Hypophosphataemic osteomalacia and myopathy: studies with nuclear magnetic resonance spectroscopy.
A patient with familial adult-onset hypophosphataemia, whose myopathy was closely related to the plasma phosphate concentration, was investigated by phosphorus nuclear magnetic resonance spectroscopy (31P n.m.r.) in vivo of the right flexor digitorum superficialis muscle. During hypophosphataemia induced by stopping oral phosphate a significant reduction in measured muscle strength occurred, but the ratios of the intramyocellular levels of phosphocreatine (PCr), adenosine triphosphate (ATP) and inorganic phosphate (Pi) remained unchanged at rest. During exercise these levels changed, as did the intramyocellular pH, but they did not differ from the pattern previously recorded in normal subjects. In four adults with inherited infantile-onset hypophosphataemia (vitamin D-resistant rickets, VDRR) without myopathy, the n.m.r. measurements were normal at rest and during exercise. In one patient with inherited hyperphosphataemia (tumoral calcinosis) the resting PCr: Pi ratio was significantly reduced.


Relaxations and phase transitions during the collapse of a dense PNIPAM microgel suspension-thorough insight using dielectric spectroscopy.
The dielectric behavior of a thermo-sensitive poly-(N-isopropylacrylamide) (PNIPAM) microgel suspension with a dense concentration was investigated over the frequency range of 40 Hz to 110 MHz in a wide temperature window of 10-60 °C. By successfully removing the electrode polarization effect from the original data, two remarkable and temperature-dependent relaxation processes were observed. Both of the two-phase transition processes, i.e., the colloidal crystal-to-liquid transition, which has not yet been detected by dielectric spectroscopy before, as well as the volume phase transition, were detected by the relaxation parameters. Based on the three physical states of the microgel suspension, the relaxation mechanisms are discussed in detail. The slow relaxation originates from the segmental motion and the counterion motion along the polymer chain over the whole temperature range. It was found that when the system is in the colloidal crystal and liquid state, the segmental motion is cooperative with side chain and hydrogen bonding networks, while in the phase separation state (at temperatures above the lower critical solution temperature (LCST)), the cooperative interaction disappears. The fast relaxation is due to the fluctuation of counterions below the LCST and the interfacial polarization above the LCST. Based on interfacial polarization theory, which describes the dielectric model of a conventional particle dispersion, the temperature dependence of the electrical properties for the constituent phases (the permittivity, conductivity and volume fraction of the microgel (εp, κp, ϕ); the conductivity of the medium water (κa); the water content in the PNIPAM microgel (fw)) were calculated using the Hanai equation. The water content is close to the result obtained using light scattering, indicating that the dielectric model for a conventional particle dispersion is also applicable to a soft atypical colloidal dispersion.


Molecular analysis and comparison of radiation-induced large deletions of the HPRT locus in primary human skin fibroblasts.
Genetic alterations in gamma-ray- and alpha-particle-induced HPRT mutants were examined by multiplex polymerase chain reaction (PCR) analysis. A total of 39-63% of gamma-ray-induced and 31-57% of alpha-particle-induced mutants had partial or total deletions of the HPRT gene. The proportion of these deletion events was dependent on radiation dose, and at the resolution limits employed there were no significant differences between the spectra induced by equitoxic doses of alpha particles (0.2-0.4 Gy) and gamma rays (3 Gy). The molecular nature of the deletions was analyzed by the use of sequence tagged site (STS) primers and PCR amplification as a "probe" for specific regions of the human X chromosome within the Xq26 region. These STSs were closely linked and spanned regions approximately 1.7 Mbp from the telomeric side and 1.7 Mbp from the centromeric side of the HPRT gene. These markers include: DXS53, 299R, DXS79, yH3L, 3/19, PR1, PR25, H2, yH3R, 1/44, 1/67, 1/1, DXS86, D8C6, DXS10 and DXS144. STS analyses indicated that the maximum size of total deletions in radiation-induced HPRT mutants can be greater than 2.7 Mbp and deletion size appears to be dependent on radiation dose. There were no apparent differences in the sizes of the deletions induced by alpha particles or gamma rays. On the other hand, deletions containing portions of the HPRT gene were observed to be 800 kbp or less, and the pattern of the partial deletion induced by alpha particles appeared to be different from that induced by gamma rays.


Exacerbated metastatic disease in a mouse mammary tumor model following latent gammaherpesvirus infection.
Controversy exists as to the ability of human gammaherpesviruses to cause or exacerbate breast cancer disease in patients. The difficulty in conducting definitive human studies can be overcome by investigating developing breast cancer in a mouse model. In this study, we utilized mice latently infected with murine gammaherpesvirus 68 (HV-68) to question whether such a viral burden could exacerbate metastatic breast cancer disease using a mouse mammary tumor model. Mice latently infected with HV-68 had a similar primary tumor burden, but much greater metastatic disease, when compared to mock treated mice given the transplantable tumor, 4 T1. This was true for lung lesions, as well as secondary tumor masses. Increased expression of pan-cytokeratin and VEGF-A in tumors from HV-68 infected mice was consistent with increased metastatic disease in these animals. Surprisingly, no viral particles could be cultured from tumor tissues, and the presence of viral DNA or RNA transcripts could not be detected in primary or secondary tumor tissues. Latent HV-68 infection had no significant effect on the size of primary 4 T1 mammary tumors, but exacerbated the number of metastatic lung lesions and secondary tumors when compared to mock treated mice. Increased expression of the tumor marker, pan-cytokeratin, and VEGF-A in tumors of mice harboring latent virus was consistent with an exacerbated metastatic disease. Mechanisms responsible for this exacerbation are indirect, since no virus could be detected in cancerous tissues.


PTSD symptoms and alcohol-related problems among veterans: Temporal associations and vulnerability.
Posttraumatic stress disorder (PTSD) is associated with elevated risk of both alcohol use disorder (AUD) and related conduct problems, which are associated with behavioral and emotional dysregulation. We conducted an intensive longitudinal burst design study with 10 weeks of experience sampling over the course of 1.5 years with 250 veterans of recent conflicts. We tested time-series models of daily associations between posttraumatic stress symptoms (PTSS), alcohol dependence syndrome, and conduct problems. Exacerbations of PTSS predicted higher dependence syndrome and conduct problems the next day. This effect was significant after controlling for both concurrent (i.e., same-day) associations between drinking and the outcomes as well as the strength of associations between the outcomes from one day to the next (i.e., autoregression). Affect lability and disinhibition were hypothesized vulnerability factors increasing the strength of within-person predictors of dependence syndrome and conduct problems. Lability and disinhibition were associated with greater dependence syndrome symptoms and conduct problems over the follow-up period. Consistent with expectation, lability rather than disinhibition increased the association between drinking and dependence syndrome as well as the strength of association between dependence syndrome symptoms from one day to the next. Moderating effects of disinhibition in the conduct problems model were not significant. Importantly, results indicated reciprocal associations over time. Lability potentiated the association between dependence syndrome symptoms and next day PTSS, whereas disinhibition potentiated the association between conduct problems and next day PTSS. Results demonstrate complex dynamic associations between PTSS, AUD symptoms, and conduct problems over time indicative of broad regulatory impairments. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Dependence of thymus development on derivatives of the neural crest.
Elimination of limited areas of the cephalic neural crest in stage 9 or 10 chick embryos markedly reduced the size of the thymus gland or resulted in its absence. Small thymic lobes contained both thymocytes and epithelial cells but showed delayed development. Parathyroid and thyroid glands sometimes were reduced in size or missing from the normal location on one or both sides. Heart defects were consistently present. Thymus development may depend on direct interaction of mesenchymal derivatives of the neural crest with pharyngeal epithelium. Multiple defects, such as the Di George syndrome, may result from failure of neural crest derivatives to migrate and interact properly.


Molecular modeling and in silico characterization of Mycobacterium tuberculosis TlyA: possible misannotation of this tubercle bacilli-hemolysin.
The TlyA protein has a controversial function as a virulence factor in Mycobacterium tuberculosis (M. tuberculosis). At present, its dual activity as hemolysin and RNA methyltransferase in M. tuberculosis has been indirectly proposed based on in vitro results. There is no evidence however for TlyA relevance in the survival of tubercle bacilli inside host cells or whether both activities are functionally linked. A thorough analysis of structure prediction for this mycobacterial protein in this study shows the need for reevaluating TlyA's function in virulence. Bioinformatics analysis of TlyA identified a ribosomal protein binding domain (S4 domain), located between residues 5 and 68 as well as an FtsJ-like methyltranferase domain encompassing residues 62 and 247, all of which have been previously described in translation machinery-associated proteins. Subcellular localization prediction showed that TlyA lacks a signal peptide and its hydrophobicity profile showed no evidence of transmembrane helices. These findings suggested that it may not be attached to the membrane, which is consistent with a cytoplasmic localization. Three-dimensional modeling of TlyA showed a consensus structure, having a common core formed by a six-stranded β-sheet between two α-helix layers, which is consistent with an RNA methyltransferase structure. Phylogenetic analyses showed high conservation of the tlyA gene among Mycobacterium species. Additionally, the nucleotide substitution rates suggested purifying selection during tlyA gene evolution and the absence of a common ancestor between TlyA proteins and bacterial pore-forming proteins. Altogether, our manual in silico curation suggested that TlyA is involved in ribosomal biogenesis and that there is a functional annotation error regarding this protein family in several microbial and plant genomes, including the M. tuberculosis genome.


