased risk of coronary heart disease, but whether it is a risk factor for underlying atherosclerosis remains unclear. Therefore, we examined to see whether the ACE gene deletion polymorphism was associated with carotid wall thickening and atherosclerotic plaque formation in a large randomly selected community population. A total of 1111 subjects, aged 27 to 77 years, with an equal male:female ratio and equal numbers in each age decile, were randomly selected from the Perth community population. Mean common carotid intima-medial wall thickness (IMT) and focal plaque formation were assessed by high-resolution B-mode ultrasound. The ACE gene I/D polymorphism was detected by PCR. The distribution of the ACE genotypes conformed to the Hardy-Weinberg equilibrium (DD, 31.0%; ID, 48.4%; and II, 20.6%). The D allele was strongly correlated in a codominant fashion with plasma ACE activity (r(s)=0.53, P<0.0001), and accounted for 33% of the total variance in circulating ACE activity. No significant differences among the ACE genotypes were found with respect to age, sex, and conventional risk variables, including a history of hypertension and vascular disease. The average mean IMT and prevalence of increased IMT and focal plaque were not significantly different among genotypes in the overall population or in the subset (n=852) who were conventionally low risk by Framingham coronary heart disease risk score. Logistic regression analysis selected age, systolic blood pressure, pack-years of smoking, LDL cholesterol level, waist/hip ratio, and history of hypertension, but not the D allele, as multivariate predictors of increased IMT and carotid plaque formation. We conclude that, although the ACE I/D polymorphism is strongly related to ACE activity, it is not a risk predictor of carotid wall thickening or focal plaque formation when examined in a large randomly selected community population.


SU-E-T-320: A New Verification Phantom for GYN Brachytherapy Applicators Using GafChromic - Films.
Brachytherapy plays an important role in radiation therapy a wide range of tumor sits such as vaginal, cervical and endometrial cancers. The purpose of this project was to design, fabricate and verify a new phantom for dosimetric verification at small distances from GYN applicators used with GZP6 cobalt-60 HDR system. A new phantom has been designed and fabricated from 90 slabs of 18×16×0.2 cm3 Perspex to accommodate one tandem and two ovoids. The thin layer of the slabs was chosen to place GafChromic films in between the slabs for dosimetry with GZP6 cobalt-60 HDR system. For verification of this device, an assembly composed of a large ovoid size (3cm diameter) and tandem #1 with the least curvature was selected in this study. With this assembly, GafChromic films were exposed using a plan with 500 cGy dose delivery to point "A". The irradiated films were scanned. The responses of the films were converted to dose by calibrating samples of these films using a cobalt-60 teletherapy system in the range of 25 to 800 cGy dose. The measured isodose curves with the films were compared to calculated isodose lines by the treatment planning software. The Result of these investigations indicated differences of up to ± 23 % between the planning and measured dosimetry at different points in GYN implant with cobalt-60 HDR source of GZP6 system. Therefore, this phantom enabled us to confirm the accuracy of radiation delivery to the GYN patients with cobalt-60 HDR source of GZP6 system. The new phantom design could be utilized for the QA procedure of the GZP6 cobalt-60 HDR system as well as the Ir-192 HDR system to confirmation the accuracy of dose distribution in GYN implants, especially in non-traditional implants. The Radiotherapy Department of Shahid Beheshti University at Shohada hospital sponsored the purchase of the phantom materials and films used in the investigations.


Topology Automated Force-Field Interactions (TAFFI): A Framework for Developing Transferable Force Fields.
Force-field development has undergone a revolution in the past decade with the proliferation of quantum chemistry based parametrizations and the introduction of machine learning approximations of the atomistic potential energy surface. Nevertheless, transferable force fields with broad coverage of organic chemical space remain necessary for applications in materials and chemical discovery where throughput, consistency, and computational cost are paramount. Here, we introduce a force-field development framework called Topology Automated Force-Field Interactions (TAFFI) for developing transferable force fields of varying complexity against an extensible database of quantum chemistry calculations. TAFFI formalizes the concept of atom typing and makes it the basis for generating systematic training data that maintains a one-to-one correspondence with force-field terms. This feature makes TAFFI arbitrarily extensible to new chemistries while maintaining internal consistency and transferability. As a demonstration of TAFFI, we have developed a fixed-charge force-field, TAFFI-gen, from scratch that includes coverage for common organic functional groups that is comparable to established transferable force fields. The performance of TAFFI-gen was benchmarked against OPLS and GAFF for reproducing several experimental properties of 87 organic liquids. The consistent performance of these force fields, despite their distinct origins, validates the TAFFI framework while also providing evidence of the representability limitations of fixed-charge force fields.


Compartmentalized calcium signaling triggers subpopulation formation upon platelet activation through PAR1.
Blood platelets need to undergo activation to carry out their function of stopping bleeding. Different activation degrees lead to a stepped hierarchy of responses: ability to aggregate, granule release, and, in a fraction of platelets, phosphatidylserine (PS) exposure. This suggests the existence of decision-making mechanisms in the platelet intracellular signaling network. To identify and investigate them, we developed a computational model of PAR1-stimulated platelet signal transduction that included a minimal set of major players in the calcium signaling network. The model comprised three intracellular compartments: cytosol, dense tubular system (DTS) and mitochondria and extracellular space. Computer simulations showed that the stable resting state of platelets is maintained via a balance between calcium pumps and leaks through the DTS and plasma membranes. Stimulation of PAR1 induced oscillations in the cytosolic calcium concentrations, in good agreement with experimental observations. Further increase in the agonist level activated the mitochondrial uniporter leading to calcium uptake by mitochondria, which caused the collapse of mitochondrial membrane potential in a fraction of platelets leading to the PS exposure. The formation of this subpopulation was shown to be a stochastic process determined by the small number of activated PAR1 receptors and by heterogeneity in the number of ion pumps. These results demonstrate how a gradual increase of the activation degree can be converted into a stepped response hierarchy ultimately leading to formation of two distinct subpopulations from an initially homogeneous population.


The report of Task Group 100 of the AAPM: Application of risk analysis methods to radiation therapy quality management.
The increasing complexity of modern radiation therapy planning and delivery challenges traditional prescriptive quality management (QM) methods, such as many of those included in guidelines published by organizations such as the AAPM, ASTRO, ACR, ESTRO, and IAEA. These prescriptive guidelines have traditionally focused on monitoring all aspects of the functional performance of radiotherapy (RT) equipment by comparing parameters against tolerances set at strict but achievable values. Many errors that occur in radiation oncology are not due to failures in devices and software; rather they are failures in workflow and process. A systematic understanding of the likelihood and clinical impact of possible failures throughout a course of radiotherapy is needed to direct limit QM resources efficiently to produce maximum safety and quality of patient care. Task Group 100 of the AAPM has taken a broad view of these issues and has developed a framework for designing QM activities, based on estimates of the probability of identified failures and their clinical outcome through the RT planning and delivery process. The Task Group has chosen a specific radiotherapy process required for "intensity modulated radiation therapy (IMRT)" as a case study. The goal of this work is to apply modern risk-based analysis techniques to this complex RT process in order to demonstrate to the RT community that such techniques may help identify more effective and efficient ways to enhance the safety and quality of our treatment processes. The task group generated by consensus an example quality management program strategy for the IMRT process performed at the institution of one of the authors. This report describes the methodology and nomenclature developed, presents the process maps, FMEAs, fault trees, and QM programs developed, and makes suggestions on how this information could be used in the clinic. The development and implementation of risk-assessment techniques will make radiation therapy safer and more efficient.


