Assessing the role of undetected colonization and isolation precautions in reducing methicillin-resistant Staphylococcus aureus transmission in intensive care units.
Screening and isolation are central components of hospital methicillin-resistant Staphylococcus aureus (MRSA) control policies. Their prevention of patient-to-patient spread depends on minimizing undetected and unisolated MRSA-positive patient days. Estimating these MRSA-positive patient days and the reduction in transmission due to isolation presents a major methodological challenge, but is essential for assessing both the value of existing control policies and the potential benefit of new rapid MRSA detection technologies. Recent methodological developments have made it possible to estimate these quantities using routine surveillance data. Colonization data from admission and weekly nares cultures were collected from eight single-bed adult intensive care units (ICUs) over 17 months. Detected MRSA-positive patients were isolated using single rooms and barrier precautions. Data were analyzed using stochastic transmission models and model fitting was performed within a Bayesian framework using a Markov chain Monte Carlo algorithm, imputing unobserved MRSA carriage events. Models estimated the mean percent of colonized-patient-days attributed to undetected carriers as 14.1% (95% CI (11.7, 16.5)) averaged across ICUs. The percent of colonized-patient-days attributed to patients awaiting results averaged 7.8% (6.2, 9.2). Overall, the ratio of estimated transmission rates from unisolated MRSA-positive patients and those under barrier precautions was 1.34 (0.45, 3.97), but varied widely across ICUs. Screening consistently detected >80% of colonized-patient-days. Estimates of the effectiveness of barrier precautions showed considerable uncertainty, but in all units except burns/general surgery and one cardiac surgery ICU, the best estimates were consistent with reductions in transmission associated with barrier precautions.


Prevalence and Factors Associated with Mental and Emotional Health Outcomes among Africans during the COVID-19 Lockdown Period-A Web-based Cross-Sectional Study.
Mental health and emotional responses to the effects of COVID-19 lockdown in sub-Saharan Africa (SSA) are of serious public health concern and may negatively affect the mental health status of people. Hence, this study assessed the prevalence of mental health symptoms as well as emotional reactions among sub-Saharan Africans (SSAs) and associated factors among SSAs during the COVID-19 lockdown period. This was a web-based cross-sectional study on mental health and emotional features from 2005 respondents in seven SSA countries. This study was conducted between 17 April and 17 May 2020 corresponding to the lockdown period in most SSA countries. Respondents aged 18 years and above and the self-reported symptoms were feeling anxious, being worried, angry, bored and frustrated. These were the main outcomes and were treated as dichotomous variables. Univariate and multivariate logistic regression analyses were used to identify the factors associated with these symptoms. We found that over half (52.2%) of the participants reported any of the mental health symptoms and the prevalence of feeling bored was 70.5% followed by feeling anxious (59.1%), being worried (57.5%), frustrated (51.5%) and angry (22.3%) during the COVID-19 pandemic. Multivariate analysis revealed that males, those aged >28 years, those who lived in Central and Southern Africa, those who were not married, the unemployed, those living with more than six persons in a household, had higher odds of mental health and emotional symptoms. Similarly, people who perceived low risk of contracting the infection, and those who thought the pandemic would not continue after the lockdown had higher odds of mental health and emotional symptoms. Health care workers had lower odds for feeling angry than non-healthcare workers. During the COVID-19 lockdown periods in SSA, about one in two participants reported mental health and emotional symptoms. Public health measures can be effectively used to identify target groups for prevention and treatment of mental health and emotional symptoms. Such interventions should be an integral component of SSA governments' response and recovery strategies of any future pandemic.


SCOP2 prototype: a new approach to protein structure mining.
We present a prototype of a new structural classification of proteins, SCOP2 (http://scop2.mrc-lmb.cam.ac.uk/), that we have developed recently. SCOP2 is a successor to the Structural Classification of Proteins (SCOP, http://scop.mrc-lmb.cam.ac.uk/scop/) database. Similarly to SCOP, the main focus of SCOP2 is to organize structurally characterized proteins according to their structural and evolutionary relationships. SCOP2 was designed to provide a more advanced framework for protein structure annotation and classification. It defines a new approach to the classification of proteins that is essentially different from SCOP, but retains its best features. The SCOP2 classification is described in terms of a directed acyclic graph in which nodes form a complex network of many-to-many relationships and are represented by a region of protein structure and sequence. The new classification project is expected to ensure new advances in the field and open new areas of research.


What do implicit measures measure?
We identify several ongoing debates related to implicit measures, surveying prominent views and considerations in each. First, we summarize the debate regarding whether performance on implicit measures is explained by conscious or unconscious representations. Second, we discuss the cognitive structure of the operative constructs: are they associatively or propositionally structured? Third, we review debates about whether performance on implicit measures reflects traits or states. Fourth, we discuss the question of whether a person's performance on an implicit measure reflects characteristics of the person who is taking the test or characteristics of the situation in which the person is taking the test. Finally, we survey the debate about the relationship between implicit measures and (other kinds of) behavior. This article is categorized under: Philosophy > Representation Psychology > Theory and Methods Psychology > Prediction Philosophy > Psychological Capacities.


Distinct Modulations in Sensorimotor Postmovement and Foreperiod β-Band Activities Related to Error Salience Processing and Sensorimotor Adaptation.
In a recent study, Tan et al. (2014a,b) showed that the increase in β-power typically observed after a movement above sensorimotor regions (β-rebound) is attenuated when movement-execution errors are induced by visual perturbations. Moreover, akin to sensorimotor adaptation, the effect depended on the context in which the errors are experienced. Thus the β-rebound attenuation might relate to neural processes involved in trial-to-trial adaptive mechanisms. In two EEG experiments with human participants, along with the β-rebound, we examine β-activity during the preparation of reaches immediately following perturbed movements. In the first experiment, we show that both foreperiod and postmovement β-activities are parametrically modulated by the sizes of kinematic errors produced by unpredictable mechanical perturbations (force field) independent of their on-line corrections. In the second experiment, we contrast two types of reach errors: movement-execution errors that trigger trial-to-trial adaptive mechanisms and goal errors that do not elicit sensorimotor adaptation. Movement-execution errors were induced by mechanical or visual perturbations, whereas goal errors were caused by unexpected displacements of the target at movement initiation. Interestingly, foreperiod and postmovement β-activities exhibit contrasting patterns, pointing to important functional differences of their underlying neuronal activity. While both types of reach errors attenuate the postmovement β-rebound, only the kinematic errors that trigger trial-to-trial motor-command updates influenced β-activity during the foreperiod. These findings suggest that the error-related modulation of the β-rebound may reflect salience processing, independent of sensorimotor adaptation. In contrast, modulations in the foreperiod β-power might relate to the motor-command adjustments activated after movement-execution errors are experienced. The functional significance of sensorimotor β-band (15-25 Hz) oscillations remains uncertain. Recently β-power was found to be reduced following erroneous movements. We extend and refine this novel finding in two crucial ways. First, by contrasting the EEG correlates of movement errors driving or not driving adaptation we dissociate error-salience processing from error-based adaptation. Second, in addition to β-activity in error trials, we examine β-power during the preparation of the subsequent movements. We find clearly distinct patterns of error-related modulations for β-activities preceding and succeeding movements, highlighting critical functional differences. Postmovement β-power may reflect error-salience processing independent of sensorimotor adaptation. In contrast, modulations in the foreperiod β-band power may directly relate to the motor-command adjustments activated after movement-execution errors are experienced.


Fluorescent reagents for in vitro studies of lipid-linked steps of bacterial peptidoglycan biosynthesis: derivatives of UDPMurNAc-pentapeptide containing d-cysteine at position 4 or 5.
UDPMurNAc-L-Ala-gamma-D-Glu-X-D-Ala-DAla (X = L-Lys or m-DAP) is the cytoplasmic precursor for the lipid-linked cycle of bacterial peptidoglycan biosynthesis, consisting of at least four enzymatic reactions, which are targets for antibacterial agents. Fluorescent derivatives of the UDPMurNAc-pentapeptide labelled at the 3rd, 4th, and 5th position of the peptide chain were prepared chemoenzymatically, in order to study the reactions catalysed by enzymes in this cycle. Derivatives labelled on the epsilon-amino group of the 3rd amino acid (N-dansyl, N-fluorescamine and N-phthalaldehyde) were prepared by chemical modification. Two methods were developed for preparation of analogues of UDPMurNAc-pentapeptide containing D-cysteine at position 4 or 5: either by MurF-catalysed ligation of the UDPMurNAc-tripeptide to synthetic D-Ala-D-Cys or D-Cys-D-Ala dipeptides; or by enzymatic synthesis of D-Ala-D-Cys by ligase VanD. D-Cys-containing UDPMurNAc-pentapeptides were labelled with pyrene maleimide, to give 4-pyrene and 5-pyrene labelled derivatives. The fluorescent UDPMurNAc-pentapeptides were processed as substrates by Escherichia coli MraY or E. coli membranes, giving 1.5-150-fold changes in fluorescence upon transformation to lipid intermediate I. Subsequent processing to lipid intermediate II gave rise only to small changes in fluorescence. Pyrene-labelled lipid intermediates I and II can be generated using Micrococcus flavus membranes, enabling the study of the later lipid-linked steps.


Hearing impairments caused by genetic and environmental factors.
Impairments of hearing and balance are major problems in the field of occupational and environmental health. Such impairments have previously been reported to be caused by genetic and environmental factors. However, their mechanisms have not been fully clarified. On the other hand, the inner ear contains spiral ganglion neurons (SGNs) in the organ of Corti, which serve as the primary carriers of auditory information from sensory cells to the auditory cortex in the cerebrum. Inner ears also contain a vestibule in the vicinity of the organ of Corti-one of the organs responsible for balance. Thus, inner ears could be a good target to clarify the pathogeneses of sensorineural hearing losses and impaired balance. In our previous studies with c-Ret knock-in mice and Endothelin receptor B (Ednrb) knock-out mice, it was found that syndromic hearing losses involved postnatal neurodegeneration of SGNs caused by impairments of c-Ret and Ednrb, which play important roles in neuronal development and maintenance of the enteric nervous system. The organ of Corti and the vestibule in inner ears also suffer from degeneration caused by environmental stresses including noise and heavy metals, resulting in impairments of hearing and balance. In this review, we introduce impairments of hearing and balance caused by genetic and environmental factors and focus on impairments of SGNs and the vestibule in inner ears as the pathogeneses caused by these factors.