Galantamine ameliorates the impairment of recognition memory in mice repeatedly treated with methamphetamine: involvement of allosteric potentiation of nicotinic acetylcholine receptors and dopaminergic-ERK1/2 systems.
Galantamine, a drug used to treat Alzheimer's disease, inhibits acetylcholinesterase (AChE) and allosterically modulates nicotinic acetylcholine receptors (nAChRs) resulting in stimulation of catecholamine neurotransmission. In this study, we investigated whether galantamine exerts cognitive-improving effects through the allosteric modulation of nAChRs in an animal model of methamphetamine (Meth) psychosis. The mice treated with Meth (1 mg/kg.d) for 7 d showed memory impairment in a novel object recognition test. Galantamine (3 mg/kg) ameliorated the memory impairment, and it increased the extracellular dopamine release in the prefrontal cortex (PFC) of Meth-treated mice. Donepezil, an AChE inhibitor (1 mg/kg) increased the extracellular ACh release in the PFC, whereas it had no effect on the memory impairment in Meth-treated mice. The nAChR antagonist, mecamylamine, and dopamine D1 receptor antagonist, SCH 23390, blocked the ameliorating effect of galantamine on Meth-induced memory impairment, whereas the muscarinic AChR antagonist, scopolamine, had no effect. The effects of galantamine on extracellular dopamine release were also antagonized by mecamylamine. Galantamine attenuated the defect of the novelty-induced activation of extracellular signal-regulated kinase 1/2 (ERK1/2). The ameliorating effect of galantamine on recognition memory in Meth-treated mice was negated by microinjection of an ERK inhibitor, PD98059, into the PFC. These results suggest that the ameliorating effect of galantamine on Meth-induced memory impairment is associated with indirect activation of dopamine D1 receptor-ERK1/2 following augmentation with dopaminergic neurotransmission in the PFC through the allosteric activation of nAChRs. Galantamine could be a useful therapeutic agent for treating cognitive deficits in schizophrenia/Meth psychosis, as well as Alzheimer's disease.


Anti-MAdCAM-1 antibody (PF-00547659) for active refractory Crohn's disease in Japanese and Korean patients: the OPERA study.
PF-00547659 is a monoclonal antibody against human mucosal addressin cell adhesion molecule-1 (MAdCAM-1) that prevents the binding of α4β7+ lymphocytes to MAdCAM-expressing sites in the gastrointestinal tract with high affinity and selectivity, and is being developed for the treatment of Crohn's disease (CD). OPERA is a randomized, multicenter, double-blind, placebo-controlled study to investigate the efficacy, safety, and pharmacokinetics of PF-00547659 following subcutaneous administration in subjects with active CD, a history of failure or intolerance to anti-tumor necrosis factor and/or immunosuppressants, high-sensitivity C-reactive protein > 3.0 mg/L, and ulcers on colonoscopy. The primary endpoint was Crohn's Disease Activity Index-70 response at week 8 or 12. Subpopulation analyses for Asian subjects were performed as some differences are observed in genetics and clinical phenotypes in Asian CD patients compared with Western patients. In this study, 265 CD subjects were randomized, with a subpopulation of 21 subjects (8 Japanese and 13 Korean) defined as the Asian population. In the overall and Asian populations; PF-00547659 was pharmacologically active as evidenced by soluble MAdCAM and circulating β7+ central memory CD4+ T-lymphocytes, although no clear evidence of efficacy was observed in any clinical endpoints; pharmacokinetics of PF-00547659 in the Asian subpopulation was generally comparable to the overall population; and the safety profile of PF-00547659 appeared acceptable up to 12 weeks of treatment. In the overall and Asian populations, efficacy of PF-00547659 could not be demonstrated using any clinical endpoints compared with placebo. Pharmacokinetics and safety of PF-00547659 were generally comparable. Further studies with larger numbers of patients are required to confirm our results. (Trial Registration Number: NCT01276509).


Brain cancer incidence, mortality and case survival: observations from two Australian cancer registries.
Data from 2 Australian cancer registries covering a population of 1.7 million people were combined for the purposes of analysing brain cancer incidence, mortality and survival patterns for the time period 1978 through 1992. A total of 1,752 cases of primary brain cancer were registered, representing age-standardised incidence rates of 6.7 per 100,000 in men and 4.6 in women. Histological confirmation was available for 94% of cases. The incidence rate among persons aged 75 or over was higher during 1986-1992 than during 1978-1985, the rate for men increasing from 16.3 to 26.2 and that for women increasing from 9.7 to 18.0. The largest increases in this age group occurred for cases of glioblastoma multiforme. During the study period, 1,411 brain cancer deaths were notified to the 2 registries at age-standardised rates of 5.3 in men and 3.4 in women. Mortality rates among persons aged 75 years or older were higher during 1986-1992 than 1978-1985, increasing from 15.7 to 28.4 in men and from 10.1 to 15.3 in women. Only among men aged 15-49 years was a decline in mortality rates observed, from 3.3 to 2.4. Survival analyses indicated that age and histological type were the most powerful prognostic indicators. There was no improvement in 5-year survival for any of the age groups or histological types. An improvement in 36-month survival was noted for the 15-49 year age group diagnosed with gliomas other than glioblastoma multiforme.


Ellagic acid inhibits lipopolysaccharide-induced expression of enzymes involved in the synthesis of prostaglandin E2 in human monocytes.
Ellagic acid, a natural polyphenol found in certain fruits, nuts and vegetables, has in recent years been the subject of intense research within the fields of cancer and inflammation. Pain, fever and swelling, all typical symptoms of inflammation, are ascribed to elevated levels of PGE2. In the present study, we have investigated the effects of ellagic acid on PGE2 release and on prostaglandin-synthesising enzymes in human monocytes. Ellagic acid was found to inhibit Ca ionophore A23187-, phorbol myristate acetate- and opsonised zymosan-induced release of PGE2 from monocytes pre-treated with the inflammatory agent lipopolysaccharide. Ellagic acid suppressed the lipopolysaccharide-induced increase in protein expression of cyclo-oxygenase-2 (COX-2), microsomal PGE synthase-1 (mPGEs-1) and cytosolic phospholipase A2alpha (cPLA2alpha), while it had no effect on the constitutively expressed COX-1 protein. Ellagic acid had no apparent inhibitory effect on these enzymes when the activities were determined in cell-free assays. We conclude that the inhibitory effect of ellagic acid on PGE2 release from monocytes is due to a suppressed expression of COX-2, mPGEs-1 and cPLA2alpha, rather than a direct effect on the activities of these enzymes.


Membrane fusion in a model system. Mucocyst secretion in Tetrahymena.
The freeze-fracture, freeze-etch technique can be employed to reveal new details of the process of fusion of two unit membranes For this study, mucocyst discharge in Tetrahymena pyriformis provides a model system with certain general implications The undischarged mature mucocyst is a saclike, membrane-bound, secretory vesicle containing crystalline material The organelle tip finds its way toward a special site, a rosette of 150 A diameter particles within the plasma membrane. To match this site, the mucocyst membrane forms an annulus of 110 A diameter particles, above whose inner edge the rosette particles sit. Discharge of some mucocysts is triggered by fixation. As discharge proceeds, the organelle becomes spherical and its content changes from crystalline to amorphous. The cytoplasm between the two matching membrane sites is squeezed away and the membranes fuse Steps in membrane reorganization can be reconstructed from changes in rosette appearance in the fracture faces. First, a depression in the rosette-the fusion pocket-forms. The rosette particles spread at the lip as the pocket deepens and enlarges from 60 to 200 nm. The annulus particles then become visible at the lip, indicating completed fusion of the A fracture faces of mucocyst and plasma membranes The remaining B faces of the two membranes have opposite polarities When the content of the mucocyst is released, the edges of these faces join so that the unit membrane runs uninterruptedly around the lip and into the pocket.


Polyp volume versus linear size measurements at CT colonography: implications for noninvasive surveillance of unresected colorectal lesions.
Proposed surveillance of unresected medium-sized (6.0-9.9 mm) polyps will require reliable detection of small incremental changes. The purpose of this study was to assess the reproducibility of linear versus volume measurements of polyp size at CT colonography (CTC) and to correlate results with a hemispheric model. The study group consisted of 30 polyps on supine and prone CTC data sets. Measurements were performed separately by two experienced radiologists using the same CTC system (Vitrea 2), resulting in 120 linear and volume measurements. Linear size was defined as the longest dimension among the 2D multiplanar reconstruction views. Semiautomated volume determination required manual tracing of polyp boundaries on 2D multiplanar reconstruction views. The relative error between reviewers 1 and 2 was defined as 100 x (/D1 - D2/)/D(ave) for linear size (D) and as 100 x (/V1 - V2)/V(ave) for volume (V), where ave is average. The mean relative error for linear size and volume measurement was 10.4% +/- 10.7% and 16.9% +/- 13.2%, respectively. Median linear size and volume of polyps were 9.4 mm and 270 mm3, respectively. CTC-derived volumes for medium-sized polyps closely approximated hemispheric volume (V = (pi/12) x D(b), where b =3.13, r2 = 0.90). Small incremental changes in hemispheric size result in a 3:1 relative change in volume versus diameter, such that a 1-mm diameter change in a medium-sized hemispheric polyp results in a relative change in linear size and volume ranging from approximately 11-18% and 31-53%, respectively. Because changes in polyp volume are amplified compared with linear dimension, volume measurement rather than diameter measurement will better allow detection of small incremental changes in polyp size using CTC.


Assessment of emerging polar organic pollutants linked to contaminant pathways within an urban estuary using non-targeted analysis.
A comprehensive, non-targeted analysis of polar organic pollutants using high resolution/accurate mass (HR/AM) mass spectrometry approaches has been applied to water samples from San Francisco (SF) Bay, a major urban estuary on the western coast of the United States, to assess occurrence of emerging contaminants and inform future monitoring and management activities. Polar Organic Chemical Integrative Samplers (POCIS) were deployed selectively to evaluate the influence of three contaminant pathways: urban stormwater runoff (San Leandro Bay), wastewater effluent (Coyote Creek, Lower South Bay), and agricultural runoff (Napa River). Grab samples were collected before and after deployment of the passive samplers to provide a quantitative snapshot of contaminants for comparison. Composite samples of wastewater effluent (24 hours) were also collected from several wastewater dischargers. Samples were analyzed using liquid-chromatography coupled to high resolution mass spectrometry. Resulting data were analyzed using a customized workflow designed for high-fidelity detection, prioritization, identification, and semi-quantitation of detected molecular features. Approximately 6350 compounds were detected in the combined data set, with 424 of those compounds tentatively identified through high quality spectral library match scores. Compounds identified included ethoxylated surfactants, pesticide and pharmaceutical transformation products, polymer additives, and rubber vulcanization agents. Compounds identified in samples were reflective of the apparent sources and pathways of organic pollutant inputs, with stormwater-influenced samples dominated by additive chemicals likely derived from plastics and vehicle tires, as well as ethoxylated surfactants.