Antibiotic and Modulation of Microbiota: A New Paradigm?
Recently new insights on gut microbiota have revolutionized many concepts of the modern medicine. The alteration of microbiota, which is called dysbiosis, has been associated with an expanding list of diseases and conditions. The development of next-generation sequencing techniques allowed comprehensive analysis of gut microbiota composition without the limitations of classic culture methods. Furthermore, introduction of functional techniques such as metabolomics and proteomics allowed for integrated analysis thus obtaining more robust insights on microbiota functions in health and disease. These tools allow to address the role of factors able to modify the gut microbiota, the so called "microbiota influencers." These data are useful to explain the physiopathology of several disease and thus to identify new potential therapeutic targets. Among microbiota influencers, many studies focused on the impact of antibiotic administration on the gut microbiota, because of their widespread use. Notably, beside the known beneficial effect of antibiotic in treating infectious diseases, these drugs have shown detrimental effects on gut microbiota which, in turn, might have long-term consequences on the host. Finally, therapeutic modulation of gut microbiota, by means of selected antibiotics with eubiotic effects, probiotics and with fecal microbiota transplantation seems of great interest as it might be able to prevent or even revert antibiotic-induced dysbiosis.


Soil resources and element stocks in drylands to face global issues.
Drylands (hyperarid, arid, semiarid, and dry subhumid ecosystems) cover almost half of Earth's land surface and are highly vulnerable to environmental pressures. Here we provide an inventory of soil properties including carbon (C), nitrogen (N), and phosphorus (P) stocks within the current boundaries of drylands, aimed at serving as a benchmark in the face of future challenges including increased population, food security, desertification, and climate change. Aridity limits plant production and results in poorly developed soils, with coarse texture, low C:N and C:P, scarce organic matter, and high vulnerability to erosion. Dryland soils store 646 Pg of organic C to 2 m, the equivalent of 32% of the global soil organic C pool. The magnitude of the historic loss of C from dryland soils due to human land use and cover change and their typically low C:N and C:P suggest high potential to build up soil organic matter, but coarse soil textures may limit protection and stabilization processes. Restoring, preserving, and increasing soil organic matter in drylands may help slow down rising levels of atmospheric carbon dioxide by sequestering C, and is strongly needed to enhance food security and reduce the risk of land degradation and desertification.


Simulation of eolian saltation.
Saltation is important in the transport of sand-sized granular material by wind and in the ejection of dust from the bed both on Earth and on Mars. The evolution of the saltating population and all its characteristic profiles is calculated from inception by pure aerodynamic entrainment through to steady state. Results of numerical simulations of single-grain impacts into granular beds are condensed into analytic expressions for the number and speeds of grains rebounding or rejected (splashed) from the bed. A model is combined with (i) this numerical representation, (ii) an expression for the aerodynamic entrainment rate, and (iii) the modification of the wind velocity profile by saltating grains. Calculated steady state mass fluxes are within the range of mass fluxes measured in wind tunnel experiments; mass flux is nonlinearly dependent on the shear velocity. Aerodynamically entrained grains in the system are primarily seeding agents; at steady state, aerodynamic entrainment is rare. The time for the entire system to reach steady state is roughly 1 second, or several long-trajectory hop times.


Very low pandemic influenza A (H1N1) 2009 mortality associated with early neuraminidase inhibitor treatment in Japan: analysis of 1000 hospitalized children.
There were many cases of pandemic influenza A (H1N1) 2009 (H1N1/09) in Japan during the 2009-2010 epidemic. They accounted for 16% of the total population (20.7 million/128 million), and 59% of the patients were children 15 years of age and under (12.2 million/20.7million). However, there were only 38 paediatric deaths. We analyzed the clinical manifestations and treatment of children hospitalized because of H1N1/09 infection in order to clarify the association between treatment with neuraminidase inhibitors and the low mortality rate. A retrospective chart review was performed on a total of 1000 paediatric inpatients. The causes of the hospitalizations were respiratory complications in 651 cases (65.1%), neurological complications in 255 cases (25.5%) and other complications in 94 cases. Neuraminidase inhibitors, primarily oseltamivir, had been used to treat 984 (98.4%) of the 1000 patients, and in 88.9% of the patients, treatment with neuraminidase inhibitors was initiated within 48 h after the onset of illness. Only 12 (1.2%) of the 1000 patients underwent mechanical ventilation, and one patient died of H1N1/09 infection. Although a high proportion of the patients in this study had severe respiratory complications, the case fatality rate was only 0.1%. The low mortality rate of children due to the H1N1/09 epidemic in Japan was probably attributable to the universal implementation of early treatment with neuraminidase inhibitors.


Quantitative assessment of the importance of phenotypic plasticity in adaptation to climate change in wild bird populations.
Predictions about the fate of species or populations under climate change scenarios typically neglect adaptive evolution and phenotypic plasticity, the two major mechanisms by which organisms can adapt to changing local conditions. As a consequence, we have little understanding of the scope for organisms to track changing environments by in situ adaptation. Here, we use a detailed individual-specific long-term population study of great tits (Parus major) breeding in Wytham Woods, Oxford, UK to parameterise a mechanistic model and thus directly estimate the rate of environmental change to which in situ adaptation is possible. Using the effect of changes in early spring temperature on temporal synchrony between birds and a critical food resource, we focus in particular on the contribution of phenotypic plasticity to population persistence. Despite using conservative estimates for evolutionary and reproductive potential, our results suggest little risk of population extinction under projected local temperature change; however, this conclusion relies heavily on the extent to which phenotypic plasticity tracks the changing environment. Extrapolating the model to a broad range of life histories in birds suggests that the importance of phenotypic plasticity for adjustment to projected rates of temperature change increases with slower life histories, owing to lower evolutionary potential. Understanding the determinants and constraints on phenotypic plasticity in natural populations is thus crucial for characterising the risks that rapidly changing environments pose for the persistence of such populations.


BCR-ABL uncouples canonical JAK2-STAT5 signaling in chronic myeloid leukemia.
Constitutive activation of STAT5 is critical for the maintenance of chronic myeloid leukemia (CML) characterized by the BCR-ABL oncoprotein. Tyrosine kinase inhibitors (TKIs) for the STAT5-activating kinase JAK2 have been discussed as a treatment option for CML patients. Using murine leukemia models combined with inducible ablation of JAK2, we show JAK2 dependence for initial lymphoid transformation, which is lost once leukemia is established. In contrast, initial myeloid transformation and leukemia maintenance were independent of JAK2. Nevertheless, several JAK2 TKIs induced apoptosis in BCR-ABL(+) cells irrespective of the presence of JAK2. This is caused by the previously unknown direct 'off-target' inhibition of BCR-ABL. Cellular and enzymatic analyses suggest that BCR-ABL phosphorylates STAT5 directly. Our findings suggest uncoupling of the canonical JAK2-STAT5 module upon BCR-ABL expression, thereby making JAK2 targeting dispensable. Thus, attempts to pharmacologically target STAT5 in BCR-ABL(+) diseases need to focus on STAT5 itself.