A study of intraabdominal pressure modification in "component separation" technique for repair of incisional hernia.
Incisional hernias can be treated with laparoscopic and laparotomic surgery. Laparoscopic surgery can be made without the use of mesh when performing component separation technique. This technique allows to limit the adverse effects to foreign body and promotes a tension-free closure. We studied intravescical pressure changes during intervention in order to quantify intrabdominal pressure. A prospective, non-controlled study (cohort study), was made on thirty patients, treated to repair incisional hernia. Standard panniculectomies and component separation technique were performed in all patients. A standard Foley catheter was inserted in to the bladder in order to measure pressure modification in the peri-/postoperative phases. Statistical significance of modifications of pressure values was evaluated with the Wilcoxon's sum rank test. Bladder pressure increased after hernia repair and skin closure and decreased in the first day after surgery, but without returning to the original values, and these modifications were statistically significant. We study intravescical pressure changes as an indirect measurement of intrabdominal pressure. Intrabdominal pressure cut-off for the rise of complications is 20 mmHg. This technique allows to maintain pressure under dangerous limits and to limits complications.


In vitro assessment of cytotoxicity of resin-based dental restorative materials on WEHI 13 var fibroblasts.
The aim of this study was to assess the cytotoxicity of 4 resin composites' eluates on WEHI 13 Var fibroblasts as they aged in a biologic medium. Cytotoxicity was determined by counting the number of viable cells by trypan blue exclusion. Morphologic changes attributable to cytotoxicity were observed by May-Grunwald-Giemsa cytologic staining and microscopic examination. DNA gel electrophoresis was performed to detect possible genotoxicity and DNA damage. All resin composite eluates, except that for Targis, caused a pronounced cytotoxicity during the first 72 hours that gradually decreased after 2 weeks of aging. Severe morphologic alterations and pronounced DNA damage were also observed. These findings indicate that resin-based dental restorative materials release agents cytotoxic and genotoxic to fibroblasts. Cytotoxicity is gradually decreased as the composite resins age in a biologic-relevant medium.


The relation of secondary sex ratio and miscarriage history with Toxoplasma gondii infection.
Toxoplasma gondii is a protozoan parasite with worldwide distribution, infecting a broad-range of humans and warm-blooded animals. In the current study, role of this parasite on secondary sex ratio and risk of miscarriage was investigated. In this cross-sectional study, 850 cord blood samples were collected in Tehran, Iran, 2014-2015. Enzyme-linked immunosorbent assay (ELISA) was used to assess anti-Toxoplasma IgG in samples. Information such as sex of the neonates and age, number of previous pregnancies and history of miscarriage of the mothers were recorded in questionnaires. Logistic regression analysis was used to assess the possible relationship between the latent toxoplasmosis and the highlighted parameters. Logistic regression analysis showed that the odds of having a male neonate in seropositive women is nearly 64% higher than that in seronegative women (OR = 1.64, CI95 = 1.16-2.33, P = 0.005). The odds ratio of having male neonate increased to 2.10 (CI95 = 1.24-3.57, P = 0.006) in high-titer seropositive women, compared to that in seronegative control group. The odds of having a miscarriage history was approximately two and a half times greater in seropositive women than in seronegative ones (OR = 2.45, CI95 = 1.56-3.87, P < 0.001). The odds ratio of having miscarriage increased to 2.76 (CI95 = 1.61-4.73, P < < .001) in low-titer seropositive women, compared to that in seronegative control group. Results of the current study have shown that T. gondii infection affects secondary sex ratio in human offspring and can be addressed as one of the major miscarriage causes in women.


Top-gated graphene nanoribbon transistors with ultrathin high-k dielectrics.
The integration ultrathin high dielectric constant (high-k) materials with graphene nanoribbons (GNRs) for top-gated transistors can push their performance limit for nanoscale electronics. Here we report the assembly of Si/HfO(2) core/shell nanowires on top of individual GNRs as the top-gates for GNR field-effect transistors with ultrathin high-k dielectrics. The Si/HfO(2) core/shell nanowires are synthesized by atomic layer deposition of the HfO(2) shell on highly doped silicon nanowires with a precise control of the dielectric thickness down to 1-2 nm. Using the core/shell nanowires as the top-gates, high-performance GNR transistors have been achieved with transconductance reaching 3.2 mS microm(-1), the highest value for GNR transistors reported to date. This method, for the first time, demonstrates the effective integration of ultrathin high-k dielectrics with graphene with precisely controlled thickness and quality, representing an important step toward high-performance graphene electronics.


The 'Walking for Wellbeing in the West' randomised controlled trial of a pedometer-based walking programme in combination with physical activity consultation with 12 month follow-up: rationale and study design.
Scotland has a policy aimed at increasing physical activity levels in the population, but evidence on how to achieve this is still developing. Studies that focus on encouraging real world participants to start physical activity in their settings are needed. The Walking for Well-being in the West study was designed to assess the effectiveness of a pedometer-based walking programme in combination with physical activity consultation. The study was multi-disciplinary and based in the community. Walking for Well-being in the West investigated whether Scottish men and women, who were not achieving the current physical activity recommendation, increased and maintained walking behaviour over a 12 month period. This paper outlines the rationale and design of this innovative and pragmatic study. Participants were randomised into two groups: Group 1: Intervention (pedometer-based walking programme combined with a series of physical activity consultations); Group 2: Waiting list control for 12 weeks (followed by minimal pedometer-based intervention). Physical activity (primary outcome) was measured using pedometer step counts (7 day) and the International Physical Activity Questionnaire (long version). Psychological processes were measured using questionnaires relating to the Transtheoretical Model of Behaviour Change, mood (Positive and Negative Affect Schedule) and quality of life (Euroqol EQ-5D instrument). Physiological measures included anthropometric and metabolic outcomes. Environmental influences were assessed subjectively (Neighbourhood Quality of Life Survey) and objectively (neighbourhood audit tool and GIS mapping). The qualitative evaluation employed observation, semi-structured interviews and focus groups. A supplementary study undertook an economic evaluation. Data analysis is on-going. Walking for Well-being in the West will demonstrate if a pedometer based walking programme, in combination with physical activity consultation results in a sustainable increase in walking behaviour in this sample of Scottish adults over a 12 month period. The study will examine the complex relationships between behavioural change, health consequences and the role of the environment, in conjunction with the cost effectiveness of this approach and a detailed insight into the participants' experiences of the intervention. Current Controlled Trials ISRCTN88907382.


Ultrafast Broadband Transient Absorption and Circular Dichroism Reveal Relaxation of a Chiral Copolymer.
We present a study of the photoinduced dynamics of the chiral polyfluorene-phenylene copolymer PFPh in THF and in cholesteric thin films. After photoexcitation at 370 nm in THF, ultraviolet-visible-near-infrared (UV-vis-NIR) transient absorption spectra show fast subpicosecond to picosecond intrachain migration of singlet excitons, solvation dynamics, and an exciton lifetime of 410 ps. The PFPh thin film features also interchain singlet exciton migration and exhibits shorter (2.1 and 240 ps) and longer lifetime components (2800 ps, interchain recombination). Furthermore, a setup for ultrafast UV-vis broadband transient circular dichroism (TrCD) spectroscopy has been developed. Fast supramolecular relaxation processes are observed, which are linked to changes in the anisotropic polarizability and pitch length of the cholesteric film. Such combined ultrafast transient CD and absorption experiments hold promise to reveal not only details of relaxation processes in supramolecular arrangements but also structural rearrangements of chiral molecular systems featuring CD signals in the UV-vis region.


Interplay of localized pyrene chromophores and π-conjugation in novel poly(2,7-pyrene) ladder polymers.
We present a detailed spectroscopic study, along with the synthesis, of conjugated, ladder-type 2,7-linked poly(pyrene)s. We observe a delocalization of the first singlet excited state along the polymer backbone, i.e., across the 2,7 linkage in the pyrene moiety, in contrast to earlier studies on conjugated 2,7-linked poly(pyrene)s without ladder structure. The electronic signature of the pyrene unit is, however, manifested in an increased lifetime and reduced oscillator strength as well as a modified vibronic progression in absorption of the singlet state compared to a ladder-type poly(para-phenylene) (MeLPPP). Furthermore, the reduced oscillator strength and increased lifetime slow down Förster-type energy transfer in films, where this transfer occurs to sites with increasing inter-chain coupling of H-type nature.


Retardation of autoshaping: control by contextual stimuli.
Training pigeons with random presentations of a tone and food proactively interferes with the acquisition of autoshaped keypecking to a lighted key. The interference effect is context-specific (observed only when testing for autoshaping occurs in the initial training environment). An interpretation based on blocking by background cues is suggested by the data.


Genome-wide meta-analysis identifies five new susceptibility loci for cutaneous malignant melanoma.
Thirteen common susceptibility loci have been reproducibly associated with cutaneous malignant melanoma (CMM). We report the results of an international 2-stage meta-analysis of CMM genome-wide association studies (GWAS). This meta-analysis combines 11 GWAS (5 previously unpublished) and a further three stage 2 data sets, totaling 15,990 CMM cases and 26,409 controls. Five loci not previously associated with CMM risk reached genome-wide significance (P < 5 × 10(-8)), as did 2 previously reported but unreplicated loci and all 13 established loci. Newly associated SNPs fall within putative melanocyte regulatory elements, and bioinformatic and expression quantitative trait locus (eQTL) data highlight candidate genes in the associated regions, including one involved in telomere biology.