Survival in second degree atrioventricular block.
Two hundred and 14 patients with chronic second degree heart block were seen and followed up in the Devon Heart Block and Bradycardia Survey between 1968 and 1982. The patients were divided into three groups according to the type of block. In group 1 there were 77 patients with Mobitz type I block (mean age 69 years), in group 2, 86 patients with Mobitz type II block (mean age 74 years), and in group 3, 51 with 2:1 or 3:1 block (mean age 75 years). The five year survival was similar in all groups, being 57%, 61%, and 53% in groups 1, 2, and 3 respectively. The presence or absence of bundle branch block did not appear to influence prognosis. In particular, patients in group 1 without bundle branch block did not fare any better than those in group 2 both with and without bundle branch block. One hundred and three of the patients were fitted with pacemakers, the proportion being greatest in group 2. In each group a significantly larger number of paced patients survived than unpaced. The five year survival for all the paced patients in the study was 78% compared with 41% for the unpaced. Since the paced patients were slightly younger than the unpaced two age matched groups of 74 patients each were selected from the paced and unpaced patients, but the five year survival of those paced was still significantly better. It is concluded that in the patients in the present study chronic Mobitz type I block has a similar prognosis to that of Mobitz type II block. Unpaced patients with both types did very badly, whereas those fitted with pacemakers had a five year survival similar to that expected for the normal population. These results refute the benign reputation of chronic Mobitz type I block and imply that patients with this condition should be considered for pacemaker implantation on similar criteria to those adopted for patients with higher degrees of block.


Differential var gene expression in the organs of patients dying of falciparum malaria.
Sequestration of parasitized erythrocytes in the microcirculation of tissues is thought to be important in the pathogenesis of severe falciparum malaria. A major variant surface antigen, var/Plasmodium falciparum erythrocyte membrane protein 1, expressed on the surface of the infected erythrocyte, mediates cytoadherence to vascular endothelium. To address the question of tissue-specific accumulation of variant types, we used the unique resource generated by the clinicopathological study of fatal paediatric malaria in Blantyre, Malawi, to analyse var gene transcription in patients dying with falciparum malaria. Despite up to 102 different var genes being expressed by P. falciparum populations in a single host, only one to two of these genes were expressed at high levels in the brains and hearts of these patients. These major var types differed between organs. However, identical var types were expressed in the brains of multiple patients from a single malaria season. These results provide the first evidence of organ-specific accumulation of P. falciparum variant types and suggest that parasitized erythrocytes can exhibit preferential binding in the body, supporting the hypothesis of cytoadherence-linked pathogenesis.


Juvenile delinquency and attention deficit disorder: boys' developmental trajectories from age 3 to age 15.
This article describes a longitudinal analysis of the behavior of a birth cohort of 435 boys. 4 groups were defined at age 13 on the basis of both self-reported delinquent behavior and professional diagnosis of Attention Deficit Disorder: ADD + delinquent, ADD only, delinquent only, and nondisordered. Biennial correlates of delinquency (antisocial behavior problems, verbal intelligence, reading difficulty, and family adversity) were traced across childhood. The ADD + delinquent boys consistently fared the worst on the assessments of family adversity, verbal intelligence, and reading. Their antisocial behavior began before school age, escalated at school entry, and persisted into adolescence. The ADD-only boys had normal family, intelligence, and reading scores, and showed only mild antisocial behavior in middle childhood. The delinquent-only boys showed no early risk from family, low intelligence, or reading deficit, and remained relatively free of conduct problems until they initiated delinquency at age 13. Persistence of criminal offending beyond adolescence is predicted for the ADD + delinquent boys.


Hypermethylation of CD19 promoter enables antigen-negative escape to CART-19 in vivo and in vitro.
Anti-CD19 chimeric antigen receptor T cells (CART-19) frequently induce remissions in hemato-oncological patients with recurred and/or refractory B-cell tumors. However, malignant cells sometimes escape the immunotherapeutic targeting by CD19 gene mutations, alternative splicing or lineage switch, commonly causing lack of CD19 expression on the surface of neoplastic cells. We assumed that, in addition to the known mechanisms, other means could act on CD19 to drive antigen-negative relapse. Herein, we studied the mechanism of antigen loss in an in vivo CD19-negative recurrence model of chronic lymphocytic leukemia (CLL) to CART-19, established using NOD-scid IL2Rgnull mice and HG3 cell line. We validated our findings in vitro in immortalized B-cell lines and primary CLL cells. In our in vivo CLL recurrence model, up to 70% of CART-19-treated mice eventually recurred with CD19-negative disease weeks after initial positive response. We found that the lack of CD19 expression was caused by promoter DNA hypermethylation. Importantly, the expression loss was partially reversible by treatment with a demethylating agent. Moreover, this escape mechanism was common for 3 B-cell immortalized lines as well as primary CLL cells, as assessed by in vitro coculture experiments. Epigenetically driven antigen escape could represent a novel, yet at least partially reversible, means of CD19 loss to CART-19 in B-cell tumors.


Abdominal aortic aneurysm in women: prevalence, risk factors, and implications for screening.
Accurate data regarding the prevalence and associated risk factors for aneurysmal disease is essential when determining the appropriateness of screening for abdominal aortic aneurysms (AAA). Although women are poorly represented in most large studies of AAA prevalence, the US Preventative Services Task Force recently recommended against primary screening for AAA in women. The purpose of this analysis was to define the prevalence and risk factors associated with the development of AAA in women. A free duplex ultrasound screening was offered to men and women with cardiovascular risk factors or a family history of AAA. Patients were recruited through advertising at local screening centers and screenings were performed between 2004 and 2006. Demographic information and cardiovascular and aneurysmal disease risk factors were obtained for each patient through a questionnaire. A total of 17,540 subjects were screened for AAA, including 10,012 women (mean age 69.6 years) and 7528 men (mean age 70.0 years). Univariate and multivariable logistic regression analysis was performed on the subset of women that were screened to determine risk factors for and prevalence of AAA. Seventy-four aneurysms were detected in women (including four aneurysms >5 cm diameter and 70 aneurysms 3 to 5 cm diameter) while 291 were detected in men, resulting in prevalence rates of 0.7% and 3.9%, respectively. Increasing age (odds ratio [OR]= 4.57, 95% confidence interval [CI] 1.98 to 10.54, P < .0001), history of tobacco use (OR = 3.29, 95% CI 1.86 to 5.80, P < .0001), and cardiovascular disease (OR= 3.57, 95% CI 2.19 to 5.84, P < .0001) were independently associated with AAA in women on univariate and multivariable analysis. Women with multiple atherosclerotic risk factors were more commonly found to have AAAs and had a prevalence rate of AAA as high as 6.4%. Although the medical literature suggests a low prevalence rate of AAA in women in the general population, specific risk factors are associated with the development of AAA, and subgroups of women can be identified that are at a substantially increased risk of aneurysmal disease. In particular, elevated rates of AAA were found among women of advanced age (> or =65 years) with a history of smoking or heart disease. These data support the notion that women with such risk factors should be considered for AAA screening.


Color manipulation through microchip tinting for colorimetric detection using hue image analysis.
Colorimetry with microfluidic devices has been proven to be an advantageous method for in situ analyses where limited resources and rapid response for untrained users are desired. Image analysis using a small camera or cell phone can be easily incorporated for an objective readout, eliminating variations from normal differences in color perception and environmental factors during analysis. The image analysis using the parameter hue, for example, has been utilized as a highly effective, objective analysis method that correlates with the psychological way color is perceived. Hue analysis, however, is best used for colorimetric reactions that result in distinct changes from one color to a markedly different color and can be inadequate to distinguish between subtle or monotonal (colorless-to-colored) color changes. We address this with three unique color manipulation (i.e., tinting) techniques that provide greater discrimination with such color changes, thus yielding improved limits of detection for various colorimetric reactions that may have previously been limited. Tinting is invoked through dyeing the reagent substrate, colored printing the device, or colored lighting during image capture, and is shown to effectively shift the background color of the reaction detection area. Hydrogen peroxide, a constituent of peroxide-based explosives, is associated with a monochromatic color change upon reaction, and this is used to demonstrate the effectiveness of the tinting methods in improving the limit of detection from an undetectable color change to 0.1 mg mL-1.


Sustaining consistent condom use among female sex workers by addressing their vulnerabilities and strengthening community-led organizations in India.
Between 2014 and 2017, a program aimed at reducing HIV risk and promoting safe sex through consistent use of condoms sought to work through addressing social and economic vulnerabilities and strengthening community-led organizations (COs) of female sex workers (FSWs). This study examines if the program was effective by studying relationship between strengthening of COs, vulnerability reduction, and sustaining of consistent condom use behavior among FSWs. We used a longitudinal study design to assess the change in outcomes. A three-stage sampling design was used to select FSWs for the study. Panel data of 2085 FSWs selected from 38 COs across five states of India was used to examine the change in various outcomes from 2015 (Survey Round 1) to 2017 (Survey Round 2). The CO level program pillar measuring institutional development assessed performance of COs in six domains critical for any organization's functionality and sustainability: governance, project management, financial management, program monitoring, advocacy and networking, and resource mobilization. Overall, 32 indicators from all these domains were used to compute the CO strength score. A score was computed by taking mean of average dimension scores. The overall score was divided into two groups based on the median cutoff; COs which scored below the median were considered to have low CO strength, while COs which scored above or equal to median were considered to have high CO strength. Multivariable regression modeling techniques were used to examine the effect of program pillars on outcome measures. Analyses showed a significant improvement in the strength of the COs over time; percentage of COs having high strength improved from 50% in 2015 to 87% in Round 2. The improvement in CO's strength increased financial security (Adjusted Odds Ratio [AOR]: 2.18, p<0.01), social welfare security (AOR: 1.71, p<0.01), and socio-legal security (AOR: 2.20, p<0.01) among FSWs. Further, improvement in financial security led to significant increase in consistent condom use with client among FSWs (AOR: 1.69, p<0.01) who were members of COs having high strength. Sustained consistent condom use was positively associated with young age (<30 years), ability to negotiate with clients for condom use, membership in self-help groups, high self-efficacy, self-confidence, and client solicitation in streets and brothels. Improving financial security and strengthening FSW led CO can improve sustained and consistent condom use. In addition, the program should focus on enhancing ability of FSWs to negotiate with clients for condom use, promote membership in self-help groups and target FSWs who are 30 years or older, and soliciting from homes to sustain consistent condom use across all FSWs.