Slb/Wnt11 controls hypoblast cell migration and morphogenesis at the onset of zebrafish gastrulation.
During vertebrate gastrulation, highly coordinated cellular rearrangements lead to the formation of the three germ layers, ectoderm, mesoderm and endoderm. In zebrafish, silberblick (slb)/wnt11 regulates normal gastrulation movements by activating a signalling pathway similar to the Frizzled-signalling pathway, which establishes epithelial planar cell polarity (PCP) in Drosophila. However, the cellular mechanisms by which slb/wnt11 functions during zebrafish gastrulation are still unclear. Using high-resolution two-photon confocal imaging followed by computer-assisted reconstruction and motion analysis, we have analysed the movement and morphology of individual cells in three dimensions during the course of gastrulation. We show that in slb-mutant embryos, hypoblast cells within the forming germ ring have slower, less directed migratory movements at the onset of gastrulation. These aberrant cell movements are accompanied by defects in the orientation of cellular processes along the individual movement directions of these cells. We conclude that slb/wnt11-mediated orientation of cellular processes plays a role in facilitating and stabilising movements of hypoblast cells in the germ ring, thereby pointing at a novel function of the slb/wnt11 signalling pathway for the regulation of migratory cell movements at early stages of gastrulation.


New mass spectrometry technologies contributing towards comprehensive and high throughput omics analyses of single cells.
Mass-spectrometry based omics technologies - namely proteomics, metabolomics and lipidomics - have enabled the molecular level systems biology investigation of organisms in unprecedented detail. There has been increasing interest for gaining a thorough, functional understanding of the biological consequences associated with cellular heterogeneity in a wide variety of research areas such as developmental biology, precision medicine, cancer research and microbiome science. Recent advances in mass spectrometry (MS) instrumentation and sample handling strategies are quickly making comprehensive omics analyses of single cells feasible, but key breakthroughs are still required to push through remaining bottlenecks. In this review, we discuss the challenges faced by single cell MS-based omics analyses and highlight recent technological advances that collectively can contribute to comprehensive and high throughput omics analyses in single cells. We provide a vision of the potential of integrating pioneering technologies such as Structures for Lossless Ion Manipulations (SLIM) for improved sensitivity and resolution, novel peptide identification tactics and standards free metabolomics approaches for future applications in single cell analysis.


The partial 5-hydroxytryptamine1A receptor agonist buspirone does not antagonize morphine-induced respiratory depression in humans.
Based on experiments in rats, serotonin receptor 5-hydroxytryptamine (5-HT)(1A) agonists have been proposed as a potential therapeutic strategy for the selective treatment of opioid-induced respiratory depression. We investigated the clinical applicability of this principle in healthy volunteers. Twelve subjects received 0.43 mg/kg morphine (30 mg for 70 kg body weight) administered intravenously (i.v.) over approximately 2 h. At the start of the morphine infusion, they received in a randomized, double-blind cross-over design 60 mg p.o. buspirone or placebo. Respiratory depression (hypercapnic challenge) and pain (electrical stimuli: 5 Hz sinus 0-20 mA; chemical stimuli: 200 ms gaseous CO(2) pulses applied to the nasal mucosa) were assessed at baseline, at the end of the morphine infusion, and a third time after antagonizing the opioid effects by i.v. administration of 2 mg naloxone. The linear relationship between the minute ventilation and the CO(2) concentration in the inspired air of 1.07+/-0.27 l/mm Hg CO(2) at baseline conditions became shallower (0.45+/-0.23 l/mm Hg CO(2)) after morphine administration (P<0.001), indicating respiratory depression, which was significantly reversed by naloxone (0.95+/-0.43 l/mm Hg CO(2); P=0.001). Co-administration of buspirone had no effect on morphine-induced respiratory depression (slope 0.45+/-0.23 l/mm Hg CO(2) under morphine plus placebo versus 0.38+/-0.25 l/mm Hg CO(2) under morphine plus buspirone; P=0.7). Significant morphine-induced analgesia was observed in both pain models and was reversed by naloxone but unaffected by buspirone. Buspirone significantly increased the nausea induced by morphine (P=0.011). Oral co-administration of a high dose of the clinically available 5-HT(1A) agonist buspirone cannot be advised as a remedy for opioid-induced respiratory depression. This is indicated by its lack of anti-respiratory depressive effects and by the buspirone-associated increase of morphine-induced nausea.


Osteonecrosis: An emerging complication of intensive chemotherapy for childhood acute lymphoblastic leukemia.
Osteonecrosis (ON) is a potentially disabling complication of combination chemotherapy including high doses of steroids. The incidence and main risk factors for symptomatic ON have been investigated in a large group of children treated with high-dose steroids, prednisone and dexamethasone for childhood acute lymphoblastic leukemia (ALL). From May 1995 to December 1999, 1421 patients <18 years old, with newly diagnosed non-B ALL, were registered in the AIEOP-ALL 95 study. Their data were reviewed to identify patients who developed symptomatic ON. For those who were positively identified additional data were requested concerning ON-related symptoms, treatment and outcome. Overall, 15 of the 1421 patients developed symptomatic ON (1.1%) in a total of 29 sites. The estimated 5-year cumulative risk for clinically diagnosed ON was 1.6% (SE 0.4). The incidence was significantly higher among females (p=0.01) and older patients, with a peak rate of 7.4% (2.3) among those aged 10 to 17 years (p<0.0001). When the two factors, i.e. age and gender were combined, there was a striking increase in the risk among female patients aged 10 to 17 years. The median time between the diagnosis of ALL and that of ON was 17 months (range 8-45). The hip was the most frequently involved (19/29) site. Symptomatic ON occurred in only 1.1% of patients treated with BFM-type, intensive chemotherapy for childhood ALL. Female adolescents appear to be the subset of patients with the highest risk of ON, especially when categorized as having high risk leukemia and thus administered higher cumulative doses of dexamethasone.


Inhibiting Heat Shock Proteins Can Potentiate the Cytotoxic Effect of Cannabidiol in Human Glioma Cells.
Cannabinoids possess a number of characteristics that make them putative anticancer drugs, and their value as such is currently being explored in a number of clinical studies. To further understand the roles that cannabinoids may have, we performed gene expression profiling in glioma cell lines cultured with cannabidiol (CBD) and/or Δ9-tetrahydrocannabinol (THC), and pursued targets identified by this screening. Results showed that a large number of genes belonging to the heat shock protein (HSP) super-family were up-regulated following treatment, specifically with CBD. Increases were observed both at the gene and protein levels and arose as a consequence of increased generation of ROS by CBD, and correlated with an increase in a number of HSP client proteins. Furthermore, increases impeded the cytotoxic effect of CBD; an effect that was improved by co-culture with pharmacalogical inhibitors of HSPs. Similarly, culturing glioma cells with CBD and HSP inhibitors increased radiosensitivity when compared to CBD-alone. Taken together, these data indicate that the cytotoxic effects of CBD can be diminished by HSPs that indirectly rise as a result of CBD use, and that the inclusion of HSP inhibitors in CBD treatment regimens can enhance the overall effect.