Waste activated sludge hydrolysis and acidification: A comparison between sodium hydroxide and steel slag addition.
Alkaline treatment with steel slag and NaOH addition were investigated under different pH conditions for the fermentation of waste activated sludge. Better performance was achieved in steel slag addition scenarios for both sludge hydrolysis and acidification. More solubilization of organic matters and much production of higher VFA (volatile fatty acid) in a shorter time can be achieved at pH10 when adjusted by steel slag. Higher enzyme activities were also observed in steel slag addition scenarios under the same pH conditions. Phosphorus concentration in the supernatant increased with fermentation time and pH in NaOH addition scenarios, while in contrast most phosphorus was released and captured by steel slag simultaneously in steel slag addition scenarios. These results suggest that steel slag can be used as a substitute for NaOH in sludge alkaline treatment.


Sialoadhesin binds preferentially to cells of the granulocytic lineage.
Sialoadhesin is a macrophage-restricted, sialic acid-dependent receptor of 185 kD that binds to the oligosaccharide sequence NeuAc alpha 2,3Gal on cell surface glycoconjugates. Recent cDNA cloning has shown that sialoadhesin is a new member of the immunoglobulin superfamily with sequence similarity to CD22, a sialic acid-dependent receptor of B lymphocytes. Sialoadhesin has been implicated in cellular interactions of stromal macrophages with developing myeloid cells. In this study, direct evidence for this interaction was obtained in cell-cell binding assays using both native and recombinant forms of the protein. In all assays, sialoadhesin exhibited specific, differential binding to various murine cell populations of hemopoietic origin. In rank order, sialoadhesin bound neutrophils > bone marrow cells = blood leukocytes > lymphocytes > thymocytes. Single-cell analyses confirmed that sialoadhesin selectively bound myeloid cells in complex cell mixtures obtained from the bone marrow and blood. In comparison, a recombinant Fc-chimeric form of murine CD22 showed high binding to B and T lymphocytes, but very low binding to immature and mature myeloid cells. These results are consistent with the notion that sialoadhesin in involved in interactions with granulocytes at different stages of their life histories.


Sr31-Virulent Races (TTKSK, TTKST, and TTTSK) of the Wheat Stem Rust Pathogen Puccinia graminis f. sp. tritici are Present in Tanzania.
Since the first detection of race TTKSK (syn. Ug99) in Uganda in 1999 (2), the migration and evolution of Sr31-virulent races of Puccinia graminis f. sp. tritici [Pgt] have been closely monitored, particularly in Kenya and countries north, along the likely trajectory of migration to major wheat-producing regions of Asia. More recently, surveillance efforts have been undertaken to the south as well, and Ug99-related races have been detected in South Africa and Zimbabwe (3,4). Here we report for the first time results of a survey conducted in Tanzania. Systematic race surveillance provides data not only on the current distribution of the Ug99 race group, but also on the possible points of origin as well as the pace and probable paths of dispersal of future races from the region. In this context, the presence or absence of the Ug99 group of wheat stem rust races in adjacent countries like Tanzania assumes regional, and possibly global, relevance. A preliminary survey conducted in September 2006 indicated the presence of Sr31-virulent races of Pgt outside Slahhamo Village (3°15'S, 35°48'E) in the Ngorongoro highlands of northern Tanzania, based on compatible reactions with cv. K-Mamba (a.k.a. Mwamba), a cultivar whose pedigree indicates the presence of Sr31. A broader survey was conducted in August 2009, during which infected tissue was collected from currently-grown cultivars in research plots and on large estates, as well as from the mixes of older cultivars common on smallholder farms. In all, Pgt-infected samples were collected from one site in the Arusha region [Monduli (3°16' S, 36°24'E)], three sites in the Ngorongoro highlands [Karatu (3°20' S, 35°40' E), Upper Kitete (3°14' S, 35°53' E), and Slahhamo], one site in the Manyara region [Hanang (4°43' S, 35°40' E)], and one site in the Mbeya region [southern highlands (8°87' S, 33°40' E)], thereby giving representation to all four major wheat growing areas in the country. Sample storage, inoculation, incubation, disease assessment, and derivation of single-pustule cultures were all performed according to the methods described by Jin et al. (1). In addition to the 20 differentials in the expanded Pgt differential set of North America, we included two supplemental tester lines: Siouxland (Sr24 + Sr31) and Sisson (Sr31 + Sr36). Each single-pustule-derived isolate was evaluated for virulence on the differential and supplemental lines at least twice. A total of 39 single-pustule isolates were derived from the six collection sites. All 39 isolates were identified as belonging to the Ug99 race group, with six identified as TTKSK (all four regions), 30 identified as TTKST (Sr31 + Sr24 virulence; Arusha region and the Ngorongoro highlands), and three identified as TTTSK (Sr31 + Sr36 virulence; Manyara region and the Ngorongoro highlands). The results of this study suggest that, to more precisely locate the "hot spots" and thereby gain a better understanding of the mechanisms of novel race emergence in East Africa, it would be prudent to include Tanzania, heretofore a blank area on the wheat rust surveillance map, in future systematic race monitoring efforts. References: (1) Y. Jin et al. Plant Dis. 92:923, 2008. (2) Z. A. Pretorius et al. Plant Dis. 84:203, 2000. (3) Z. A. Pretorius et al. Plant Dis. 94:784, 2010. (4) Z. A. Pretorius et al. Plant Dis. 96:590, 2012.


Cytoplasmic type 80S ribosomes associated with yeast mitochondria. IV. Attachment of ribosomes to the outer membrane of isolated mitochondria.
Growing yeast spheroplasts were shown to have, on the average, four times the number of cytoplasmic ribosomes in contact with the outer mitochondrial membrane compared to starved spheroplasts. Ribosomes in contact with mitochondria in the growing spheroplast preparation, like free cytoplasmic ribosomes, exist primarily as polysome structures. In the starved spheroplast preparation, both mitochondria-bound and free cytoplasmic ribosomes exist primarily as monosomes. Mitochondria isolated from growing spheroplasts in a medium containing lmM Mg++ have cytoplasmic ribosomes bound directly to the outer membrane. These ribosomes can be quantitatively removed by washing the mitochondria with 2 mM EDTA. Mitochondria from starved spheroplasts are capable of accepting either free cytoplasmic polysomes or cytoplasmic polysomes extracted from mitochondria. However, the extent of polysome binding to mitochondria was shown to be a direct function of the Mg++ concentration; a smaller percentage of the input polysomes bind as the Mg++ concentration is lowered. At 1 mM Mg++, neither free cytoplasmic nor mitochondria-bound polysomes bind to mitochondria. Nevertheless, when growing spheroplasts are broken and mitochondria isolated in medium containing 1 mM Mg++, the mitochondria are seen to have cytoplasmic ribosomes firmly attached to the outer membrane. This result, in addition to our earlier data (Kellems, R. E., and R. A. Butow. 1974. J. Biol. Chem. 249:3304-3310), support the view that cytoplasmic ribosomes attached to the outer membrane of purified mitochondria were attached in vivo. In preparations of mitochondria isolated from growing spheroplasts, ribosomes appear to be found to specific regions of the outer membrane, namely those regions which are in close association or in contact with the inner mitochondrial membrane. This is particularly evident with mitochondria in a condensed configuration. This finding suggests a mechanism whereby cytoplasmically synthesized mitochondrial protein could be transferred by a process of vectorial translation across both membranes of the organelle.


Regulation of hippocampal cannabinoid CB1 receptor actions by adenosine A1 receptors and chronic caffeine administration: implications for the effects of Δ9-tetrahydrocannabinol on spatial memory.
The cannabinoid CB(1) receptor-mediated modulation of γ-aminobutyric acid (GABA) release from inhibitory interneurons is important for the integrity of hippocampal-dependent spatial memory. Although adenosine A(1) receptors have a central role in fine-tuning excitatory transmission in the hippocampus, A(1) receptors localized in GABAergic cells do not directly influence GABA release. CB(1) and A(1) receptors are the main targets for the effects of two of the most heavily consumed psychoactive substances worldwide: Δ(9)-tetrahydrocannabinol (THC, a CB(1) receptor agonist) and caffeine (an adenosine receptor antagonist). We first tested the hypothesis that an A(1)-CB(1) interaction influences GABA and glutamate release in the hippocampus. We found that A(1) receptor activation attenuated the CB(1)-mediated inhibition of GABA and glutamate release and this interaction was manifested at the level of G-protein activation. Using in vivo and in vitro approaches, we then investigated the functional implications of the adenosine-cannabinoid interplay that may arise following chronic caffeine consumption. Chronic administration of caffeine in mice (intraperitoneally, 3 mg/kg/day, for 15 days, >12 h before trials) led to an A(1)-mediated enhancement of the CB(1)-dependent acute disruptive effects of THC on a short-term spatial memory task, despite inducing a reduction in cortical and hippocampal CB(1) receptor number and an attenuation of CB(1) coupling with G protein. A(1) receptor levels were increased following chronic caffeine administration. This study shows that A(1) receptors exert a negative modulatory effect on CB(1)-mediated inhibition of GABA and glutamate release, and provides the first evidence of chronic caffeine-induced alterations on the cannabinoid system in the cortex and hippocampus, with functional implications in spatial memory.


The role of family physicians in increasing annual fecal occult blood test screening coverage: a prospective intervention study.
Colorectal cancer is the second leading cause of cancer mortality in israel. Unfortunately, compliance with annual fecal occult blood testing is very low. To assess the effectiveness of interventions to increase FOBT screening in primary care clinics in Israel. A prospective, randomized study included all 50-75 year old enrollees of six family physicians in two primary care clinics. The register of two physicians, one from each clinic, was allocated to one of three groups. Two FOBT reminder strategies were tested: a physician reminder (753 patients), and a patient reminder that was either a phone call (312 patients) or a letter (337 patients). The control group (913 patients) of physicians continued administering their regular level of care. The main outcome measure was the percentage of patients undergoing FOBT screening in each study arm at the conclusion of the one year study period. In the intervention groups 14.3% (201/1,402) were screened using the FOBT over the course of the study year. Using an intent-to-screen analysis, the screening rate in the physician and patient reminder groups was significantly higher than in the control group (16.5 and 11.9%, vs. 1.2% respectively, P < 0.0001). Phone reminders were significantly more effective than letters (14.7 vs. 9.2%, P = 0.01). Various reminder systems for FOBT are beneficial, especially those centered around the family physician. Further research should focus on this area, in conjunction with other novel approaches.