CD20 therapies in multiple sclerosis and experimental autoimmune encephalomyelitis - Targeting T or B cells?
MS is widely considered to be a T cell-mediated disease although T cell immunotherapy has consistently failed, demonstrating distinct differences with experimental autoimmune encephalomyelitis (EAE), an animal model of MS in which T cell therapies are effective. Accumulating evidence has highlighted that B cells also play key role in MS pathogenesis. The high frequency of oligoclonal antibodies in the CSF, the localization of immunoglobulin in brain lesions and pathogenicity of antibodies originally pointed to the pathogenic role of B cells as autoantibody producing plasma cells. However, emerging evidence reveal that B cells also act as antigen presenting cells, T cell activators and cytokine producers suggesting that the strong efficacy of anti-CD20 antibody therapy observed in people with MS may reduce disease progression by several different mechanisms. Here we review the evidence and mechanisms by which B cells contribute to disease in MS compared to findings in the EAE model.


Family meals and body mass index among adolescents: effects of gender.
Family meals have been identified as a protective factor against obesity among youth. However, gender specificities with respect to the relationship between the frequency of family meals and body mass index (BMI) have not been investigated. The purpose of this study was to examine the relationship between the frequency of family meals and BMI in male and female adolescents, while controlling for potential confounding factors associated with BMI, such as parental education, adolescent's age, and snack-food eating. Research participants were 734 male and 1030 female students (mean age, 14.12 years, SD = 1.62) recruited from middle schools and high schools in the capital region of Canada. Participants completed validated, self-report measures to assess the frequency of family meals and the risk factors associated with increased BMI, which was derived from objective measures of height and weight. After controlling for proposed confounding variables, a higher frequency of family meals was associated with lower BMI in females, but not in males. A Z-transformation test of the homogeneity of adjusted correlation coefficients showed a significant trend (p = 0.06), indicating that the relationship between family meals and BMI is stronger in females than males, consistent with our regression analyses. Our findings suggest that eating together as a family may be a protective factor against obesity in female adolescents, but not in male adolescents. Findings from this study have important implications for parents and health care practitioners advocating for more frequent family meals as part of a comprehensive obesity prevention and treatment program for female adolescents.


Image analysis-derived metrics of histomorphological complexity predicts prognosis and treatment response in stage II-III colon cancer.
The complexity of tumor histomorphology reflects underlying tumor biology impacting on natural course and response to treatment. This study presents a method of computer-aided analysis of tissue sections, relying on multifractal (MF) analyses, of cytokeratin-stained tumor sections which quantitatively evaluates of the morphological complexity of the tumor-stroma interface. This approach was applied to colon cancer collection, from an adjuvant treatment randomized study. Metrics obtained with the method acted as independent markers for natural course of the disease, and for benefit of adjuvant treatment. Comparative analyses demonstrated that MF metrics out-performed standard histomorphological features such as tumor grade, budding and configuration of invasive front. Notably, the MF analyses-derived "αmax" -metric constitutes the first response-predictive biomarker in stage II-III colon cancer showing significant interactions with treatment in analyses using a randomized trial-derived study population. Based on these results the method appears as an attractive and easy-to-implement tool for biomarker identification.


The C-terminal region of a Lys49 myotoxin mediates Ca2+ influx in C2C12 myotubes.
Myotoxins are abundant components of snake venoms, being a significant public health problem worldwide. Among them, Lys49 phospholipase A(2) homologue myotoxins cause extensive necrosis in skeletal muscle tissue. Their mechanisms of action are still poorly understood, but there is evidence that the C-terminal region is involved in membrane damage leading to myotoxicity. To investigate the effect of the C-terminal peptide 115-129 of Agkistrodon contortrix laticinctus myotoxin on the plasma membrane of myoblasts and myotubes, the entry of Ca(2+) was monitored by fluorescence imaging, and the ensuing cytotoxicity was determined. The myotoxin synthetic peptide was found to act selectively on myotubes, which were rapidly overloaded with Ca(2+) with ensuing necrosis. The profile of intracellular Ca(2+) increase induced by the C-terminal peptide, but not by its scrambled version control, reproduces the second, prominent wave of the biphasic response documented in previous studies using whole Lys49 myotoxins. These observations provide relevant insights into the mechanism of action of this family of toxins, with implications for the understanding of their structure-function relationships.


The single and joint toxicity effects of chlorpyrifos and beta-cypermethrin in zebrafish (Danio rerio) early life stages.
Water environment pollution caused by the widespread application of beta-cypermethrin (BCP) and chlorpyrifos (CPF) in agriculture has attracted extensive concern of the world. In this study, zebrafish was used as a model to investigate the individual and joint toxicity of BCP and CPF. In the acute toxicity test, 3 hpf embryos were exposed to various concentrations of CPF, BCP and their binary mixtures (MIX) for 96h. The results indicated that these two pesticides and mixtures induced malformation and death in larvae, and affected hatchability. These two pesticides in mixtures were verified to act together in a synergistic manner under experimental conditions. Oxidative stress assaying manifested that CPF, BCP and MIX altered CAT, SOD and GST activities and MDA content, resulting in oxidative damage in larvae. By pathology analysis, CPF (236μg/L), BCP (5.9μg/L) and MIX (236μg/L CPF+5.9μg/L BCP) were found to trigger liver lesions and promote apoptosis in tissues. The transcriptome sequencing suggested that ECM- receptor interaction, focal adhesion, cell cycle, DNA replication, phototransduction and adherens junction pathways were closely associated with the toxicity of these two pesticides.


Characterising demographics, knowledge, practices and clinical care among patients attending sickle cell disease clinics in Eastern Uganda.
Background: In Uganda to date, there are neither established registries nor descriptions of facility-based sickle cell disease (SCD) patient characteristics beyond the central region. Here, we summarize data on the baseline clinical characteristics and routine care available to patients at four clinics in Eastern Uganda as a prelude to a clinical trial. Methods: Between February and August 2018, we conducted a cross-sectional survey of patients attending four SCD clinics in Mbale, Soroti, Atutur and Ngora, all in Eastern Uganda, the planned sites for an upcoming clinical trial (H-PRIME: ISRCTN15724013). Data on socio-demographic characteristics, diagnostic methods, clinic schedules, the use of prophylactic and therapeutic drugs, clinical complications and patient understanding of SCD were collected using a structured questionnaire. Results: Data were collected on 1829 patients. Their ages ranged from 0 to 64 years with a median (IQR) of 6 (3-11) years. 50.9% of participants were male. The majority (1151; 62.9%) reported a positive family history for SCD. Approximately half knew that SCD is inherited from both parents but a substantial proportion did not know how SCD is transmitted and small numbers believed that it is acquired by either transfusion or from other people. Only 118/1819 (6.5%) participants had heard about or were using hydroxyurea while 356/1794 (19.8%) reported stigmatization. Participants reported a median of three (IQR 1-4) hospital admissions during the preceding 12 months; 80.8% had been admitted at least once, while 14.2% had been admitted more than five times. Pain was the most common symptom, while 83.9% of those admitted had received at least one blood transfusion. Conclusion: The majority of patients attending SCD clinics in Eastern Uganda are children and few are currently being treated with hydroxyurea. The data collected through this facility-based survey will provide background data that will be useful in planning for the H-PRIME trial.


Estimation of Achievable Oxygen Consumption Following Transfusion With Rejuvenated Red Blood Cells.
Erythrocyte storage induces a nonphysiological increase in hemoglobin-oxygen affinity (quantified by low p50, the oxygen tension at 50% hemoglobin saturation), which can be restored through biochemical rejuvenation. The objective was to mathematically model the impact of transfusing up to 3 standard allogeneic units or rejuvenated units on oxygen delivery (DO2) and oxygen consumption (VO2). Oxygen dissociation curves were generated from additive solution-1 red blood cell (RBC) leukoreduced units (n = 7) before and after rejuvenation following manufacturer's instructions. Two of these units were used to prepare standard or rejuvenated donor RBC and added to samples of fresh whole blood. These admixtures were used to construct an in vitro transfusion model of postoperative anemia and determine a linear equation for calculating the sample p50, which was subsequently used to calculate DO2 and VO2 after simulated transfusions. Whole blood-packed red blood cell unit admixture p50s could be predicted from a linear model including the p50 of its components, the mass fraction of the transfused component, and interaction terms (R2 = .99, P < 0.001). Transfusion with standard units slightly, but significantly, increased projected DO2 compared with rejuvenated units (P = 0.03), but rejuvenated units markedly increased projected VO2 (P = 0.03). Standard units did not significantly change VO2 relative to pre-transfusion levels (P > 0.1). Using high-p50, rejuvenated RBC in simulated transfusions greatly improved projected VO2, indicating the potential for increased end-organ oxygen availability compared with standard transfusion. Patient capacity to increase cardiac output after cardiac surgery may be limited. Transfusing high-p50 RBC in this setting may improve the perioperative care of these patients.


Immunotherapy and gene therapy of cancer.
In the past decade, immunotherapies have been developed that are capable of causing prolonged cancer regressions in selected patients with advanced metastatic disease. In the past year, attempts at the gene therapy of cancer have begun. These experimental cancer treatments deserve vigorous exploration.


First-line treatment selection and early monitoring patterns in chronic phase-chronic myeloid leukemia in routine clinical practice: SIMPLICITY.
Achieving successful outcomes in chronic phase-chronic myeloid leukemia (CP-CML) requires careful monitoring of cytogenetic/molecular responses (CyR/MR). SIMPLICITY (NCT01244750) is an observational study exploring tyrosine kinase inhibitor use and management patterns in patients with CP-CML receiving first-line imatinib (n = 416), dasatinib (n = 418) or nilotinib (n = 408) in the US and 6 European countries in routine clinical practice. Twelve-month follow-up data of 1242 prospective patients (enrolled October 01 2010-September 02 2015) are reported. 81% of patients had baseline comorbidities. Treatment selection was based on perceived efficacy over patient comorbidity profile. There was a predominance of imatinib-treated patients enrolled earlier in the study, with subsequent shift toward dasatinib- and nilotinib-treated patients by 2013/2014. Monitoring for either CyR/MR improved over time and was documented for 36%, 82%, and 95% of patients by 3, 6, and 12 months, respectively; 5% had no documentation of CyR/MR monitoring during the first year of therapy. Documentation of MR/CyR testing was higher in Europe than the US (P < .001) and at academic versus community practices (P = .001). Age <65 years, patients being followed at sites within Europe, those followed at academic centers and patients no longer on first-line therapy were more likely to be monitored by 12 months. SIMPLICITY demonstrates that the NCCN and ELN recommendations on response monitoring have not been consistently translated into routine clinical practice. In the absence of appropriate monitoring practices, clinical response to TKI therapy cannot be established, any needed changes to treatment strategy will thus not be implemented, and long-term patient outcomes are likely to be impacted.