Physician consensus on preventability and predictability of readmissions based on standard case scenarios.
Policy makers struggle with unplanned readmissions as a quality indicator since integrating preventability in such indicators is difficult. Most studies on the preventability of readmissions questioned physicians whether they consider a given readmission to be preventable, from which conclusions on factors predicting preventable readmissions were derived. There is no literature on the interobserver agreement of physician judgement. To assess the degree of agreement among physicians regarding predictability and preventability of medical readmissions. An online survey based on eight real-life case scenarios was distributed to European physicians. Physicians were requested to rate from the first four (index admission) scenarios whether they expected these patients to be readmitted within 30 days (the predictability). The remaining four cases, describing a readmission, were used to assess the preventability. The main outcome was the degree of agreement among physicians determined using the intra class correlation coefficient (ICC). 526 European medical physicians completed the survey. Most physicians had internal medicine as primary specialism. The median years of clinical experience was 11. ICC for predictability of readmission was 0.67 (moderate to good) and ICC for preventability of readmission was 0.13 (poor). There was moderate to good agreement among physicians on the predictability of readmissions while agreement on preventability was poor. This study indicates that assessing preventability of readmissions based solely on the judgement of physicians is far from perfect. Current literature on the preventability of readmissions and conclusions derived on the basis of physician opinion should be interpreted with caution.


Recognition sites for norepinephrine uptake: regulation by neurotransmitter.
Recognition sites for the uptake of norepinephrine on adrenergic neurons in the brain and periphery were labeled with [3H]desipramine. The number of these uptake sites varied with the concentration of transmitter; depletion of norepinephrine with reserpine reduced the number of uptake sites, whereas increasing the concentration of norepinephrine induced by treatment with monoamine oxidase inhibitors raised the number of binding sites. These dynamic alterations in norepinephrine uptake recognition sites may regulate synaptic function homeostatically, providing less inactivation of reuptake when the synaptic concentration of the transmitter is low and increased inactivation when it is high.


The Alternaria alternata Melanin Biosynthesis Gene Restores Appressorial Melanization and Penetration of Cellulose Membranes in the Melanin-Deficient Albino Mutant of Colletotrichum lagenarium
Colletotrichum lagenarium and Alternaria alternata produce a dark pigment, melanin. The C. lagenarium PKS1 and A. alternata ALM genes are involved in polyketide synthesis in the melanin biosynthesis pathway. PKS1 encodes a type I polyketide synthase. For functional comparison of the ALM gene with the PKS1 gene, we examined whether the A. alternata ALM gene could restore melanin synthesis in C. lagenarium albino mutant (Pks1(-)). The ALM gene transformed the albino mutant (Pks1(-)) to melanin-producing phenotypes, designated CAL transformants. The pigment intensity of both melanized colonies and appressoria of CAL transformants was weaker than that of the wild type. Ultrastructural studies of the cell walls of appressoria demonstrated that CAL transformants formed an outer melanized layer, as did the wild type. On the other hand, the thin inner and middle layers were less electron-dense than those of the wild type. CAL transformants were able to penetrate cellulose membranes as effectively as the wild type. By contrast, the penetration frequency of CAL transformants on cucumber cotyledons was remarkably reduced compared to that of the wild type. During conidial germination, the PKS1 transcript accumulated de novo in both the wild-type and CAL transformants after the start of conidial incubation. On the other hand, ALM transcript accumulated in conidia of CAL transformants before the start of conidial incubation.


Taxonomic note: a proposal for reviewing the interpretation of the CAMP reaction between Listeria monocytogenes and Rhodococcus equi.
The discrepancies between the current description of the CAMP test between Listeria monocytogenes and Rhodococcus equi in the latest edition of Bergey's Manual of Determinative Bacteriology (L. monocytogenes is described as CAMP test negative with R. equi) and routine findings (positive reactions are usually described in many laboratories) make it advisable to review the current interpretation of the CAMP test to avoid confusion among people working in microbiological laboratories. Overall, 98.4% of the L. monocytogenes strains examined in this study, regardless of their source or the intensity of their hemolytic activity, displayed a synergic hemolytic reaction (CAMP phenomenon) with R. equi, indicating that L. monocytogenes can generally be considered CAMP positive with R. equi. We propose that L. monocytogenes, together with Listeria ivanovii, should be considered CAMP test positive with R. equi (circular or racket and semicircular or shovel shapes, respectively).


Medical Simulation as a Vital Adjunct to Identifying Clinical Life-Threatening Gaps in Austere Environments.
Maternal mortality and morbidity are major causes of death in low-resource countries, especially those in Sub-Saharan Africa. Healthcare workforce scarcities present in these locations result in poor perioperative care access and quality. These scarcities also limit the capacity for progressive development and enhancement of workforce training, and skills through continuing medical education. Newly available low-cost, in-situ simulation systems make it possible for a small cadre of trainers to use simulation to identify areas needing improvement and to rehearse best practice approaches, relevant to the context of target environments. Nurse anesthetists were recruited throughout Sierra Leone to participate in simulation-based obstetric anesthesia scenarios at the country's national referral maternity hospital. All subjects participated in a detailed computer assisted training program to familiarize themselves with the Universal Anesthesia Machine (UAM). An expert panel rated the morbidity/mortality risk of pre-identified critical incidents within the scenario via the Delphi process. Participant responses to critical incidents were observed during these scenarios. Participants had an obstetric anesthesia pretest and post-test as well as debrief sessions focused on reviewing the significance of critical incident responses observed during the scenario. 21 nurse anesthetists, (20% of anesthesia providers nationally) participated. Median age was 41 years and median experience practicing anesthesia was 3.5 years. Most participants (57.1%) were female, two-thirds (66.7%) performed obstetrics anesthesia daily but 57.1% had no experience using the UAM. During the simulation, participants were observed and assessed on critical incident responses for case preparation with a median score of 7 out of 13 points, anesthesia management with a median score of 10 out of 20 points and rapid sequence intubation with a median score of 3 out of 10 points. This study identified substantial risks to patient care and provides evidence to support the feasibility and value of in-situ simulation-based performance assessment for identifying critical gaps in safe anesthesia care in the low-resource settings. Further investigations may validate the impact and sustainability of simulation based training on skills transfer and retention among anesthesia providers low resource environments.


De novo and biallelic DEAF1 variants cause a phenotypic spectrum.
To investigate the effect of different DEAF1 variants on the phenotype of patients with autosomal dominant and recessive inheritance patterns and on DEAF1 activity in vitro. We assembled a cohort of 23 patients with de novo and biallelic DEAF1 variants, described the genotype-phenotype correlation, and investigated the differential effect of de novo and recessive variants on transcription assays using DEAF1 and Eif4g3 promoter luciferase constructs. The proportion of the most prevalent phenotypic features, including intellectual disability, speech delay, motor delay, autism, sleep disturbances, and a high pain threshold, were not significantly different in patients with biallelic and pathogenic de novo DEAF1 variants. However, microcephaly was exclusively observed in patients with recessive variants (p < 0.0001). We propose that different variants in the DEAF1 gene result in a phenotypic spectrum centered around neurodevelopmental delay. While a pathogenic de novo dominant variant would also incapacitate the product of the wild-type allele and result in a dominant-negative effect, a combination of two recessive variants would result in a partial loss of function. Because the clinical picture can be nonspecific, detailed phenotype information, segregation, and functional analysis are fundamental to determine the pathogenicity of novel variants and to improve the care of these patients.