Autophagy modulates Aβ accumulation and formation of aggregates in yeast.
Intracellular accumulation of amyloid-β protein (Aβ) is an early event in Alzheimer's disease (AD). The autophagy-lysosomal pathway is an important pathway for maintaining cellular proteostasis and for the removal of damaged organelles and protein aggregates in all eukaryotes. Despite mounting evidence showing that modulating autophagy promotes clearance of Aβ aggregates, the regulatory mechanisms and signalling pathways underlying this process remain poorly understood. In order to gain better insight we used our previously characterised yeast model expressing GFP-Aβ42 to identify genes that regulate the removal of Aβ42 aggregates by autophagy. We report that GFP-Aβ42 is sequestered and is selectively transported to vacuole for degradation and that autophagy is the prominent pathway for clearance of aggregates. Next, to identify genes that selectively promote the removal of Aβ42 aggregates, we screened levels of GFP-Aβ42 and non-aggregating GFP-Aβ42 (19:34) proteins in a panel of 192 autophagy mutants lacking genes involved in regulation and initiation of the pathway, cargo selection and degradation processes. The nutrient and stress signalling genes RRD1, SNF4, GCN4 and SSE1 were identified. Deletion of these genes impaired GFP-Aβ42 clearance and their overexpression reduced GFP-Aβ42 levels in yeast. Overall, our findings identify a novel role for these nutrient and stress signalling genes in the targeted elimination of Aβ42 aggregates, which offer a promising avenue for developing autophagy based therapies to suppress amyloid deposition in AD.


The genetic basis of adaptive population differentiation: a quantitative trait locus analysis of fitness traits in two wild barley populations from contrasting habitats.
We used a quantitative trait locus (QTL) approach to study the genetic basis of population differentiation in wild barley, Hordeum spontaneum. Several ecotypes are recognized in this model species, and population genetic studies and reciprocal transplant experiments have indicated the role of local adaptation in shaping population differences. We derived a mapping population from a cross between a coastal Mediterranean population and a steppe inland population from Israel and assessed F3 progeny fitness in the natural growing environments of the two parental populations. Dilution of the local gene pool, estimated as the proportion of native alleles at 96 marker loci in the recombinant lines, negatively affected fitness traits at both sites. QTLs for fitness traits tended to differ in the magnitude but not in the direction of their effects across sites, with beneficial alleles generally conferring a greater fitness advantage at their native site. Several QTLs showed fitness effects at one site only, but no opposite selection on individual QTLs was observed across the sites. In a common-garden experiment, we explored the hypothesis that the two populations have adapted to divergent nutrient availabilities. In the different nutrient environments of this experiment, but not under field conditions, fitness of the F3 progeny lines increased with the number of heterozygous marker loci. Comparison of QTL-effects that underlie genotype x nutrient interaction in the common-garden experiment and genotype x site interaction in the field suggested that population differentiation at the field sites may have been driven by divergent nutrient availabilities to a limited extent. Also in this experiment no QTLs were observed with opposite fitness effects in contrasting environments. Our data are consistent with the view that adaptive differentiation can be based on selection on multiple traits changing gradually along ecological gradients. This can occur without QTLs showing opposite fitness effects in the different environments, that is, in the absence of genetic trade-offs in performance between environments.


Probabilistic sensitivity analysis in cost-effectiveness. An application from a study of vaccination against pneumococcal bacteremia in the elderly.
We explore the policy implications of probabilistic sensitivity analysis in cost-effectiveness analysis by applying simulation methods to a decision model. We present the multiway sensitivity analysis results of a study of the cost-effectiveness of vaccination against pneumococcal bacteremia in the elderly. We then execute a probabilistic sensitivity analysis of the cost-effectiveness ratio by specifying posterior distributions for the uncertain parameters in our decision analysis model. In order to estimate probability intervals, we rank the numerical values of the simulated incremental cost-effectiveness ratios (ICERs) to take into account preferences along the cost-effectiveness plane. The 95% probability intervals for the ICER were generally much narrower than the difference between the best case and worst case results from a multiway sensitivity analysis. Although the multiway sensitivity analysis had indicated that, in the worst case, vaccination in the 85 and older age group was not acceptable from a policy standpoint, probabilistic methods indicated that the cost-effectiveness of vaccination was below $50,000 per quality-adjusted life-year in greater than 92% of the simulations and below $100,000 in greater than 95% of the simulations. Probabilistic methods can supplement multiway sensitivity analyses to provide a more comprehensive picture of the uncertainty associated with cost-effectiveness ratios and thereby inform policy decisions.


Global Overview of the Epidemiology of Atherosclerotic Cardiovascular Disease.
Atherosclerotic cardiovascular disease (ACD) is the leading cause of mortality worldwide. The objective of this paper is to provide an overview of the global burden of ACD and its risk factors and to discuss the main challenges and opportunities for prevention. Publicly available data from the Global Burden of Disease Study were analyzed for ischemic heart disease (IHD), ischemic stroke and ACD risk factors. Data from the WHO Global Health Observatory were used to describe prevalence of diverse cardiometabolic risk factors. World Bank Gross Domestic Product per capita (GDPc) information was used to categorize countries according to income level. Cardiovascular mortality decreased globally from 1990-2010 with important differences by GDPc; during 1990 there was a positive association between IHD mortality and GDPc. Higher-income countries had higher rates compared to those of lower-income countries. High levels of body mass index (BMI), blood pressure, glucose and cholesterol have a differential contribution to mortality by income group over time; high-income countries have been able to reduce the contribution from these risk factors in the last 20 years, whereas lower/middle income countries show an increasing trend in mortality attributable to high BMI and glucose. Although age-adjusted ACD mortality rate trends decreased globally, the absolute number of ACD deaths is increasing in part due to the growth of the population and aging, as well as to important lifestyle and food-system changes that likely attenuate gains in prevention. Population and individual level preventable causes of ACD must be aggressively and efficiently targeted in countries of lower economic development in order to reduce the growing burden of disease due to ACD.


Body mass index and its effect on outcome in children after lung transplantation.
Malnutrition is common in children undergoing lung transplantation, particularly among those with cystic fibrosis (CF). However, the effect of body habitus on outcome after pediatric lung transplantation is unknown. We studied body mass index (BMI) and its effect on outcome in pediatric lung transplantation. The International Society for Heart and Lung Transplantation Registry on Pediatric Lung Transplantation was queried for primary pediatric lung transplant recipients (aged<18 years) between 1990 and 2008. BMI cohorts were defined according to International Obesity Task Force cutoffs: thinness grade 3, BMI<16 kg/m(2); thinness grade 2, 16 to<17 kg/m(2); thinness grade 1, 17 to<18.5 kg/m(2); normal, 18.5 to<25 kg/m(2); overweight, 25 to<30 kg/m(2); and obese, ≥ 30 kg/m(2). Survival was compared among BMI cohorts within CF and non-CF recipient groups. Included were 897 recipients. The median age at transplantation was 14 years (interquartile, 11, 16 years) and 63% had CF. The incidence of thinness was 59% in CF vs 39% in non-CF patients (p<0.001). A significant proportion of CF patients were underweight, whereas more non-CF patients were obese. Cox regression showed neither underweight nor overweight CF recipients differed in survival compared with recipients of normal-weight recipients. Grade of thinness was not related to outcome after transplantation. For non-CF recipients, being overweight/obese increased risk of death compared with normal-weight recipients (hazard ratio, 2.05; 95% confidence interval, 1.28-3.26; p = 0.002). The incidence of underweight status amongst pediatric lung transplant recipients with CF is high. However, we did not find a significant negative effect of underweight body habitus on survival in CF children after lung transplantation. Overweight pediatric recipients appear to have poorer survival after transplant.


Blood-based cerebral biomarkers in preeclampsia: Plasma concentrations of NfL, tau, S100B and NSE during pregnancy in women who later develop preeclampsia - A nested case control study.
To evaluate if concentrations of the neuronal proteins neurofilament light chain and tau are changed in women developing preeclampsia and to evaluate the ability of a combination of neurofilament light chain, tau, S100B and neuron specific enolase in identifying neurologic impairment before diagnosis of preeclampsia. A nested case-control study within a longitudinal study cohort was performed. 469 healthy pregnant women were enrolled between 2004-2007 and plasma samples were collected at gestational weeks 10, 25, 28, 33 and 37. Plasma concentrations of tau and neurofilament light chain were analyzed in 16 women who eventually developed preeclampsia and 36 controls throughout pregnancy with single molecule array (Simoa) method and compared within and between groups. S100B and NSE had been analyzed previously in the same study population. A statistical model with receiving characteristic operation curve was constructed with the four biomarkers combined. Plasma concentrations of neurofilament light chain were significantly increased in women who developed preeclampsia in gestational week 33 (11.85 ng/L, IQR 7.48-39.93 vs 6.80 ng/L, IQR 5.65-11.40) and 37 (22.15 ng/L, IQR 10.93-35.30 vs 8.40 ng/L, IQR 6.40-14.30) and for tau in gestational week 37 (4.33 ng/L, IQR 3.97-12.83 vs 3.77 ng/L, IQR 1.91-5.25) in contrast to healthy controls. A combined model for preeclampsia with tau, neurofilament light chain, S100B and neuron specific enolase in gestational week 25 displayed an area under the curve of 0.77, in week 28 it was 0.75, in week 33 it was 0.89 and in week 37 it was 0.83. Median week for diagnosis of preeclampsia was at 38 weeks of gestation. Concentrations of both tau and neurofilament light chain are increased in the end of pregnancy in women developing preeclampsia in contrast to healthy pregnancies. Cerebral biomarkers might reflect cerebral involvement before onset of disease.