Orthogonal polarization fiber gyroscope with increased stability and resolution.
The orthogonal polarization fiber gyroscope (OPFG) is an interferometric fiber gyroscope design that requires no phase bias in the fiber ring and is insensitive to light-source intensity noise. However, in the original OPFG [Hitachi Rev. 33, 215 (1984)], environmental changes caused first-order false rotation signals. We propose and experimentally verify modifications that eliminate the first-order sensitivity to environmental changes and that improve the gyroscope's resolution as well. We believe that this modified OPFG is the first interferometric fiber gyroscope capable of stable, high-sensitivity measurement that contains only reciprocal optical elements.


A Transcriptome-Wide Association Study Identifies Novel Candidate Susceptibility Genes for Pancreatic Cancer.
Although 20 pancreatic cancer susceptibility loci have been identified through genome-wide association studies in individuals of European ancestry, much of its heritability remains unexplained and the genes responsible largely unknown. To discover novel pancreatic cancer risk loci and possible causal genes, we performed a pancreatic cancer transcriptome-wide association study in Europeans using three approaches: FUSION, MetaXcan, and Summary-MulTiXcan. We integrated genome-wide association studies summary statistics from 9040 pancreatic cancer cases and 12 496 controls, with gene expression prediction models built using transcriptome data from histologically normal pancreatic tissue samples (NCI Laboratory of Translational Genomics [n = 95] and Genotype-Tissue Expression v7 [n = 174] datasets) and data from 48 different tissues (Genotype-Tissue Expression v7, n = 74-421 samples). We identified 25 genes whose genetically predicted expression was statistically significantly associated with pancreatic cancer risk (false discovery rate < .05), including 14 candidate genes at 11 novel loci (1p36.12: CELA3B; 9q31.1: SMC2, SMC2-AS1; 10q23.31: RP11-80H5.9; 12q13.13: SMUG1; 14q32.33: BTBD6; 15q23: HEXA; 15q26.1: RCCD1; 17q12: PNMT, CDK12, PGAP3; 17q22: SUPT4H1; 18q11.22: RP11-888D10.3; and 19p13.11: PGPEP1) and 11 at six known risk loci (5p15.33: TERT, CLPTM1L, ZDHHC11B; 7p14.1: INHBA; 9q34.2: ABO; 13q12.2: PDX1; 13q22.1: KLF5; and 16q23.1: WDR59, CFDP1, BCAR1, TMEM170A). The association for 12 of these genes (CELA3B, SMC2, and PNMT at novel risk loci and TERT, CLPTM1L, INHBA, ABO, PDX1, KLF5, WDR59, CFDP1, and BCAR1 at known loci) remained statistically significant after Bonferroni correction. By integrating gene expression and genotype data, we identified novel pancreatic cancer risk loci and candidate functional genes that warrant further investigation.


Pay for performance, quality of care, and outcomes in acute myocardial infarction.
Pay for performance has been promoted as a tool for improving quality of care. In 2003, the Centers for Medicare & Medicaid Services (CMS) launched the largest pay-for-performance pilot project to date in the United States, including indicators for acute myocardial infarction. To determine if pay for performance was associated with either improved processes of care and outcomes or unintended consequences for acute myocardial infarction at hospitals participating in the CMS pilot project. An observational, patient-level analysis of 105,383 patients with acute non-ST-segment elevation myocardial infarction enrolled in the Can Rapid Risk Stratification of Unstable Angina Patients Suppress Adverse Outcomes With Early Implementation of the American College of Cardiology/American Heart Association (ACC/AHA) Guidelines (CRUSADE) national quality-improvement initiative. Patients were treated between July 1, 2003, and June 30, 2006, at 54 hospitals in the CMS program and 446 control hospitals. The differences in the use of ACC/AHA class I guideline recommended therapies and in-hospital mortality between pay for performance and control hospitals. Among treatments subject to financial incentives, there was a slightly higher rate of improvement for 2 of 6 targeted therapies at pay-for-performance vs control hospitals (odds ratio [OR] comparing adherence scores from 2003 through 2006 at half-year intervals for aspirin at discharge, 1.31; 95% confidence interval [CI], 1.18-1.46 vs OR, 1.17; 95% CI, 1.12-1.21; P = .04) and for smoking cessation counseling (OR, 1.50; 95% CI, 1.29-1.73 vs OR, 1.28; 95% CI, 1.22-1.35; P = .05). There was no significant difference in a composite measure of the 6 CMS rewarded therapies between the 2 hospital groups (change in odds per half-year period of receiving CMS therapies: OR, 1.23; 95% CI, 1.15-1.30 vs OR, 1.17; 95% CI, 1.14-1.20; P = .16). For composite measures of acute myocardial infarction treatments not subject to incentives, rates of improvement were not significantly different (OR, 1.09; 95% CI, 1.05-1.14 vs OR, 1.08; 95% CI, 1.06-1.09; P = .49). Overall, there was no evidence that improvements in in-hospital mortality were incrementally greater at pay-for-performance sites (change in odds of in-hospital death per half-year period, 0.91; 95% CI, 0.84-0.99 vs 0.97; 95% CI, 0.94-0.99; P = .21). Among hospitals participating in a voluntary quality-improvement initiative, the pay-for-performance program was not associated with a significant incremental improvement in quality of care or outcomes for acute myocardial infarction. Conversely, we did not find evidence that pay for performance had an adverse association with improvement in processes of care that were not subject to financial incentives. Additional studies of pay for performance are needed to determine its optimal role in quality-improvement initiatives.


Economic Evaluation alongside Multinational Studies: A Systematic Review of Empirical Studies.
This study seeks to explore methods for conducting economic evaluations alongside multinational trials by conducting a systematic review of the methods used in practice and the challenges that are typically faced by the researchers who conducted the economic evaluations. A review was conducted for the period 2002 to 2012, with potentially relevant articles identified by searching the Medline, Embase and NHS EED databases. Studies were included if they were full economic evaluations conducted alongside a multinational trial. A total of 44 studies out of a possible 2667 met the inclusion criteria. Methods used for the analyses varied between studies, indicating a lack of consensus on how economic evaluation alongside multinational studies should be carried out. The most common challenge appeared to be related to addressing differences between countries, which potentially hinders the generalisability and transferability of results. Other challenges reported included inadequate sample sizes and choosing cost-effectiveness thresholds. It is recommended that additional guidelines be developed to aid researchers in this area and that these be based on an understanding of the challenges associated with multinational trials and the strengths and limitations of alternative approaches. Guidelines should focus on ensuring that results will aid decision makers in their individual countries.


Diamond formation due to a pH drop during fluid-rock interactions.
Diamond formation has typically been attributed to redox reactions during precipitation from fluids or magmas. Either the oxidation of methane or the reduction of carbon dioxide has been suggested, based on simplistic models of deep fluids consisting of mixtures of dissolved neutral gas molecules without consideration of aqueous ions. The role of pH changes associated with water-silicate rock interactions during diamond formation is unknown. Here we show that diamonds could form due to a drop in pH during water-rock interactions. We use a recent theoretical model of deep fluids that includes ions, to show that fluid can react irreversibly with eclogite at 900 °C and 5.0 GPa, generating diamond and secondary minerals due to a decrease in pH at almost constant oxygen fugacity. Overall, our results constitute a new quantitative theory of diamond formation as a consequence of the reaction of deep fluids with the rock types that they encounter during migration. Diamond can form in the deep Earth during water-rock interactions without changes in oxidation state.


Identification of sas, a conserved gene cluster involved in the regulation of aerial mycelium formation in Streptomyces griseus.
We cloned a DNA fragment that suppressed the aerial-mycelium-deficient phenotype in an amfS mutant of Streptomyces griseus when it was introduced into the cells via a high-copy-number plasmid. The sasABCDR gene cluster was identified as being responsible for this suppressive activity. The proteins encoded by sasABCD were of unknown function, but the operon structure was found to be conserved in all the strains of Streptomyces spp. and related organisms whose genomes have been sequenced. sasR, the flanking opposite coding sequence, encoded a putative DNA-binding protein. Subcloning revealed that the presence of all five coding sequences was essential for complete suppression. Scanning electron microscopy of Streptomyces griseus strains carrying the sas gene cluster at a high copy-number revealed that bundle-like structures consisting of several aerial hyphae were often formed. S1 nuclease protection analyses were performed to identify the transcriptional start site in the promoters preceding sasA and sasR. The promoter preceding sasA was highly active during vegetative growth. Null mutants for sasABCD among the S. griseus and S. coelicolor A3(2) cells exhibited bald phenotypes; this suggested a positive regulatory role of this gene cluster in the onset of morphogenesis in these two phylogenetically distinct Streptomyces species.


Global and specific responses of the histone acetylome to systematic perturbation.
Regulation of histone acetylation is fundamental to the utilization of eukaryotic genomes in chromatin. Aberrant acetylation contributes to disease and can be clinically combated by inhibiting the responsible enzymes. Our knowledge of the histone acetylation system is patchy because we so far lacked the methodology to describe acetylation patterns and their genesis by integrated enzyme activities. We devised a generally applicable, mass spectrometry-based strategy to precisely and accurately quantify combinatorial modification motifs. This was applied to generate a comprehensive inventory of acetylation motifs on histones H3 and H4 in Drosophila cells. Systematic depletion of known or suspected acetyltransferases and deacetylases revealed specific alterations of histone acetylation signatures, established enzyme-substrate relationships, and unveiled an extensive crosstalk between neighboring modifications. Unexpectedly, overall histone acetylation levels remained remarkably constant upon depletion of individual acetyltransferases. Conceivably, the acetylation level is adjusted to maintain the global charge neutralization of chromatin and the stability of nuclei.