Evaluation of Soft Tissue Landmark Reliability between Manual and Computerized Plotting Methods.
The aim of the study is to evaluate the reliability of soft tissue landmark identification between manual and digital plot-tings in both X and Y axes. A total of 50 pretreatment lateral cephalograms were selected from patients who reported for orthodontic treatment. The digital images of each cephalogram were imported directly into Dolphin software for onscreen digi-talization, while for manual tracing, images were printed using a compatible X-ray printer. After the images were standardized, and 10 commonly used soft tissue landmarks were plotted on each cephalogram by six different professional observers, the values obtained were plotted in X and Y axes. Intraclass correlation coefficient was used to determine the intrarater reliability for repeated landmark plotting obtained by both the methods. The evaluation for reliability of soft tissue landmark plottings in both manual and digital methods after subjecting it to interclass correlation showed a good reliability, which was nearing complete homogeneity in both X and Y axes, except for Y axis of throat point in manual plotting, which showed moderate reliability as a cephalometric variable. Intraclass correlation of soft tissue nasion had a moderate reliability along X axis. Soft tissue pogonion shows moderate reliability in Y axis. Throat point exhibited moderate reliability in X axis. The interclass correlation in X and Y axes shows high reliability in both hard tissue and soft tissue except for throat point in Y axis, when plotted manually. The intraclass correlation is more consistent and highly reliable for soft tissue landmarks and the hard tissue landmark identification is also consistent. The results obtained for manual and digital methods were almost similar, but the digital landmark plotting has an added advantage in archiving, retrieval, transmission, and can be enhanced during plotting of lateral cephalograms. Hence, the digital method of landmark plotting could be preferred for both daily use and research because of the advantages.


Timing of Voice Therapy: A Primary Investigation of Voice Outcomes for Surgical Benign Vocal Fold Lesion Patients.
This study aims to examine the role of timing of speech-language pathology intervention on outcomes for surgical patients with benign vocal fold lesions as measured by a patient self-reported scale (Voice Handicap Index [VHI]) and objective acoustic measures (jitter %, Dysphonia Severity Index, noise-to-harmonics ratio). For the purpose of this study, interventions were categorized into three groups: preoperative (preoperative counseling session followed by postoperative therapy), pre- and postoperative therapy (multiple therapy sessions preoperatively and postoperatively), and postoperative therapy alone. Retrospective review of a prospective disease-specific outcomes database. Subjects identified in the database consisted of 12 preoperative counseling participants (six male, six female), 11 pre- and postoperative therapy participants (three male, eight female), and eight postoperative therapy only participants (three male, five female). Preoperative and postoperative VHI, Dysphonia Severity Index, jitter, and noise-to-harmonics ratio scores were compared between groups using paired t tests with a multiple comparison Bonferroni correction (α = 0.016). Despite lack of statistically significant changes in acoustic measures, groups receiving preoperative intervention whether with postoperative therapy or not, had statistically significant improvements in their VHI total scores (P = 0.001 for preoperative only and P = 0.0002 for pre- and postoperative therapy). Patients receiving speech pathology services before surgery for benign vocal fold lesions with or without postoperative therapy demonstrated greater gains in their subjective view of vocal function and quality as measured by the VHI.


[A cephalometric, electromyographic and kinesiographic appraisal of a patient with mandibular prognathism and anterior openbite malocclusion before and after surgical orthodontic therapy: a case report].
A 19-year-old female patient with skeletal 3 and anterior openbite malocclusions was treated by a surgical orthodontic approach. A glossectomy was also performed. Cephalometric, electromyographic and kinesiographic records were taken before and after the treatment. The treatment results were as follows: 1. Esthetic improvement in facial profile and static alignment and intercuspation of teeth was obtained. 2. The dimension of the tongue with respect to that of the oral cavity proper measured on the lateral cephalograms revealed a value similar to that determined for the control data after the active treatment. 3. The proportion of reversed strokes with respect to the total chewing strokes increased at the completion of the active treatment both for the right- and the left-sided chewings, but it decreased during the retentive period. 4. At the initial stage, an earlier onset of the masseter muscle activity relative to the temporal muscle group was determined, while the temporalis muscles showed an earlier onset of activity in the postoperative phase. 5. The durations of chewing strokes became shortened in a postoperative phase both on the working and the balancing sides. This observation was particularly significant for the opening and the closing phases. In addition, the proportion of the duration of significant temporalis muscle activity with respect to that of the concomitant chewing stroke increased. 6. Anterior temporalis muscle and masseter muscle activity during clenching performance revealed a value similar to the control data. In summary, the current case suggests that the patients who receive surgical orthodontic treatment require sufficient time to obtain functional readaptation during the retentive period.


Modelling suicide and unemployment: a longitudinal analysis covering 63 countries, 2000-11.
As with previous economic downturns, there has been debate about an association between the 2008 economic crisis, rising unemployment, and suicide. Unemployment directly affects individuals' health and, unsurprisingly, studies have proposed an association between unemployment and suicide. However, a statistical model examining the relationship between unemployment and suicide by considering specific time trends among age-sex-country subgroups over wider world regions is still lacking. We aimed to enhance knowledge of the specific effect of unemployment on suicide by analysing global public data classified according to world regions. We retrospectively analysed public data for suicide, population, and economy from the WHO mortality database and the International Monetary Fund's world economic outlook database from 2000 to 2011. We selected 63 countries based on sample size and completeness of the respective data and extracted the information about four age groups and sex. To check stability of findings, we conducted an overall random coefficient model including all study countries and four additional models, each covering a different world region. Despite differences in the four world regions, the overall model, adjusted for the unemployment rate, showed that the annual relative risk of suicide decreased by 1·1% (95% CI 0·8-1·4) per year between 2000 and 2011. The best and most stable final model indicated that a higher suicide rate preceded a rise in unemployment (lagged by 6 months) and that the effect was non-linear with higher effects for lower baseline unemployment rates. In all world regions, the relative risk of suicide associated with unemployment was elevated by about 20-30% during the study period. Overall, 41,148 (95% CI 39,552-42,744) suicides were associated with unemployment in 2007 and 46,131 (44,292-47,970) in 2009, indicating 4983 excess suicides since the economic crisis in 2008. Suicides associated with unemployment totalled a nine-fold higher number of deaths than excess suicides attributed to the most recent economic crisis. Prevention strategies focused on the unemployed and on employment and its conditions are necessary not only in difficult times but also in times of stable economy. University of Zurich.