Deletions of distal 9p associated with 46,XY male to female sex reversal: definition of the breakpoints at 9p23.3-p24.1.
Monosomy of distal 9p is associated in rare cases with abnormalities of testicular determination, which can lead to male to female sex reversal in a 46,XY genetic background. We present two 46,XY individuals partially monosomic for 9p who were raised as females. Definition of the breakpoints using somatic cell hybrids containing only the rearranged chromosome 9 indicated that in the first patient the breakpoint was located between markers D9S256 and D9S144 and in the second patient, the breakpoint was distal to the marker D9S144. In both cases this corresponds to the cytogenetic position 9p23.3-p24.1. Analysis of highly polymorphic microsatellite markers demonstrated a paternal origin of the rearranged chromosome 9 in both patients. These studies define the minimum region associated with male to female sex reversal as 9p24.1-pter.


Enzymatic synthesis using glycoside phosphorylases.
Carbohydrate phosphorylases are readily accessible but under-explored catalysts for glycoside synthesis. Their use of accessible and relatively stable sugar phosphates as donor substrates underlies their potential. A wide range of these enzymes has been reported of late, displaying a range of preferences for sugar donors, acceptors and glycosidic linkages. This has allowed this class of enzymes to be used in the synthesis of diverse carbohydrate structures, including at the industrial scale. As more phosphorylase enzymes are discovered, access to further difficult to synthesise glycosides will be enabled. Herein we review reported phosphorylase enzymes and the glycoside products that they have been used to synthesise.


Atypical antibody responses to influenza.
Influenza viruses undergo rapid antigenic evolution and reassortment, resulting in annual epidemics and the occasional pandemics. Exposure to influenza virus hemagglutinin (HA) and neuraminidase (NA) antigen, either through vaccination or infection, induces an antibody response able to recognize only the homologous antigenic subtype. However, atypical antibody responses recognizing non-homologous influenza subtypes have been reported during infection and vaccination. Here, we review the incidence of these phenomena in published literature and discuss the potential mechanisms underlying them.


Pharmacokinetics, pharmacodynamics and pharmacogenetics associated with nonsteroidal anti-inflammatory drugs and opioids in pediatric cancer patients.
Advancing appropriate and adequate analgesic pharmacotherapy in pediatric patients with cancer is an area of clinical need. Few studies have been performed to evaluate the selection of an analgesic and appropriate dosing corresponding to analgesic effect among pediatric cancer patients. This review describes information related to pharmacokinetic, pharmacodynamic, and pharmacogenomic (when applicable) considerations for analgesics that are commonly used to manage pain experienced by pediatric patients with cancer. Areas covered: Analgesics commonly used to treat pediatric patients with malignancy patterned after the World Health Organization's 'analgesic ladder' for cancer pain management. Expert opinion: Addressing pain management safely and effectively in pediatric patients with cancer will require advances in both drug development, to increase the armament of analgesics available for children, and our pharmacologic understanding of those analgesics in current use. However, performing the necessary types of studies to develop new analgesics, or gain knowledge of existing therapy, within a population that is relatively small, diverse, and who experience pain originating from a variety of sources, is a tremendous challenge.


Cytokine Profiles Associated With Worse Prognosis in a Hospitalized Peruvian COVID-19 Cohort.
Cytokines, chemokines and growth factors present different expression profiles related to the prognosis of COVID-19. We analyzed clinical parameters and assessed the expression of these biomarkers in patients with different disease severity in a hospitalized Peruvian cohort to determine those associated with worse prognosis. We measured anti-spike IgG antibodies by ELISA and 30 cytokines by quantitative suspension array technology in 123 sera samples. We analyzed differences between patients with moderate, severe and fatal COVID-19 by logistic regression at baseline and in longitudinal samples. Significant differences were found among the clinical parameters: hemoglobin, neutrophils, lymphocytes and C-reactive protein (CRP), creatinine and D-dimer levels. Higher anti-spike IgG antibody concentrations were associated to fatal patient outcomes. At hospitalization, IL-10, IL-6, MIP-1α, GM-CSF, MCP-1, IL-15, IL-5, IL1RA, TNFα and IL-8 levels were already increased in fatal patients´ group. Meanwhile, multivariable analysis revealed that increased GM-CSF, MCP-1, IL-15, and IL-8 values were associated with fatal outcomes. Moreover, longitudinal analysis identified IL-6 and MCP-1 as the main risk factors related to mortality in hospitalized COVID-19 patients. In this Peruvian cohort we identified and validated biomarkers related to COVID-19 outcomes. Further studies are needed to identify novel criteria for stratification of SARS-CoV-2 infected patients at hospital entry. In the most severe forms of SARS-CoV-2 infection, large numbers of innate and adaptive immune cells become activated and begin to produce pro-inflammatory cytokines, establishing an exacerbated feedback loop of inflammation. A total of 55 patients with laboratory-confirmed COVID-19 admitted to the Hospital Nacional Guillermo Almenara Irigoyen in Lima, Peru were enrolled during August-October 2020. Of these, 21 had moderate disease, 24 severe diseases and 10 died. We measured 30 cytokines and chemokines by quantitative suspension array technology and anti-spike IgG antibodies using a commercial ELISA. We evaluated these parameters in peripheral blood every 2-5 days until patient discharge or death. Patient information and clinical parameters related were obtained from the respective clinical histories. The frequency of obesity differed among the 3 groups, being most frequent in patients who died. There were also significant differences in clinical parameters: hemoglobin, segmented neutrophils, lymphocytes,C-reactive protein, creatinine and D-dimer levels. Greater anti-spike IgG antibody concentrations were associated to fatal outcomes. In univariate analyses, higher baseline concentrations of IL-6, MIP-1α, GM-CSF, MCP-1, IL-15, IL-5, IL1RA, TNFα, IL-8 and IL-12p70 correlated with severity, while multivariable analysis showed that increased concentrations in 4 biomarkers (GM-CSF, MCP-1, IL-15, IL-8) were associated with fatal outcomes. Longitudinal analysis showed IL-6 (hazard ratio [HR] 6.81, 95% confidence interval [CI] 1.6-28.7) and MCP-1 (HR 4.61, 95%CI 1.1-19.1) to be related to mortality in hospitalized COVID-19 patients. Cytokine, chemokine and growth factor profiles were identified and validated related to severity and outcomes of COVID-19. Our findings may be useful to identify novel criteria for COVID-19 patient stratification at hospital entry.


An under-recognized influenza epidemic identified by rapid influenza testing, southern Sri Lanka, 2013.
Influenza accounts for a large burden of acute respiratory tract infections in high-income countries; data from lower-income settings are limited due to lack of confirmatory testing. Consecutive outpatients presenting to the largest tertiary care hospital in southern Sri Lanka were surveyed for influenza-like illness (ILI), defined as acute onset of fever ≥ 38.0°C and cough. Patients were administered a questionnaire and nasal/nasopharyngeal sampling for rapid influenza A/B testing. We enrolled 311 patients with ILI from March to November 2013: 170 (54.7%) children and 172 (55.3%) males. Approximately half (147, 47.3%) tested positive for influenza, but 253 (81.4%) were prescribed antibiotics. On bivariable analysis, symptoms associated with influenza included pain with breathing (P < 0.001), headache (P = 0.005), fatigue (P = 0.003), arthralgias (P = 0.003), and myalgias (P = 0.006) in children and pain with breathing (P = 0.01), vomiting (P = 0.03), and arthralgias (P = 0.03) in adults. Our final clinical predictive models had low sensitivity and fair specificity-50.0% (95% CI: 38.6-61.4%) and 83.2% (95% CI: 73.4-90.0%), respectively, in children and 52.2% (95% CI: 39.9-64.2%) and 81.4% (95% CI: 70.0-89.4%), respectively, in adults. Our study confirms the ability of rapid influenza testing to identify an influenza epidemic in a setting in which testing is not routinely available.


The genome of the sponge Amphimedon queenslandica provides new perspectives into the origin of Toll-like and interleukin 1 receptor pathways.
Members of the Toll-like receptor (TLR) and the interleukin 1 receptor (IL1R) superfamilies activate various signaling cascades that are evolutionarily conserved in eumetazoans. In this study, we have searched the genome and expressed sequence tags of the demosponge Amphimedon queenslandica for molecules involved in TLR and IL1R signaling. Although we did not identify a conventional TLR or ILR, the Amphimedon genome encodes two related receptors, AmqIgTIRs, which are comprised of at least three extracellular IL1R-like immunoglobulins (Ig) and an intracellular TLR-like Toll/interleukin1 receptor/resistance (TIR) domain. The remainder of the TLR/IL1R pathway is mostly conserved in Amphimedon and includes genes known to interact with TLRs and IL1Rs in bilaterians, such as Toll-interacting protein (Tollip) and myeloid differentiation factor 88 (MyD88). By comparing the sponge genome to that of nonmetazoan eukaryotes and other basal animal phyla (i.e., placozoan and cnidarian representatives) we can infer that most components of the signaling cascade, including the receptors, evolved after the divergence of metazoan, and choanoflagellate lineages. In most cases, these proteins are composed of metazoan-specific domains (e.g., Pellino) or architectures (e.g., the association of a death domain with a TIR domain in the MyD88). The dynamic expression of the two AmqIgTIRs, AmqMyD88, AmqTollip, and AmqPellino during Amphimedon embryogenesis and larval development is consistent with the TLR/IL1R pathway having a role in both development and immunity in the last common metazoan ancestor.


Development and Validation of a Nomogram for Predicting Survival in Patients with Thyroid Cancer.
BACKGROUND The AJCC staging system is inadequate for use in patients with thyroid carcinomas. Here, we aimed to establish a nomogram for thyroid cancer, and we compare its prognostic value with the AJCC staging system in adults diagnosed with thyroid carcinoma. MATERIAL AND METHODS Patient records were obtained from the Surveillance, Epidemiology, and End Result database. The 8491 included patients were divided into a modeling cohort (n=5943) and a validation cohort (n=2548). The variables included in the modeling cohort were selected using a backward stepwise selection method with Cox regression, and the prognosis nomogram was constructed. In the validation cohort, we compared our survival model with the AJCC prognosis model using the concordance index, the area under the time-dependent receiver operating characteristic curve, the net reclassification improvement, the integrated discrimination improvement, calibration plotting, and decision curve analysis. RESULTS Twelve independent prognostic factors were identified and used to establish the nomogram. In particular, marital status was included in a survival prediction model of thyroid cancer for the first time. The concordance index, area under the time-dependent receiver operating characteristic curve, net reclassification improvement, integrated discrimination improvement, calibration plotting, and decision curve analysis for the nomogram showed better performance compared to the AJCC staging system. CONCLUSIONS We have developed and validated a highly accurate thyroid cancer prognosis nomogram. The prognostic value of the nomogram is better than that of the AJCC staging system alone.