PKD1, PKD2, and their substrate Kidins220 regulate neurotensin secretion in the BON human endocrine cell line.
Neurotensin (NT) is a gut peptide that plays an important role in gastrointestinal secretion, motility, and growth as well as the proliferation of NT receptor-positive cancers. Protein kinase D (PKD) family members (PKD1, 2, and 3) have been identified as important regulators of secretory transport at the trans-Golgi network. Previously, we showed that PKD1 contributes to stimulated NT secretion; however, the mechanisms are not entirely clear. Here, we show that Kidins220, which is a substrate of PKD proteins in neuroendocrine cells, is localized in the ends of the processes of BON cells, similar to the expression pattern of NT vesicles, and translocates to the membrane and large vesicle-like structures formed in response to phorbol 12-myristate 13-acetate treatment. The short hairpin RNA targeting Kidins220 inhibits NT secretion in parental BON cells or BON cells stably expressing the gastrin-releasing peptide receptor treated with either phorbol 12-myristate 13-acetate or bombesin, respectively. Furthermore, we demonstrate that endogenous PKD1, PKD2, and Kidins220 co-exist with NT-containing vesicles. Overexpression of the kinase-dead PKD1 abrogates Kidins220 expression and NT vesicle formation. Our data establish a physiological link between the PKD/Kidins220 pathway and NT-containing vesicles and suggest the role of this pathway in the regulation of hormone secretion. Because NT is an important gut hormone that affects secretion, inflammation, and both normal and tumor cell growth, our findings identify a novel signaling pathway that may be amenable to drug targeting for clinical applications.


Comparison of inducibility of multidrug resistance (MDR)1, multidrug resistance-associated protein (MRP)1, and MRP2 mRNAs by prototypical microsomal enzyme inducers in primary cultures of human and cynomolgus monkey hepatocytes.
This study investigated the changes in the mRNA levels of the ATP binding cassette (ABC) transporters multidrug resistance 1 (MDR1), multidrug resistance-associated protein 1 (MRP1), and multidrug resistance-associated protein 2 (MRP2) following exposure to the prototypical microsomal enzyme inducers rifampicin (Rif), dexamethasone (Dex), and omeprazole (Ome) in primary cultures of cryopreserved human and cynomolgus monkey hepatocytes. Analysis was performed by real-time reverse transcription-polymerase chain reaction using primers and TaqMan probes. First, the time course of the mRNA expression of these transporters in primary cultures of human and cynomolgus monkey hepatocytes was examined in detail. The ratio of MDR1 and MRP2 mRNA to beta-actin mRNA in both human and cynomolgus monkey hepatocytes remained constant from 48 to 72 h and from 24 to 72 h of culture, respectively. Second, the hepatocytes were exposed to the inducers and the changes in the levels of the transporter mRNAs were examined. Rif increased MDR1 and MRP1 mRNA levels in both human and cynomolgus monkey hepatocytes, while Ome slightly increased MDR1 and MRP1 mRNA levels in cynomolgus monkey hepatocytes. Rif and Ome increased MRP2 mRNA levels in both human and cynomolgus monkey hepatocytes. In contrast, Dex tended to decrease the mRNA levels of MDR1, MRP1, and MRP2 in both human and cynomolgus monkey hepatocytes. Cynomolgus monkey hepatocytes appeared to be more responsive than human hepatocytes to the inducers. These results indicate that primary cultures of cynomolgus monkey hepatocytes are as useful as primary cultures of human hepatocytes for evaluating the induction of MDR1, MRP1, and MRP2 mRNAs in preclinical studies.


Molecular analysis of rifampin-resistant Mycobacterium tuberculosis isolated from Korea by polymerase chain reaction-single strand conformation polymorphism sequence analysis.
To assess the molecular mechanism of rifampin (RMP) resistance in clinical strains of Mycobacterium tuberculosis. The molecular nature of a part of the rpoB gene in 77 M. tuberculosis clinical strains isolated in Korea was analyzed using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and PCR-sequence analysis. Among 67 RMP-resistant isolates, 50 showed SSCP profiles different from that of an RMP-sensitive control strain, M. tuberculosis H37Rv, indicating the possible existence of a sequence alteration in this region of the rpoB gene, while 17 resistant isolates displayed SSCP profiles indistinguishable from that of the sensitive control strain. Subsequently, 17 clinical isolates whose SSCP profiles were difficult to distinguish from the control strain were subjected to sequence analysis. The analysis revealed that all 17 isolates did indeed contain mutations in the 81 bp region of the rpoB gene, which is associated with RMP resistance. The results from our study clearly indicate that the molecular mechanism of RMP resistance in M. tuberculosis isolates from Korea involves alterations in the rpoB gene. In addition, this study suggests that PCR-direct sequence analysis works more efficiently and accurately than PCR-SSCP analysis for rapid screening of RMP-resistant M. tuberculosis clinical isolates.


The European Portuguese version of the Oxford Cognitive Screening (OCS-Pt): a screening test for acute stroke patients.
In Portugal, there is no gold standard test to assess acute stroke patients in the hospital context. However, cognitive deficits are common after a stroke episode and are followed by a negative impact on a patient's quality of life and rehabilitation. Here, we first aimed to adapt the Oxford Cognitive Screening to European Portuguese speakers (OCS-Pt), develop normative data cut-offs, and report the psychometric properties of the OCS-Pt. The second aim was to test the incidence of impairments in acute stroke patients. We tested 137 healthy participants aged between 25 and 92 years old, and we report normative cut-offs based on the 5th and 95th percentile, and on the patterns for age and education level. Our results are similar to other European OCS versions, and results using convergent and divergent validity show satisfactory values ranged between moderate to high. Additionally, 146 acute stroke patients, in the first week after the stroke episode, performed the OCS-Pt. Results show that this is an inclusive test and allows to discriminate between impaired and preserved functions. In conclusion, OCS-Pt is a promising cognitive screening tool to assess acute stroke survivors to be used in stroke care in Portugal.


Somatic VHL gene alterations in MEN2-associated medullary thyroid carcinoma.
Germline mutations in RET are responsible for multiple endocrine neoplasia type 2 (MEN2), an autosomal dominantly inherited cancer syndrome that is characterized by medullary thyroid carcinoma (MTC), pheochromocytoma, and parathyroid hyperplasia/adenoma. Recent studies suggest a "second hit" mechanism resulting in amplification of mutant RET. Somatic VHL gene alterations are implicated in the pathogenesis of MEN2 pheochromocytomas. We hypothesized that somatic VHL gene alterations are also important in the pathogenesis of MEN2-associated MTC. We analyzed 6 MTCs and 1 C-cell hyperplasia (CCH) specimen from 7 patients with MEN2A and RET germline mutations in codons 609, 618, 620, or 634, using microdissection, microsatellite analysis, phosphorimage densitometry, and VHL mutation analysis. First, we searched for allelic imbalance between mutant and wild-type RET by using the polymorphic markers D10S677, D10S1239, and RET on thyroid tissue from these patients. Evidence for RET amplification by this technique could be demonstrated in 3 of 6 MTCs. We then performed LOH analysis using D3S1038 and D3S1110 which map to the VHL gene locus at 3p25/26. VHL gene deletion was present in 3 MTCs. These 3 MTCs also had an allelic imbalance between mutant and wild-type RET. Mutation analysis of the VHL gene showed a somatic frameshift mutation in 1 MTC that also demonstrated LOH at 3p25/26. In the 2 other MTCs with allelic imbalance of RET and somatic VHL gene deletion, no somatic VHL mutation could be detected. The CCH specimen did neither reveal RET imbalance nor somatic VHL gene alterations. These data suggest that a RET germline mutation is necessary for development of CCH, that allelic imbalance between mutant and wild-type RET may set off tumorigenesis, and that somatic VHL gene alterations may not play a major role in tumorigenesis of MEN2A-associated MTC.


Effect of graft choice on the outcome of revision anterior cruciate ligament reconstruction in the Multicenter ACL Revision Study (MARS) Cohort.
Most surgeons believe that graft choice for anterior cruciate ligament (ACL) reconstruction is an important factor related to outcome; however, graft choice for revision may be limited due to previously used grafts. Autograft use would result in increased sports function, increased activity level, and decreased osteoarthritis symptoms (as measured by validated patient-reported outcome instruments). Autograft use would result in decreased graft failure and reoperation rate 2 years after revision ACL reconstruction. Cohort study; Level of evidence, 2. Patients undergoing revision ACL reconstruction were identified and prospectively enrolled by 83 surgeons at 52 sites. Data collected included baseline demographics, surgical technique, pathologic abnormalities, and the results of a series of validated, patient-reported outcome instruments (International Knee Documentation Committee [IKDC], Knee injury and Osteoarthritis Outcome Score [KOOS], Western Ontario and McMaster Universities Osteoarthritis Index [WOMAC], and Marx activity rating score). Patients were followed up at 2 years and asked to complete the identical set of outcome instruments. Incidences of additional surgery and reoperation due to graft failure were also recorded. Multivariate regression models were used to determine the predictors (risk factors) of IKDC, KOOS, WOMAC, Marx scores, graft rerupture, and reoperation rate at 2 years after revision surgery. A total of 1205 patients (697 [58%] males) were enrolled. The median age was 26 years. In 88% of patients, this was their first revision, and 341 patients (28%) were undergoing revision by the surgeon who had performed the previous reconstruction. The median time since last ACL reconstruction was 3.4 years. Revision using an autograft was performed in 583 patients (48%), allograft was used in 590 (49%), and both types were used in 32 (3%). Questionnaire follow-up was obtained for 989 subjects (82%), while telephone follow-up was obtained for 1112 (92%). The IKDC, KOOS, and WOMAC scores (with the exception of the WOMAC stiffness subscale) all significantly improved at 2-year follow-up (P < .001). In contrast, the 2-year Marx activity score demonstrated a significant decrease from the initial score at enrollment (P < .001). Graft choice proved to be a significant predictor of 2-year IKDC scores (P = .017). Specifically, the use of an autograft for revision reconstruction predicted improved score on the IKDC (P = .045; odds ratio [OR] = 1.31; 95% CI, 1.01-1.70). The use of an autograft predicted an improved score on the KOOS sports and recreation subscale (P = .037; OR = 1.33; 95% CI, 1.02-1.73). Use of an autograft also predicted improved scores on the KOOS quality of life subscale (P = .031; OR = 1.33; 95% CI, 1.03-1.73). For the KOOS symptoms and KOOS activities of daily living subscales, graft choice did not predict outcome score. Graft choice was a significant predictor of 2-year Marx activity level scores (P = .012). Graft rerupture was reported in 37 of 1112 patients (3.3%) by their 2-year follow-up: 24 allografts, 12 autografts, and 1 allograft and autograft. Use of an autograft for revision resulted in patients being 2.78 times less likely to sustain a subsequent graft rupture compared with allograft (P = .047; 95% CI, 1.01-7.69). Improved sports function and patient-reported outcome measures are obtained when an autograft is used. Additionally, use of an autograft shows a decreased risk in graft rerupture at 2-year follow-up. No differences were noted in rerupture or patient-reported outcomes between soft tissue and bone-patellar tendon-bone grafts. Surgeon education regarding the findings of this study has the potential to improve the results of revision ACL reconstruction.