Cystic Fibrosis Foundation otolaryngology care multidisciplinary consensus recommendations.
Cystic fibrosis (CF) is a multisystem disease that often requires otolaryngology care. Individuals with CF commonly have chronic rhinosinusitis but also present with hearing loss and dysphonia. Given these manifestations of CF, otolaryngologists are frequently involved in the care of patients with CF; however, there is limited consensus on optimal management of sinonasal, otologic, and laryngologic symptoms. The Cystic Fibrosis Foundation convened a multidisciplinary team of otolaryngologists, pulmonologists, audiologists, pharmacists, a social worker, a nurse coordinator, a respiratory therapist, two adults with CF, and a caregiver of a child with CF to develop consensus recommendations. Workgroups developed draft recommendation statements based on a systematic literature review, and a ≥80% consensus was required for acceptance of each recommendation statement. The committee voted on 25 statements. Eleven statements were adopted recommending a treatment or intervention, while five statements were formulated recommending against a specific treatment or intervention. The committee recommended eight statements as an option for select patients in certain circumstances, and one statement did not reach consensus. These multidisciplinary consensus recommendations will help providers navigate decisions related to otolaryngology consultation, medical and surgical management of CF-CRS, hearing, and voice in individuals with CF. A collaborative and multidisciplinary approach is advocated to best care for our patients with CF. Future clinical research is needed utilizing standardized, validated outcomes with comprehensive reporting of patient outcome, effects of modulator therapies, and genetic characteristics to help continue to advance care, decrease morbidity, and improve the quality of life for individuals with CF.


Urinary prostasin: a possible biomarker for renal pressure natriuresis in black adolescents.
Prostasin is a membrane-bound/secretive serine protease interacting with aldosterone and the epithelial sodium channel in the kidney. We and others have previously proposed the concept of stress-induced pressure natriuresis (SIPN) where increased urinary sodium excretion (UNaV) is coupled with elevated blood pressure (BP) in response to behavioral stress in normotensive adolescents. This study thus aimed to test the relationship between prostasin and pressure natriuresis using the SIPN model. A cohort of 102 normotensive black adolescents (mean age: 17.0 +/- 1.2 y; 56% females) were placed on a controlled sodium (4000 +/- 200 mg/d) and potassium (2600 +/- 200 mg/d) diet for three days before testing. The SIPN protocol consisted of a 1-h baseline period, a 1-h stress period (competitive video game), and a 1-h recovery period. During the stress period, BP elevation was coupled with an increase in UNaV. Urinary prostasin concentration had more than a 2-fold reduction from baseline (38.4 +/- 32.7 ng/mL) to stress (17.2 +/- 16.0 ng/mL), and further declined during recovery (12.1 +/- 16.2 ng/mL) (p < 0.001). Urinary prostasin was inversely correlated with UNaV during stress (r = -0.43, p = 0.0001), even after being normalized by urinary creatinine. Our data suggest that urinary prostasin could be a novel biomarker and/or mechanism for renal pressure natriuresis in normotensive black adolescents.


Fecal metabolomics of healthy breast-fed versus formula-fed infants before and during in vitro batch culture fermentation.
Nontargeted metabolomics analyses were used (1) to compare fecal metabolite profiles of healthy breast-fed (BF) and formula-fed (FF) infants before and during in vitro fermentation in batch culture and (2) to evaluate fecal metabolomics in assessing infant diet. Samples from healthy BF (n = 4) or FF (n = 4) infants were individually incubated at 37( °)C in anaerobic media containing 1% (wt/vol) galactooligosaccharides, 6'-sialyllactose, 2'-fucosyllactose, lacto-N-neotetraose, inulin, and gum arabic for up to 6 h, and supernatants were analyzed using GC/MS and LC/MS/MS to assess changes in various compounds. Comparison of over 250 metabolites prior to incubation showed that BF samples contained higher relative concentrations (P ≤ 0.05) of 14 compounds including human milk oligosaccharides and other metabolites presumably transferred through breast feeding (linoelaidate, myo-inositol) (P ≤ 0.05). Conversely, feces from FF infants contained 41 identified metabolites at higher levels (P ≤ 0.05) with many indicative of carbon limitation and protein fermentation. Our data are consistent with the notion that carbon-limited cultures catabolize protein and amino acids to obtain energy, whereas the provision of fermentable carbohydrate creates anabolic conditions relying on amino acids for bacterial growth. Results also suggest that fecal metabolomics can be a useful tool for studying interactions among diet, microbes, and host.


The impact of HIV antiviral therapy on human papillomavirus (HPV) infections and HPV-related diseases.
Human papillomavirus (HPV) infections play an important role in the pathogenesis of anogenital cancer and its precursors. HIV-infected individuals exhibit a high prevalence of HPV DNA. Several studies have further shown that HIV-infected individuals have an increased prevalence of squamous intraepithelial lesions (SIL) of the cervix, vulva and anus. The incidence of invasive cervical cancer is also elevated in HIV-positive women as well as that of anal cancer in HIV-positive women and men. Given the relationship between HIV-induced immunosuppression and HPV-associated disease, treatment with highly active antiretroviral therapy (HAART) has the potential, through immune reconstitution of the host, to alter the natural history of HPV infection and SIL. However, data on the impact of HAART on HPV disease are sparse and mixed results have been reported.


Enhanced susceptibility to lipopolysaccharide-induced arthritis and endotoxin shock in interleukin-32 alpha transgenic mice through induction of tumor necrosis factor alpha.
The present study assessed the potential functions of interleukin (IL)-32α on inflammatory arthritis and endotoxin shock models using IL-32α transgenic (Tg) mice. The potential signaling pathway for the IL-32-tumor necrosis factor (TNF)α axis was analyzed in vitro. IL-32α Tg mice were generated under control of a ubiquitous promoter. Two disease models were used to examine in vivo effects of overexpressed IL-32α: Toll-like receptor (TLR) ligand-induced arthritis developed using a single injection of lipopolysaccharide (LPS) or zymosan into the knee joints; and endotoxin shock induced with intraperitoneal injection of LPS and D-galactosamine. TNFα antagonist etanercept was administered simultaneously with LPS in some mice. Using RAW264.7 cells, in vitro effects of exogenous IL-32α on TNFα, IL-6 or macrophage inflammatory protein 2 (MIP-2) production were assessed with or without inhibitors for nuclear factor kappa B (NFκB) or mitogen-activated protein kinase (MAPK). Single injection of LPS, but not zymosan, resulted in development of severe synovitis with substantial articular cartilage degradation in knees of the Tg mice. The expression of TNFα mRNA in inflamed synovia was highly upregulated in the LPS-injected Tg mice. Moreover, the Tg mice were more susceptive to endotoxin-induced lethality than the wild-type control mice 48 hours after LPS challenge; but blockade of TNFα by etanercept protected from endotoxin lethality. In cultured bone marrow cells derived from the Tg mice, overexpressed IL-32α accelerated production of TNFα upon stimulation with LPS. Of note, exogenously added IL-32α alone stimulated RAW264.7 cells to express TNFα, IL-6, and MIP-2 mRNAs. Particularly, IL-32α -induced TNFα, but not IL-6 or MIP-2, was inhibited by dehydroxymethylepoxyquinomicin (DHMEQ) and U0126, which are specific inhibitors of nuclear factor kappa B (NFκB) and extracellular signal regulated kinase1/2 (ERK1/2), respectively. These results show that IL-32α contributed to the development of inflammatory arthritis and endotoxin lethality. Stimulation of TLR signaling with LPS appeared indispensable for activating the IL-32α-TNFα axis in vivo. However, IL-32α alone induced TNFα production in RAW264.7 cells through phosphorylation of inhibitor kappa B (IκB) and ERK1/2 MAPK. Further studies on the potential involvement of IL-32α-TNFα axis will be beneficial in better understanding the pathology of autoimmune-related arthritis and infectious immunity.