Lysophospholipids induce membrane hyperpolarization in microglia by activation of IKCa1 Ca(2+)-dependent K(+) channels.
Effects of the lysophospholipids sphingosine-1-phosphate and lysophosphatidic acid were studied in cultured murine microglia using the patch-clamp and video imaging techniques. Both lysophospholipids induced transient membrane hyperpolarization and K(+) current activation. The lysophospholipid-induced K(+) current was blocked by charybdotoxin or iberiotoxin, but was unaffected by apamin. In recordings with 1 microM intracellular free Ca(2+), Ca(2+)-dependent K(+) currents of microglia showed a similar pharmacological profile to lysophospholipid-induced currents. The Ca(2+)-dependent K(+) channels activated in microglia by lysophospholipids are most likely encoded by the IKCa1 channel gene. The presence of IKCa1 mRNA in microglia was demonstrated by reverse transcriptase-polymerase chain reaction studies. Ca(2+) imaging experiments revealed increases in the intracellular free Ca(2+) concentration of microglia to a mean value of about 400 nM after application of 1 microM sphingosine-1-phosphate or 1 microM lysophosphatidic acid. We suggest that the transient membrane hyperpolarization seen in microglia following exposure to sphingosine-1-phosphate or lysophosphatidic acid is caused by activation of IKCa1 Ca(2+)-dependent K(+) channels. Increases in the concentration of intracellular free Ca(2+) evoked by the lysophospholipids are sufficient to activate microglial Ca(2+)-dependent K(+) channels.


Pyrrolo[2,3-d]pyrimidines and pyrido[2,3-d]pyrimidines as conformationally restricted analogues of the antibacterial agent trimethoprim.
Conformationally restricted analogues of the antibacterial agent trimethoprim (TMP) were designed to mimic the conformation of drug observed in its complex with bacterial dihydrofolate reductase (DHFR). This conformation of TMP was achieved by linking the 4-amino function to the methylene group by one- and two-carbon bridges. A pyrrolo[2,3-d]pyrimidine, a dihydro analogue, and a tetrahydropyrido[2,3-d]pyrimidine were synthesized and tested as inhibitors of DHFR. One analogue showed activity equivalent to that of TMP against DHFR from three species of bacteria. An X-ray crystal structure of this inhibitor bound to Escherichia coli DHFR was determined to evaluate the structural consequences of the conformational restriction.


Correlation of the glycoside response, the force staircase, and the action potential configuration in the neonatal rat heart.
The rat heart demonstrates marked alterations in its responses to ouabain and increased frequencies of stimulation and in the duration of its action potential during the initial 21 days of life. At an age of 6.2 days 5 times 10- minus 5M ouabain produced a 158.2% increase in dP/dt compared with a 17.2% increase at 21.1 days (P less than 0.001). At 6.2 days dP/dt increased by 53.4% when the heart rate was accelerated from 30 to 90 beats/min compared with an increase of 12.2% at 21.1 days (P less than 0.005). The positive glycoside and staircase responses at the younger age were virtually eliminated when the hearts were perfused with a solution containing 50% [Na+]o and 25% [Ca-2+]o ([Ca-2+]o/[Na+]o-2 maintained constant). The duration of the ventricular action potential progressively decreased from 350-400 msec at birth to 100-150 msec at 21 days of life. This decrease was due to a shortening and a decrease in the potential level of the plateau phase. The prominent plateau typical of the early neonatal period was significantly diminished by perfusion with 50% [Na+]o. The results suggest that Na+ flux through a slow membrane channel plays a significant role in the positive staircase and glycoside responses of the early neonatal rat heart. As the heart matures and becomes functionally anomalous relative to other mammalian species, the slow channel progressively closes.


Conversion of normal human oral keratinocytes to tumorigenic cells is associated with the acquisition of resistance to TGF-beta.
Normal human epithelial cells cannot proliferate and undergo apoptosis in the presence of transforming growth factor-beta (TGF-beta) in vitro, but many human epidermoid cancer cells are resistant to TGF-beta. Resistance to TGF-beta may thus, in part, be responsible for uncontrolled proliferation of cancer cells. Though detailed mechanisms for the resistance of cancer cells to TGF-beta remain unknown, resistance may be due to decreased expression of TGF-beta receptors from cancer cells. To investigate this possibility, we determined the expression of TGF-beta and type II TGF-beta receptor in primary normal human oral keratinocytes (NHOK), human papillomavirus-immortalized human oral keratinocytes (HOK-16B) and two tumor cell lines derived from HOK-16B (CTHOK-16B-BaP and CTHOK-16B-DMBA). Our results show that (1) the cellular and secretory TGF-beta levels in immortalized and tumor cells were notably lower than in NHOK and (2) the level of type II TGF-beta receptor of the tested cells was similar to each other. Taken together, the conversion of NHOK to tumorigenic cells may, in part, be due to the acquisition of NHOK resistance to TGF-beta through underexpression of this cytokine.


Amniotic membrane-derived mesenchymal cells and their conditioned media: potential candidates for uterine regenerative therapy in the horse.
Amniotic membrane-derived mesenchymal cells (AMCs) are considered suitable candidates for a variety of cell-based applications. In view of cell therapy application in uterine pathologies, we studied AMCs in comparison to cells isolated from the endometrium of mares at diestrus (EDCs) being the endometrium during diestrus and early pregnancy similar from a hormonal standpoint. In particular, we demonstrated that amnion tissue fragments (AM) shares the same transcriptional profile with endometrial tissue fragments (ED), expressing genes involved in early pregnancy (AbdB-like Hoxa genes), pre-implantation conceptus development (Erα, PR, PGRMC1 and mPR) and their regulators (Wnt7a, Wnt4a). Soon after the isolation, only AMCs express Wnt4a and Wnt7a. Interestingly, the expression levels of prostaglandin-endoperoxide synthase 2 (PTGS2) were found greater in AM and AMCs than their endometrial counterparts thus confirming the role of AMCs as mediators of inflammation. The expression of nuclear progesterone receptor (PR), membrane-bound intracellular progesterone receptor component 1 (PGRMC1) and membrane-bound intracellular progesterone receptor (mPR), known to lead to improved endometrial receptivity, was maintained in AMCs over 5 passages in vitro when the media was supplemented with progesterone. To further explore the potential of AMCs in endometrial regeneration, their capacity to support resident cell proliferation was assessed by co-culturing them with EDCs in a transwell system or culturing in the presence of AMC-conditioned medium (AMC-CM). A significant increase in EDC proliferation rate exhibited the crucial role of soluble factors as mediators of stem cells action. The present investigation revealed that AMCs, as well as their derived conditioned media, have the potential to improve endometrial cell replenishment when low proliferation is associated to pregnancy failure. These findings make AMCs suitable candidates for the treatment of endometrosis in mares.


Time to pregnancy: a computational method for using the duration of non-conception for predicting conception.
An important problem in reproductive medicine is deciding when people who have failed to become pregnant without medical assistance should begin investigation and treatment. This study describes a computational approach to determining what can be deduced about a couple's future chances of pregnancy from the number of menstrual cycles over which they have been trying to conceive. The starting point is that a couple's fertility is inherently uncertain. This uncertainty is modelled as a probability distribution for the chance of conceiving in each menstrual cycle. We have developed a general numerical computational method, which uses Bayes' theorem to generate a posterior distribution for a couple's chance of conceiving in each cycle, conditional on the number of previous cycles of attempted conception. When various metrics of a couple's expected chances of pregnancy were computed as a function of the number of cycles over which they had been trying to conceive, we found good fits to observed data on time to pregnancy for different populations. The commonly-used standard of 12 cycles of non-conception as an indicator of subfertility was found to be reasonably robust, though a larger or smaller number of cycles may be more appropriate depending on the population from which a couple is drawn and the precise subfertility metric which is most relevant, for example the probability of conception in the next cycle or the next 12 cycles. We have also applied our computational method to model the impact of female reproductive ageing. Results indicate that, for women over the age of 35, it may be appropriate to start investigation and treatment more quickly than for younger women. Ignoring reproductive decline during the period of attempted conception added up to two cycles to the computed number of cycles before reaching a metric of subfertility.


Precursors of CD3+CD4+CD8+ cells in the human thymus are defined by expression of CD34. Delineation of early events in human thymic development.
Studies of the most immature T cell progenitors in the human thymus have been hampered by the lack of markers and assays that define these cells. In this report we used a novel human fetal thymic organ culture system to determine the potential of T cell precursors isolated from human postnatal thymus, to differentiate into CD3+ thymocytes, and to investigate early stages of human T cell development. It was found that thymocytes that lack the markers CD3, CD4, and CD8 (triple negative [TN]) can differentiate in an allogeneic organotypic thymic culture. The capacity of TN thymocytes to differentiate was exclusively confined to the CD34+ population. CD34- TN thymocytes failed to differentiate in this system. In contrast, cloned lines of CD3- thymocytes could only be established from CD34- TN thymocytes. Five subsets of CD3- thymocytes were found with the following phenotype: CD1-TN, CD1+TN, CD1+CD4+CD8-, CD1+CD4+CD8 alpha+ beta-, and CD1+CD4+CD8 alpha beta+. These subpopulations expressed decreasing levels of CD34. The CD1-CD3- population expressed the highest levels of CD34 supporting the notion that this population is the most immature T cell precursor in the thymus, whereas the CD1+CD4+CD8 alpha+ beta+ which did not express CD34 seems to be the most mature of these CD3- populations. This notion is supported by the observations that CD34+ cells isolated from fetal liver, which differentiated into T cells in a FTOC, developed into CD3+ cells via CD1- and CD4+CD8- intermediates. Based on these data, we present a model of early stages in human intrathymic development.