IMproving the practice of intrapartum electronic fetal heart rate MOnitoring with cardiotocography for safer childbirth (the IMMO programme): protocol for a qualitative study.
Suboptimal electronic fetal heart rate monitoring (EFM) in labour using cardiotocography (CTG) has been identified as one of the most common causes of avoidable harm in maternity care. Training staff is a frequently proposed solution to reduce harm. However, current approaches to training are heterogeneous in content and format, making it difficult to assess effectiveness. Technological solutions, such as digital decision support, have not yet demonstrated improved outcomes. Effective improvement strategies require in-depth understanding of the technical and social mechanisms underpinning the EFM process. The aim of this study is to advance current knowledge of the types of errors, hazards and failure modes in the process of classifying, interpreting and responding to CTG traces. This study is part of a broader research programme aimed at developing and testing an intervention to improve intrapartum EFM. The study is organised into two workstreams. First, we will conduct observations and interviews in three UK maternity units to gain an in-depth understanding of how intrapartum EFM is performed in routine clinical practice. Data analysis will combine the insights of an ethnographic approach (focused on the social norms and interactions, values and meanings that appear to be linked with the process of EFM) with a systems thinking approach (focused on modelling processes, actors and their interactions). Second, we will use risk analysis techniques to develop a framework of the errors, hazards and failure modes that affect intrapartum EFM. This study has been approved by the West Midlands-South Birmingham Research Ethics Committee, reference number: 18/WM/0292. Dissemination will take the form of academic articles in peer-reviewed journals and conferences, along with tailored communication with various stakeholders in maternity care.


Quality of life and burden in caregivers of youth with obsessive-compulsive disorder presenting for intensive treatment.
Pediatric obsessive-compulsive disorder (OCD) is associated with deleterious familial effects; caregivers are often enmeshed in the disorder and can experience considerable burden and decreased quality of life (QoL). Consequently, this study examined burden and QoL in caregivers of youth with OCD enrolled in an intensive outpatient or partial hospitalization program. The relationships between caregiver QoL and burden and the following variables were investigated: OCD symptom severity, functioning (youth functional impairment, general family functioning), family (family accommodation, parental relationship satisfaction, positive aspects of caregiving), and comorbid psychopathology (caregiver anxiety and depressive symptoms, youth internalizing and externalizing behaviors). Seventy-two child and caregiver dyads completed clinician- and self-rated questionnaires. Components of caregiver QoL correlated with caregiver-rated functional impairment, family accommodation, youth externalizing behaviors, and caregiver psychopathology. Aspects of caregiver burden correlated with child OCD symptom severity, functional impairment related to OCD, as well as caregiver and child comorbid psychopathology. Caregiver depressive symptoms predicted caregiver QoL, and caregiver depressive symptoms and child externalizing symptoms both predicted caregiver burden. Caregiver burden did not mediate the relationship between obsessive-compulsive symptom severity and caregiver QoL. Ultimately, elucidating factors associated with increased caregiver burden and poorer QoL is pertinent for identifying at-risk families and developing targeted interventions.


Refolding kinetics of denatured-reduced lysozyme in the presence of folding aids.
The refolding kinetic behavior of denatured-reduced lysozyme in the presence of folding aids (acetamide, acetone, thiourea, L-arginine or glycerol) was studied utilizing a simplified model describing the competition between first-order folding reaction and third-order aggregation. It was found that the protein folding aids could be categorized into two groups. One of them at proper concentrations, such as acetamide, acetone, thiourea and L-arginine, stabilized unfolded protein or folding intermediates. In the presence of these additives, the folding rate decreased with increasing their concentration, and there existed a concentration where the aggregation rate constant was minimized. So, there was an optimum concentration for the folding aids to produce a high yield. The other group was protein stabilizers such as glycerol. In the presence of this kind of folding aids, both the refolding rate and yield were enhanced by increasing their concentration to a proper value. Moreover, their effect on improving protein refolding was additive to those of the first group. So the cooperative application of the two kinds of folding aids could result in favorable refolding rate and yield of protein.


Risk of melanoma and vitamin A, coffee and alcohol: a case-control study from Italy.
Limited information is available on the association between the risk of cutaneous malignant melanoma (CMM) and dietary factors. The issue was investigated using data from a case-control study conducted in Italy between 1992 and 1994, including 542 patients with incident, histologically confirmed CMM and 538 controls, admitted to the same hospitals as cases for non-dermatologic and non-neoplastic diseases. We found a significant inverse association between vitamin A intake and CMM risk. The multivariate odds ratio, after allowance for phenotypic characteristics and history of sunburns, for the highest, compared with the lowest, quartile of intake was 0.71 (95% confidence interval (CI) 0.50-1.02) for beta-carotene, 0.57 (95% CI 0.39-0.83) for retinol, and 0.51 (95% CI 0.35-0.74) for total vitamin A. We found no appreciable association of CMM risk with selected food items, including fish, meat, vegetables, fruit, dairy products, wholemeal bread, alcohol, coffee and tea drinking. Consumption of tea appeared to have a protective effect on CMM risk. The relations with measures of dietary vitamin A are, however, moderate compared with the associations between phenotypic characteristics, sun exposure and number of naevi and CMM risk.


Optimal Hemodynamics During Left Ventricular Assist Device Support Are Associated With Reduced Readmission Rates.
Left ventricular assist device (LVAD) therapy improves the hemodynamics of advanced heart failure patients. However, it is unknown whether hemodynamic optimization improves clinical outcomes. The aim of this study was to investigate whether hemodynamic optimization reduces hospital readmission rate in LVAD patients. LVAD patients undergoing an invasive hemodynamic ramp test were prospectively enrolled and followed for 1 year. LVAD speed was optimized using a ramp test, targeting the following goals: central venous pressure <12 mm Hg, pulmonary capillary wedge pressure <18 mm Hg, and cardiac index >2.2 L/(min·m2). The frequency and cause of hospital readmissions were compared between patients who achieved (optimized group) or did not achieve (nonoptimized group) these goals. Eighty-eight outpatients (median 61 years old, 53 male) underwent ramp testing 236 days after LVAD implantation, and 54 (61%) had optimized hemodynamics after LVAD speed adjustment. One-year survival after the ramp study was comparable in both groups (89% versus 88%). The total hospital readmission rate was lower in the optimized group compared with the nonoptimized group (1.15 versus 2.86 events/y, P<0.001). This result was predominantly because of a reduction in the heart failure readmission rate in the optimized group (0.08 versus 0.71 events/y, P=0.016). LVAD patients, in whom hemodynamics were optimized, had a significantly lower rate of hospital readmissions, primarily because of fewer heart failure admissions. These findings highlight the importance of achieving hemodynamic optimization in LVAD patients.


Using Aerial and Vehicular NFV Infrastructures to Agilely Create Vertical Services.
5G communications have become an enabler for the creation of new and more complex networking scenarios, bringing together different vertical ecosystems. Such behavior has been fostered by the network function virtualization (NFV) concept, where the orchestration and virtualization capabilities allow the possibility of dynamically supplying network resources according to its needs. Nevertheless, the integration and performance of heterogeneous network environments, each one supported by a different provider, and with specific characteristics and requirements, in a single NFV framework is not straightforward. In this work we propose an NFV-based framework capable of supporting the flexible, cost-effective deployment of vertical services, through the integration of two distinguished mobile environments and their networks: small sized unmanned aerial vehicles (SUAVs), supporting a flying ad hoc network (FANET) and vehicles, promoting a vehicular ad hoc network (VANET). In this context, a use case involving the public safety vertical will be used as an illustrative example to showcase the potential of this framework. This work also includes the technical implementation details of the framework proposed, allowing to analyse and discuss the delays on the network services deployment process. The results show that the deployment times can be significantly reduced through a distributed VNF configuration function based on the publish-subscribe model.


Coffee Consumption and the Risk of Depression in a Middle-Aged Cohort: The SUN Project.
Coffee is one of the most widely consumed drinks around the world, while depression is considered the major contributor to the overall global burden of disease. However, the investigation on coffee consumption and depression is limited and results may be confounded by the overall dietary pattern. We assessed the relationship between coffee intake and the risk of depression, controlling for adherence to the Mediterranean diet. We studied 14,413 university graduates of the 'Seguimiento Universidad de Navarra' (SUN) cohort, initially free of depression. We evaluated coffee consumption using a validated food-frequency questionnaire (FFQ). Incident depression cases were adjudicated only if the participant met two criteria simultaneously: (a) validated physician-diagnosed depression together with (b) new onset of habitual antidepressant use. Both criteria were needed; participants meeting only one of them were not classified as cases. Participants who drank at least four cups of coffee per day showed a significantly lower risk of depression than participants who drank less than one cup of coffee per day (HR: 0.37 (95% CI 0.15⁻0.95)). However, overall, we did not observe an inverse linear dose⁻response association between coffee consumption and the incidence of depression (p for trend = 0.22).


Early events in innate immunity in the recognition of microbial pathogens.
Innate immunity is characterised by a rapid action of host effector molecules and leukocytes aimed at limiting the multiplication of invading microbial organisms and destroying them. The recognition and destruction of microorganisms involves humoral factors (e.g., the complement system and natural antibodies) and different cell types (e.g., phagocytic cells, mast cells, natural killer cells). Microbial detection by cells involves germ line-encoded pattern-recognition receptors such as Toll-like receptors and nucleotide-binding oligomerization domain-like receptors. Cellular activation by pathogens leads to the release of antimicrobial peptides (e.g., defensins and peptidoglycan recognition proteins) and cytokines that orchestrate the anti-infectious response. Cytokines enhance phagocytosis and leukocyte microbicidal activity, allow cellular recruitment into the infectious focus, boost hematopoiesis, induce fever and lead to the production of acute phase proteins.