A synthon approach to artificial chemistry.
A coevolutionary model is discussed that incorporates the logical structure of constitutional chemistry and its kinetics on the one hand and the topological evolution of the chemical reaction network on the other hand. The motivation for designing this model is twofold. First, experiments that are to provide insight into chemical problems should be expressed in a syntax that remains as close as possible to real chemistry. Second, the study of physical properties of the complex chemical reaction networks requires growing models that incorporate features realistic from a biochemical perspective. In this article the theory and algorithms underlying the coevolutionary model are explained, and two illustrative examples are provided. These examples show that one needs to be careful in making general claims concerning the structure of chemical reaction networks.


A practical approach to the prevention of miscarriage: Part 3--Passive immunotherapy.
To evaluate the efficacy of passive immunotherapy in preventing miscarriage. Studies both pro and con concerning intravenous immunoglobulin therapy (IVIG) in preventing miscarriage were evaluated. A new therapy of i.v. intralipid infusion is also reviewed. Intravenous immunoglobulin therapy may be effective but it is necessary to use it prior to conception and monthly thereafter. Some brands are more potent than others. The data concerning intralipid i.v. infusion involves only small case series but the results from one study were encouraging though we could not personally substantiate these findings. Intravenous immunoglobulin therapy is very expensive. In the author's opinion there are no immunological studies that can determine if a woman needs immune suppression. The best way to decide is the history--the more miscarriages without any other identifiable cause the more likely passive immunotherapy may be helpful. If intralipid proves as efficacious as IVIG it will be a lot less expensive.


Heat-shock proteins during growth and sporulation of Bacillus subtilis.
Four major heat-shock proteins (hsps) with apparent molecular masses of 84, 69, 32 and 22 kDa were detected in exponentially growing stationary phase and sporulating cells of Bacillus subtilis heat-shocked from 30 to 43 degrees C. The most abundant, hsp69, is probably analogous to the E. coli groEL protein. These proteins were transiently inducible by heat-shock. Partial purification of RNA polymerase revealed several other minor hsps. One of these, a 48 kDa polypeptide probably corresponds to sigma 43. The synthesis of this polypeptide and at least two other proteins appeared to be under sporulation and heat-shock regulation and was affected by the SpoOA mutation.


Radioisotopes for metastatic bone pain.
This is an update of the review published in Issue 4, 2003. Bone metastasis cause severe pain as well as pathological fractures, hypercalcaemia and spinal cord compression. Treatment strategies currently available to relieve pain from bone metastases include analgesia, radiotherapy, surgery, chemotherapy, hormone therapy, radioisotopes and bisphosphonates. To determine efficacy and safety of radioisotopes in patients with bone metastases to improve metastatic pain, decrease number of complications due to bone metastases and improve patient survival. We sought randomised controlled trials (RCTs) in MEDLINE, EMBASE, CENTRAL, and the PaPaS Trials Register up to October 2010. Studies selected had metastatic bone pain as a major outcome after treatment with a radioisotope, compared with placebo or another radioisotope. We assessed the risk of bias of included studies by their sequence generation, allocation concealment, blinding of study participants, researchers and outcome assessors, and incomplete outcome data. Two review authors extracted data. We performed statistical analysis as an "available case" analysis, and calculated global estimates of effect using a random-effects model. We also performed an intention-to-treat (ITT) sensitivity analysis. This update includes 15 studies (1146 analyzed participants): four (325 participants) already included and 11 new (821 participants). Only three studies had a low risk of bias. We observed a small benefit of radioisotopes for complete relief (risk ratio (RR) 2.10, 95% CI 1.32 to 3.35; Number needed to treat to benefit (NNT) = 5) and complete/partial relief (RR 1.72, 95% CI 1.13 to 2.63; NNT = 4) in the short and medium term (eight studies, 499 participants). There is no conclusive evidence to demonstrate that radioisotopes modify the use of analgesia with respect to placebo. Leucocytopenia and thrombocytopenia are secondary effects significantly associated with the administration of radioisotopes (RR 5.03; 95% CI 1.35 to 18.70; Number needed to treat to harm (NNH) = 13). Pain flares were not higher in the radioisotopes group (RR 0.74; 95% CI 0.27 to 2.06). There are scarce data of moderate quality when comparing Strontium-89 ((89)Sr) with Samarium-153 ((153)Sm), Rhenium-186 ((186)Re) and Phosphorus-32 ((32)P). We observed no significant differences between treatments. Similarly, we observed no differences when we compared different doses of (153)Sm (0.5 versus 1.0 mCi). This update adds new evidence on efficacy of radioisotopes versus placebo, (89)Sr compared with other radioisotopes, and dose-comparisons of (153)Sm and (188)Re. There is some evidence indicating that radioisotopes may provide complete reduction in pain over one to six months with no increase in analgesic use, but severe adverse effects (leucocytopenia and thrombocytopenia) are frequent.


Incidentally-discovered adrenal masses.
Unanticipated adrenal masses are frequently encountered in modern, high resolution diagnostic imaging. Most often, these masses are benign adrenal adenomas, but when detected they necessitate a clinical evaluation sufficient to exclude subclinical endocrine disease, primary adrenal cancer, and remote metastases to the adrenal glands from other malignancies. These "incidentally-discovered" adrenal masses or so-called "adrenal incidentalomas" can be further evaluated with CT, MRI, and nuclear medicine imaging techniques. A substantial literature supports the use of each of these modalities to non-invasively characterize these neoplasms that have been considered by some as a 'disease' of modern imaging technology.


Eosinophilic esophagitis: updated consensus recommendations for children and adults.
Eosinophilic esophagitis (EoE) is a clinicopathologic condition of increasing recognition and prevalence. In 2007, a consensus recommendation provided clinical and histopathologic guidance for the diagnosis and treatment of EoE; however, only a minority of physicians use the 2007 guidelines, which require fulfillment of both histologic and clinical features. Since 2007, the number of EoE publications has doubled, providing new disease insight. Accordingly, a panel of 33 physicians with expertise in pediatric and adult allergy/immunology, gastroenterology, and pathology conducted a systematic review of the EoE literature (since September 2006) using electronic databases. Based on the literature review and expertise of the panel, information and recommendations were provided in each of the following areas of EoE: diagnostics, genetics, allergy testing, therapeutics, and disease complications. Because accumulating animal and human data have provided evidence that EoE appears to be an antigen-driven immunologic process that involves multiple pathogenic pathways, a new conceptual definition is proposed highlighting that EoE represents a chronic, immune/antigen-mediated disease characterized clinically by symptoms related to esophageal dysfunction and histologically by eosinophil-predominant inflammation. The diagnostic guidelines continue to define EoE as an isolated chronic disorder of the esophagus diagnosed by the need of both clinical and pathologic features. Patients commonly have high rates of concurrent allergic diatheses, especially food sensitization, compared with the general population. Proved therapeutic options include chronic dietary elimination, topical corticosteroids, and esophageal dilation. Important additions since 2007 include genetic underpinnings that implicate EoE susceptibility caused by polymorphisms in the thymic stromal lymphopoietin protein gene and the description of a new potential disease phenotype, proton pump inhibitor-responsive esophageal eosinophila. Further advances and controversies regarding diagnostic methods, surrogate disease markers, allergy testing, and treatment approaches are discussed.