Illusions of necessity: evading responsibility for choice in health care.
Discretionary power is commonly defended by denial of its existence, the allegation of inevitability. Objective external conditions and forces are claimed to dictate policy decisions with tangible distributional effects. In health policy, such forces include the aging of the population, the extension of technology, and the demands of ethical standards. Taken together, these forces create relentless upward pressure on costs, to levels which society "cannot afford," necessitating sacrifice of the interests of the "less eligible." Yet quantitative analysis of these forces does not sustain the argument; in each case the source of cost escalation is not external pressure but the way in which the health care system itself reacts. Less costly and equally effective options are demonstrably available, but would threaten provider interests and broader ideologies. A spurious cloak of inevitability serves to promote and justify political choices.


Effect of fresh-frozen plasma resuscitation on cardiopulmonary function and serum protein flux.
The adverse effects of albumin resuscitation on coagulation activity, cardiopulmonary function, and extravascular flux of nonalbumin protein have made fresh-frozen plasma (FFP) an attractive alternate volume expander for hemorrhagic shock. This study addresses the effects of FFP on cardiopulmonary hemodynamics and protein flux. Whole blood was collected three and six weeks before shock, separated into red blood cells (PRBCs) and FFP, and stored. In 24 conditioned splenectomized dogs, resuscitation from reservoir shock of two hours' duration consisted of autologous PRBCs and balanced electrolyte solution (BES) in control dogs and PRBCs, BES, and FFP in plasma-treated dogs. Hemorrhagic shock reduced serum albumin and IgG levels in both groups. Resuscitation with FFP led to a higher cardiac output, left ventricular stroke work (LVSW), and pulmonary capillary wedge pressure (PCWP). The PCWP/LVSW ratio was comparable for both groups. Postshock day 2 serum albumin and IgG levels and lymphatic albumin and IgG concentrations were increased in plasma dogs. Therefore, FFP supplement to PRBC and BES resuscitation does not derange the PCWP/LVSW ratio or reduce intravascular nonalbumin proteins.


Locomotor function of forelimb protractor and retractor muscles of dogs: evidence of strut-like behavior at the shoulder.
The limbs of running mammals are thought to function as inverted struts. When mammals run at constant speed, the ground reaction force vector appears to be directed near the point of rotation of the limb on the body such that there is little or no moment at the joint. If this is true, little or no external work is done at the proximal joints during constant-speed running. This possibility has important implications to the energetics of running and to the coupling of lung ventilation to the locomotor cycle. To test if the forelimb functions as an inverted strut at the shoulder during constant-speed running and to characterize the locomotor function of extrinsic muscles of the forelimb, we monitored changes in the recruitment of six muscles that span the shoulder (the m. pectoralis superficialis descendens, m. pectoralis profundus, m. latissimus dorsi, m. omotransversarius, m. cleidobrachialis and m. trapezius) to controlled manipulations of locomotor forces and moments in trotting dogs (Canis lupus familiaris Linnaeus 1753). Muscle activity was monitored while the dogs trotted at moderate speed (approximately 2 m s(-1)) on a motorized treadmill. Locomotor forces were modified by (1) adding mass to the trunk, (2) inclining the treadmill so that the dogs ran up- and downhill (3) adding mass to the wrists or (4) applying horizontally directed force to the trunk through a leash. When the dogs trotted at constant speed on a level treadmill, the primary protractor muscles of the forelimb exhibited activity during the last part of the ipsilateral support phase and the beginning of swing phase, a pattern that is consistent with the initiation of swing phase but not with active protraction of the limb during the beginning of support phase. Results of the force manipulations were also consistent with the protractor muscles initiating swing phase and contributing to active braking via production of a protractor moment on the forelimb when the dogs decelerate. A similar situation appears to be true for the major retractor muscles of the forelimb. The m. pectoralis profundus and the m. latissimus dorsi were completely silent during the support phase of the ipsilateral limb when the dogs ran unencumbered and exhibited little or no increase in activity when the dogs carried added mass on their backs to increase any retraction torque during the support phase of constant-speed running. The most likely explanation for these observations is that the ground force reaction vector is oriented very close to the fulcrum of the forelimb such that the forelimb functions as a compliant strut at the shoulder when dogs trot at constant speed on level surfaces. Because the moments at the fulcrum of the pectoral girdle appear to be small during the support phase of a trotting step, a case can be made that it is the activity of the extrinsic appendicular muscles that produce the swing phase of the forelimb that explain the coupled phase relationship between ventilatory airflow and the locomotor cycle in trotting dogs.


Neurotransmission in the carotid body: transmitters and modulators between glomus cells and petrosal ganglion nerve terminals.
The carotid body (CB) is the main arterial chemoreceptor. The most accepted model of arterial chemoreception postulates that carotid body glomus (type I) cells are the primary receptors, which are synaptically connected to the nerve terminals of petrosal ganglion (PG) neurons. In response to natural stimuli, glomus cells are expected to release one (or more) transmitter(s) which, acting on the peripheral nerve terminals of processes from chemosensory petrosal neurons, increases the sensory discharge. Among several molecules present in glomus cells, acetylcholine and adenosine nucleotides and dopamine are considered as excitatory transmitter candidates. In this review, we will examine recent evidence supporting the notion that acetylcholine and adenosine 5'-triphosphate are the main excitatory transmitters in the cat and rat carotid bodies. On the other hand, dopamine may act as a modulator of the chemoreception process in the cat, but as an excitatory transmitter in the rabbit carotid body.


[In vitro anti-tumor effect of methotrexate modified by peptide].
This study is to investigate the anti-tumor effect in vitro of methotrexate modified by LH-RH peptide (LH-RH-MTX). LH-RH receptors highly expressing MCF-7 human breast carcinoma cell line and lowly expressing K562 human erythroleukemia cell line were served as the tested cells. The cell proliferation inhibition rates of LH-RH-MTX were detected by MTT colorimetric assay. The effects of LH-RH-MTX on the cell cycle and apoptosis rates were detected by flow cytometry. The inhibition rate of LH-RH-MTX on MCF-7 cells was much higher than that on K562 cells, and the inhibition rate of LH-RH-MTX on MCF-7 cells was much higher than that of free MTX at the same concentration. The inhibition rate of LH-RH-MTX on rat bone marrow mononuclear cells was less than that of free MTX. The number of MCF-7 cells in S phase increased after administration of LH-RH-MTX. The apoptosis rate of LH-RH-MTX group significantly increased compared with that of the control group and MTX group. The relative expression of LHRHR mRNA of LH-RH-MTX group markedly decreased compared with that of the control group and MTX group. LH-RH-MTX is realizable to reduce drug side effects, increase the therapeutic index and achieve tumor-targeted therapy.


A non-major histocompatibility locus determines tissue specificity in the pathogenic process underlying synovial proliferation in a mouse arthropathy model.
The incidence and characteristics of spontaneous ankylosis in the ankle of specific F(1) mice descended from two Fas-deficient strains were reported. Here the coincidence of synovial proliferation and ankylosis in the descendent F(2) mice is reported. To clarify whether the two distinct manifestations are genetically different. An arthropathic group of mice (MCF(2)) were bred by intercrossing MRL/Mp.Fas(lpr)-sap(-)/sap(-) and C3H/He.Fas(lpr) mice. All mice were killed by bleeding under anaesthesia when they were 6 months old. Pathological grades for synovial proliferation were determined by microscopical examination. To obtain a linkage locus, the whole genome of male MCF(2) mice was scanned by using 73 microsatellite markers. Synovial proliferation was equally observed in male and female MCF(2) mice. No correlation was observed between the grades of synovial proliferation and the ankylosis occurring in the MCF(2) mice. A suggestive susceptibility locus was shown in the middle of chromosome 11. This locus was an MRL allele with a recessive inheritance mode. The pathogenic mechanisms of synovial proliferation and ankylosis are genetically different. The present locus is overlapped with some loci associated with rheumatoid arthritis and with others associated with experimental arthritides.


Spectroscopic and mechanistic investigations of dehaloperoxidase B from Amphitrite ornata.
Dehaloperoxidase (DHP) from the terebellid polychaete Amphitrite ornata is a bifunctional enzyme that possesses both hemoglobin and peroxidase activities. Of the two DHP isoenzymes identified to date, much of the recent focus has been on DHP A, whereas very little is known pertaining to the activity, substrate specificity, mechanism of function, or spectroscopic properties of DHP B. Herein, we report the recombinant expression and purification of DHP B, as well as the details of our investigations into its catalytic cycle using biochemical assays, stopped-flow UV-visible, resonance Raman, and rapid freeze-quench electron paramagnetic resonance spectroscopies, and spectroelectrochemistry. Our experimental design reveals mechanistic insights and kinetic descriptions of the dehaloperoxidase mechanism which have not been previously reported for isoenzyme A. Namely, we demonstrate a novel reaction pathway in which the products of the oxidative dehalogenation of trihalophenols (dihaloquinones) are themselves capable of inducing formation of oxyferrous DHP B, and an updated catalytic cycle for DHP is proposed. We further demonstrate that, unlike the traditional monofunctional peroxidases, the oxyferrous state in DHP is a peroxidase-competent starting species, which suggests that the ferric oxidation state may not be an obligatory starting point for the enzyme. The data presented herein provide a link between the peroxidase and oxygen transport activities which furthers our understanding of how this bifunctional enzyme is able to unite its two inherent functions in one system.