Analysis of signaling events by dynamic phosphoflow cytometry.
Many proteins involved in cell signaling are phosphorylated. To determine the phosphorylation status of these signaling molecules at the single-cell level, we present a protocol for using state-specific antibodies to detect target phosphoproteins with fluorescence measurements by flow cytometry. To improve the signal intensity, a sandwich-labeling method for the analysis of signaling proteins is performed. By comparing the phosphorylation state of proteins in the presence and absence of sodium pervanadate, a nonspecific tyrosine phosphatase inhibitor, we determined the relative amount of tyrosine-phosphorylated protein in the samples, which reflects the activity of the signaling pathway. This dynamic approach, in combination with the signal amplification through a sandwich-labeling method, produces accurate and reproducible measurement of the activity of signaling pathways.


Information and support needs during recovery from postpartum psychosis.
Postpartum Psychosis (PP) is a severe and debilitating psychiatric illness with acute onset in the days following childbirth. Recovering from an episode can be a long and difficult process. The aim of this study was to gain an understanding of the difficulties faced by recovering women and to inform the planning of post-discharge information and support services. A study was designed collaboratively by service user and academic researchers. Women with experience of PP were trained in qualitative research methodology. Service user researchers (SURs) led in-depth interviews into women's experiences of recovery. PP is a life-changing experience that challenges women's sense of personal and social identity. Recovery themes are organised around ruminating and rationalising, rebuilding social confidence, gaining appropriate health service support, the facilitation of family functioning, obtaining appropriate information, and understanding that recovery will take time. Women suffering from PP must be adequately supported following discharge from psychiatric hospital if we are to address maternal suicide rates. We describe a successful collaboration between academics and service users exploring the needs of women and their families.


Membrane protein structure determination - the next generation.
The field of Membrane Protein Structural Biology has grown significantly since its first landmark in 1985 with the first three-dimensional atomic resolution structure of a membrane protein. Nearly twenty-six years later, the crystal structure of the beta2 adrenergic receptor in complex with G protein has contributed to another landmark in the field leading to the 2012 Nobel Prize in Chemistry. At present, more than 350 unique membrane protein structures solved by X-ray crystallography (http://blanco.biomol.uci.edu/mpstruc/exp/list, Stephen White Lab at UC Irvine) are available in the Protein Data Bank. The advent of genomics and proteomics initiatives combined with high-throughput technologies, such as automation, miniaturization, integration and third-generation synchrotrons, has enhanced membrane protein structure determination rate. X-ray crystallography is still the only method capable of providing detailed information on how ligands, cofactors, and ions interact with proteins, and is therefore a powerful tool in biochemistry and drug discovery. Yet the growth of membrane protein crystals suitable for X-ray diffraction studies amazingly remains a fine art and a major bottleneck in the field. It is often necessary to apply as many innovative approaches as possible. In this review we draw attention to the latest methods and strategies for the production of suitable crystals for membrane protein structure determination. In addition we also highlight the impact that third-generation synchrotron radiation has made in the field, summarizing the latest strategies used at synchrotron beamlines for screening and data collection from such demanding crystals. This article is part of a Special Issue entitled: Structural and biophysical characterisation of membrane protein-ligand binding.


The quest for One Health: human resource training aspects.
Appropriately trained Human Resources for Health (HRH) are key inputs into One Health. '… more than 50% of all infectious diseases of humans originate from animals and that, of the emerging diseases about 75% could be traced back to animal origin' (Rweyemamu et al. 2006). A comprehensive understanding of the social determinants of health, through an appropriate training model for HRH, is a key input. This study aimed to explore if human and veterinary medical schools were using such a model or providing time for this model in their curricula. Specific objectives were to: determine the time that human and veterinary medical schools' curricula provide for subjects or courses related to the social determinants of health; analyse the curricula contents to establish how they relate to the social determinants of health; and explore how a bio-medical model may influence the graduates' understanding and practice of One Health. A review of human and veterinary graduate-level medical schools' curricula in East Africa was performed in April 2013 and May 2013. The findings were: in the curricula, SDH contents for knowledge enhancement about One Health are minimal and that teaching is Germ Theory model-driven and partisan. Out of the total training time for physicians and veterinarians, less than 10% was provided for the social determinants of health-related courses. In conclusion, the curricula and training times provided are inadequate for graduates to fully understand the social determinants of health and their role in One Health. Furthermore, the Germ Theory model that has been adopted addresses secondary causes and is inappropriate. There is a need for more in-depth model. This article suggests that a vicious cycle of ill-health model must be taught.


Information and Risk Modification Trial (INFORM): design of a randomised controlled trial of communicating different types of information about coronary heart disease risk, alongside lifestyle advice, to achieve change in health-related behaviour.
Cardiovascular disease (CVD) remains the leading cause of death globally. Primary prevention of CVD requires cost-effective strategies to identify individuals at high risk in order to help target preventive interventions. An integral part of this approach is the use of CVD risk scores. Limitations in previous studies have prevented reliable inference about the potential advantages and the potential harms of using CVD risk scores as part of preventive strategies. We aim to evaluate short-term effects of providing different types of information about coronary heart disease (CHD) risk, alongside lifestyle advice, on health-related behaviours. In a parallel-group, open randomised trial, we are allocating 932 male and female blood donors with no previous history of CVD aged 40-84 years in England to either no intervention (control group) or to one of three active intervention groups: i) lifestyle advice only; ii) lifestyle advice plus information on estimated 10-year CHD risk based on phenotypic characteristics; and iii) lifestyle advice plus information on estimated 10-year CHD risk based on phenotypic and genetic characteristics. The primary outcome is change in objectively measured physical activity. Secondary outcomes include: objectively measured dietary behaviours; cardiovascular risk factors; current medication and healthcare usage; perceived risk; cognitive evaluation of provision of CHD risk scores; and psychological outcomes. The follow-up assessment takes place 12 weeks after randomisation. The experiences, attitudes and concerns of a subset of participants will be also studied using individual interviews and focus groups. The INFORM study has been designed to provide robust findings about the short-term effects of providing different types of information on estimated 10-year CHD risk and lifestyle advice on health-related behaviours. Current Controlled Trials ISRCTN17721237 . Registered 12 January 2015.


Long-term recovery patterns and limited spillover of large predatory fish in a Mediterranean MPA.
Based on 19 y of visual census data from the Medes Islands MPA (NW Mediterranean), this study analyzes the carrying capacity (K) and population recovery time of six species of fish strongly affected by harvesting pressure along the Mediterranean coast. Three of these species (Epinephelus marginatus, Diplodus cervinus and Dicentrachus labrax) have practically reached carrying capacity in the Medes Islands MPA, while others are still approaching population stabilization (Sciaena umbra) or are still increasing in biomass (Dentex dentex). The one exception to these trends is S. aurata, which tended to decrease inside the MPA, probably due to fishing just outside its borders. These results confirm that fish populations may require decadal time scales to recover from exploitation, both in terms of total abundance (21 to 29 y to exceed 95% K) as well as total biomass (25 to 35 y), and that rates of recovery differ between species (13 to 31 y). The recovery and saturation observed within the no-take zone contrasts with results obtained in the partially protected buffer area and the peripheral area open for fishing, which show much lower biomass values. In general, the spillover from the MPA is very moderate, and its effects extend only to the partially protected area.


Carcinogenic activity of a chlorinated polyether polyurethan.
On the basis of the results of an earlier study, a particular polyurethan sample (Y-238) was selected for further evaluation of its carcinogenic potential. This sample was subjected to physical and chemical tests for elucidation of its chemical structure, molecular weight, and molecular weight distribution. Additional biological tests were conducted on male NBR rats by implanting various quantities of the sample i.p., while others received an intrabronchus implant. Tumors, assessed histologically as malignant, were observed following both routes of implantation. The most common neoplasms of the pulmonary site was epidermoid carcinoma, while fibrosarcoma was the most common neoplasm in the peritoneal cavity. Data from the i.p. implantation suggested a dose-related incidence of cancers.


Synthesis and biological evaluation of prodigiosene conjugates of porphyrin, estrone and 4-hydroxytamoxifen.
To generate the first series of prodigiosene conjugates, the tripyrrolic skeleton was appended to estrone, tamoxifen and porphyrin frameworks by way of ester linkers and various hydrocarbon chain lengths. The ability of the conjugates to inhibit various types of cancer cells was evaluated in vitro. The porphyrin conjugates did not exhibit significant activity. The estrone conjugates exhibited modest activity, for the most part. However, significantly greater growth inhibition activity against certain breast, colon, lung, leukemia, melanoma and prostate cell lines was noted. This unusual effect for this first generation model class of compound warrants further investigation and comparison to cases where estrogens are linked to prodigiosenes via connection points that do not feature in estrogen receptor binding. The 4-hydroxytamoxifen conjugates exhibit nanomolar range activity against the MCF-7 breast cancer cell line, paving the way to expand the scope and connectivity of prodigiosene-tamoxifen conjugates.


Regional differences in glutaminase activation by phosphate and calcium in rat brain: impairment in aged rats and implications for regional glutaminase isozymes.
Regional regulation of glutaminase by phosphate and calcium was examined in the temporal cortex (TCX), striatum (STR) and hippocampus (HIPP) from adult and aged male F344 rats. Phosphate-dependent glutaminase activity in adult rats was significantly lower (35-43%) in the HIPP (100 and 150 mM) and STR (150 mM) compared to PAG activity in the TCX. Phosphate activation in aged rats was 50-60% lower in the HIPP at concentrations greater than 25 mM compared to the aged TCX or STR. PAG activity in the TCX and STR was unaffected by age, but was significantly reduced (30-50%) in the HIPP from aged rats at phosphate concentrations of 25 mM and greater when compared to adult rats. In adult rats at concentrations of CaCl2 above 1 mM, PAG activity was significantly lower (60-75%) in the STR and HIPP when compared to the TCX. In aged rats, PAG activity (1 mM CaCl2) in the HIPP was significantly less (50%) than STR PAG activity in aged rats. Diminished PAG activity was seen only in the TCX (2.5 mM; 32%), and the HIPP (0.5 mM; 25% and 1 mM; 38%) at higher calcium concentrations compared to adult. Phosphate-independent calcium activation of PAG occurred in the HIPP but not in either the TCX or the STR. Addition of phosphate resulted in a synergistic activation of PAG in the STR and TCX, but not in the HIPP. These findings suggest that PAG is regionally regulated by phosphate and calcium, and this regulation is impaired in aged rats. These data also support the hypothesis that isozymes of PAG exist with different regulatory properties.