Analysis of MDM2 and MDM4 single nucleotide polymorphisms, mRNA splicing and protein expression in retinoblastoma.
Retinoblastoma is a childhood cancer of the developing retina that begins in utero and is diagnosed in the first years of life. Biallelic RB1 gene inactivation is the initiating genetic lesion in retinoblastoma. The p53 gene is intact in human retinoblastoma but the pathway is believed to be suppressed by increased expression of MDM4 (MDMX) and MDM2. Here we quantify the expression of MDM4 and MDM2 mRNA and protein in human fetal retinae, primary retinoblastomas, retinoblastoma cell lines and several independent orthotopic retinoblastoma xenografts. We found that MDM4 is the major p53 antagonist expressed in retinoblastoma and in the developing human retina. We also discovered that MDM4 protein steady state levels are much higher in retinoblastoma than in human fetal retinae. This increase would not have been predicted based on the mRNA levels. We explored several possible post-transcriptional mechanisms that may contribute to the elevated levels of MDM4 protein. A proportion of MDM4 transcripts are alternatively spliced to produce protein products that are reported to be more stable and oncogenic. We also discovered that a microRNA predicted to target MDM4 (miR191) was downregulated in retinoblastoma relative to human fetal retinae and a subset of samples had somatic mutations that eliminated the miR-191 binding site in the MDM4 mRNA. Taken together, these data suggest that post-transcriptional mechanisms may contribute to stabilization of the MDM4 protein in retinoblastoma.


17 beta-Estradiol prevents dysfunction of canine coronary endothelium and myocardium and reperfusion arrhythmias after brief ischemia/reperfusion.
Brief myocardial ischemia is associated with myocardial and coronary endothelial dysfunction caused by oxygen free radicals released during reperfusion. Estrogen, known to have antioxidant activity, may prevent these complications. We assessed the effect of 2 weeks of treatment with 17 beta-estradiol (E, 100 micrograms.kg-1.d-1, n = 12) or placebo (P, n = 15) on myocardial and coronary endothelial function during the first 2 hours of reperfusion in dogs subjected to 15 minutes of ischemia induced by occlusion of the left anterior descending coronary artery (LAD). Our results show that the incidence of ventricular arrhythmias significantly decreased in E (3 of 12) compared with P (11 of 15). Systolic shortening, significantly depressed in P during early reperfusion, was maintained at preischemic levels in E. During reperfusion, the increase in LAD flow to acetylcholine, attenuated in P (60 +/- 6%), was preserved in E endothelium. (151 +/- 28%) and was associated with increased serum nitrite/nitrate concentration. n-Pentane in exhaled gas in vivo, an index of lipid peroxidation, increased significantly during early reperfusion in P (from 9.1 +/- 1.9 to 41.6 +/- 13.0 ppb, P < .05) but not in E (23.0 +/- 6.9 ppb). In vitro, arterial segments from E generated significantly less superoxide anion after hypoxia/reoxygenation than those from P. Ischemic/reperfused LAD segments from E also revealed a better preservation of endothelium-dependent relaxation in vitro (maximum relaxation, 42 +/- 4% versus 24 +/- 4% in P; P < .05). Estrogen protects against endothelial and myocardial dysfunction resulting from brief ischemia/reperfusion. This protection may relate to an antioxidant effect of estrogen.


Acanthosis Nigricansin PCOS Patients and Its Relation with Type 2 Diabetes Mellitus and Body Mass at a Tertiary Care Hospital in Southern India.
Acanthosis nigricansis was viewed recently as a possible marker of an increased risk for diabetes. Acanthosis Nigricans (AN) can help in identifying the patients with an increased risk for Type 2 Diabetes Mellitus (T2DM) among Polycystic Ovary Syndrome (PCOS) cases. Hence, this study was carried to know the prevalence of AN in PCOS and its correlation with diabetes mellitus and the body mass. A prospective, longitudinal study in the patients with PCOS, who attended the Department of Medicine, Kasturba Medical College [KMC] Hospital, Attavar, Mangalore, Karnatka, India between December 2008 to April 2010, was carried out. A total of 119 patients with PCOS were included in the study. A complete history, along with the demographic data of the patients who were aged between 15-35 years, were considered for the analysis. AN was observed in 56% of the PCOS patients. The Body Mass Index (BMI) and the waist circumference were significantly higher in the PCOS patients with AN, as compared to those without AN. Among the 119 PCOS patients,77% were found to have BMI values which were above the normal. Among the obese PCOS patients, 58% had BMI values of more than 30kg/m(2) as compared to 19% of the patients who did not have AN. Type 2 DM was observed in 77% of the PCOS patients with AN, while none of the PCOS patients were found to be diabetic among those without AN. A family history of DM in the first degree relatives was present in all the patients with AN. The results were analyzed by the Mann Whitney U test for the continuous data and by the Chi square test for the categorical data. The PCOS patients who had a family history of diabetes and obesity with a body mass index of more than 30 kg/m(2) and a waist circumference of ≥ 90 cm were more likely to develop AN, which is a marker of hyperinsulinaemia and insulin resistance. Identifying such PCOS patients will stimulate the discussions of the lifestyle modifications in the primary care setting.


The informational properties of the throwing arm for anticipation of goal-directed action.
We examined the informational value of biological motion from the arm in predicting the location of a thrown ball. In three experiments, participants were classified as being skilled and less skilled based on their actual performance on the task (i.e., using a within-task criterion). We then presented participants with a range of stick figure representations and required them to predict throw direction. In Experiment 1, we presented stick figure movies of a full body throwing action, right throwing arm plus left shoulder and throwing arm only. Participants were able to anticipate throw direction above chance under all conditions irrespective of perceptual skill level, with the perceptually skilled participants excelling under full body conditions. In Experiment 2, we neutralized dynamical differences in motion to opposing throw directions from the shoulder, elbow and wrist of the throwing arm. Neutralizing the wrist location negatively affected anticipation performance in all participants reducing accuracy to below chance. In Experiment 3, we presented movies of the motion wrist location alone and the upper section of the throwing arm (shoulder-elbow). Participants were able to successfully anticipate above chance in these latter two conditions. Our findings suggest that motion of the throwing arm contains multiple sources of information that can help facilitate the anticipation of goal-directed action. Perceptually skilled participants were superior in extracting informational value from motion at both the local and global levels when compared to less skilled counterparts.