Evaluation of Collision Cross Section Calibrants for Structural Analysis of Lipids by Traveling Wave Ion Mobility-Mass Spectrometry.
Collision cross section (CCS) measurement of lipids using traveling wave ion mobility-mass spectrometry (TWIM-MS) is of high interest to the lipidomics field. However, currently available calibrants for CCS measurement using TWIM are predominantly peptides that display quite different physical properties and gas-phase conformations from lipids, which could lead to large CCS calibration errors for lipids. Here we report the direct CCS measurement of a series of phosphatidylcholines (PCs) and phosphatidylethanolamines (PEs) in nitrogen using a drift tube ion mobility (DTIM) instrument and an evaluation of the accuracy and reproducibility of PCs and PEs as CCS calibrants for phospholipids against different classes of calibrants, including polyalanine (PolyAla), tetraalkylammonium salts (TAA), and hexakis(fluoroalkoxy)phosphazines (HFAP), in both positive and negative modes in TWIM-MS analysis. We demonstrate that structurally mismatched calibrants lead to larger errors in calibrated CCS values while the structurally matched calibrants, PCs and PEs, gave highly accurate and reproducible CCS values at different traveling wave parameters. Using the lipid calibrants, the majority of the CCS values of several classes of phospholipids measured by TWIM are within 2% error of the CCS values measured by DTIM. The development of phospholipid CCS calibrants will enable high-accuracy structural studies of lipids and add an additional level of validation in the assignment of identifications in untargeted lipidomics experiments.


Mediator subunits and histone methyltransferase Set2 contribute to Ino2-dependent transcriptional activation of phospholipid biosynthesis in the yeast Saccharomyces cerevisiae.
To activate eukaryotic genes, several pathways which modify chromatin and recruit general factors of the transcriptional machinery are utilized. We investigated the factors required for activation of yeast phospholipid biosynthetic genes, depending on activator protein Ino2 which binds to the inositol/choline-responsive element (ICRE) upstream promoter motif together with its partner protein Ino4. We used a set of 15 strains each defective for one of the non essential subunits of yeast mediator complex and identified med2, med3, med15, med18 and med19 as impaired for inositol biosynthesis. In these mutants, ICRE-dependent gene activation was reduced to 13-22% of the wild-type level. We also demonstrate synthetic growth and activation defects among mediator mutants and mutants lacking defined histone modifications (snf1, gcn5) and transcriptional coactivators (sub1). Analysis of mutants defective for histone methylation (set1, set2 and dot1) and demethylation (jhd1, jhd2, gis1, rph1 and ecm5) revealed the importance of the H3 Lys36-specific Set2 methyltransferase for ICRE-dependent gene expression. Although defined mediator subunits are critical for gene activation, we could not detect their interaction with Ino2. In contrast, Ino2 directly binds to the Set2 histone methyltransferase. Mapping of interaction domains revealed the importance of the SET core domain which was necessary and sufficient for binding Ino2.


Carriage of the zoonotic organism Streptococcus suis in chicken flocks in Vietnam.
Streptococcus suis infections are an emerging zoonotic agent causing severe disease in humans and a major pig pathogen worldwide. We investigated the colonization of S. suis in healthy chickens in different flocks (n = 59) as well as in-contact pigs in farms with S. suis-positive chickens (n = 44) in the Mekong Delta of Vietnam. Streptococcus suis was isolated from 20 (33.9%) chicken flocks and from all pigs investigated. Chicken isolates formed a distinct genotypic cluster compared with pig and human strains, although two chicken isolates (10%) clustered with pig isolates. Chicken isolates had unusually high levels of resistance against tetracycline (100%), clindamycin (100%) and erythromycin (95%); and intermediate resistance against penicillin (35%) and ceftriaxone (15%). Our findings suggest that chickens may potentially represent a source of S. suis infection to in-contact humans and pigs.


Expression of the adenovirus E1A oncogene during cell transformation is sufficient to induce susceptibility to lysis by host inflammatory cells.
Mammalian cells transformed by nononcogenic human adenoviruses exhibit high susceptibility to destruction by host mononuclear inflammatory cells. We have analyzed the viral gene regulation of the susceptibility of transformed cells to lysis by natural killer cells and activated macrophages. Comparisons of target cell lines transformed by overlapping segments of the adenovirus E1-transforming gene region revealed that isolated expression of a single oncogene, E1A, was sufficient to cause increased cytolytic susceptibility in the absence of detectable transformed cell-surface expression of viral transplantation antigens and irrespective of histocompatibility antigen identity between killer cells and target cells. These results suggest that oncogene functions that are not linked to the expression of previously recognized cell-surface target structures may actively induce neoplastic cell elimination by components of the host immune surveillance system.


Vitamin D supplementation in children and young adults with persistent proteinuria secondary to glomerular disease.
Vitamin D deficiency is common in glomerular disease. Supplementation may be ineffective due to ongoing urinary losses of vitamin D binding protein. We sought to determine if daily cholecalciferol supplementation would increase vitamin D concentrations in children with glomerular disease and persistent proteinuria, without adverse effects. Eighteen participants at least 5 years of age with primary glomerular disease and urine protein:creatinine ratio ≥ 0.5 were enrolled from four pediatric nephrology practices to receive cholecalciferol supplementation: 4,000 IU or 2,000 IU per day for serum 25 hydroxyvitamin vitamin D (25OHD) concentrations < 20 ng/mL and 20 ng/mL to < 30 ng/mL, respectively. Measures of vitamin D and mineral metabolism were obtained at baseline and weeks 6 and 12. Multivariable generalized estimating equation (GEE) regression estimated mean percent changes in serum 25OHD concentration. Median baseline 25OHD was 12.8 ng/mL (IQR 9.3, 18.9) and increased to 27.8 ng/mL (20.5, 36.0) at week 6 (p < 0.001) without further significant increase at week 12. A total of 31% of participants had a level ≥ 30 ng/mL at week 12. Supplementation was stopped in two participants at week 6 for mildly elevated calcium and phosphorus, respectively, with subsequent declines in 25OHD of > 20 ng/mL. In the adjusted GEE model, 25OHD was 102% (95% CI: 64, 141) and 96% (95% CI: 51, 140) higher versus baseline at weeks 6 and 12, respectively (p < 0.001). Cholecalciferol supplementation in vitamin D deficient children with glomerular disease and persistent proteinuria safely increases 25OHD concentration. Ideal dosing to fully replete 25OHD concentrations in this population remains unknown. NCT01835639. A higher resolution version of the Graphical abstract is available as Supplementary information.


Upregulation of the growth arrest-specific-2 in recurrent colorectal cancers, and its susceptibility to chemotherapy in a model cell system.
Colorectal cancer (CRC) is one of the most common life-threatening malignances worldwide. CRC relapse markedly decreases the 5-year survival of patients following surgery. Aberrant expression of genes involved in pathways regulating the cell cycle, cell proliferation, or cell death are frequently reported in CRC tumorigenesis. We hypothesized that genes involved in CRC relapse might serve as prognostic indicators. We first evaluated the significance of gene sequences in the feces of patients with CRC relapse by consulting a public database. Tumorigenesis of target tissues was tested through tumor cell growth, cell cycle regulation, and chemotherapeutic efficacy. We found a highly significant correlation between CRC relapse and growth arrest-specific 2 (GAS2) gene expression. Based on cell models, the overexpressed GAS2 was associated with cellular growth rate, cell cycle regulation, and with chemotherapeutic sensitivity. Cell division was impaired by treating cells with 2-[4-(7-chloro-2-quinoxalinyloxy)phenoxy]-propionic acid (XK469), even when the cells were overexpressing GAS2. Thus, downregulation of GAS2 expression might control CRC relapse after curative resection. GAS2 could serve as a noninvasive marker from the feces of patients with prediagnosed CRC. Our findings suggest that GAS2 could have potential clinical applications for predicting early CRC relapse after radical resection, and that XK469 might impair tumor cell division by reducing GAS2 expression or blocking its cellular translocation. This will help in selecting the best therapeutic option, 5-fluorouracil in combination with XK469, for patients overexpressing GAS2 in CRC cells. Thus, GAS2 might act as a prognostic biomolecule and potential therapeutic target in patients with CRC relapse.


RNA cytidine acetyltransferase of small-subunit ribosomal RNA: identification of acetylation sites and the responsible acetyltransferase in fission yeast, Schizosaccharomyces pombe.
The eukaryotic small-subunit (SSU) ribosomal RNA (rRNA) has two evolutionarily conserved acetylcytidines. However, the acetylation sites and the acetyltransferase responsible for the acetylation have not been identified. We performed a comprehensive MS-based analysis covering the entire sequence of the fission yeast, Schizosaccharomyces pombe, SSU rRNA and identified two acetylcytidines at positions 1297 and 1815 in the 3' half of the rRNA. To identify the enzyme responsible for the cytidine acetylation, we searched for an S. pombe gene homologous to TmcA, a bacterial tRNA N-acetyltransferase, and found one potential candidate, Nat10. A temperature-sensitive strain of Nat10 with a mutation in the Walker A type ATP-binding motif abolished the cytidine acetylation in SSU rRNA, and the wild-type Nat10 supplemented to this strain recovered the acetylation, providing evidence that Nat10 is necessary for acetylation of SSU rRNA. The Nat10 mutant strain showed a slow-growth phenotype and was defective in forming the SSU rRNA from the precursor RNA, suggesting that cytidine acetylation is necessary for ribosome assembly.


dUTPase as a platform for antimalarial drug design: structural basis for the selectivity of a class of nucleoside inhibitors.
Pyrimidine metabolism is a major route for therapeutic intervention against malaria. Here we report inhibition and structural studies on the deoxyuridine nucleotidohydrolase from the malaria parasite Plasmodium falciparum (PfdUTPase). We have identified a series of triphenylmethane derivatives of deoxyuridine with antimalarial activity in vitro which inhibit specifically the Plasmodium dUTPase versus the human enzyme. A 2.4 Angstrom crystal structure of PfdUTPase in complex with one of these inhibitors reveals an atypical trimeric enzyme in which the triphenylmethane derivative can be seen to select for PfdUTPase by way of interactions between the trityl group and the side chains of residues Phe46 and Ile117. Immunofluorescence microscopy studies of parasitized red blood cells reveal that enzyme concentrations are highest during the trophozoite/schizont stages, suggesting that PfdUTPase has a major role in DNA replication. Taken together the data show that PfdUTPase may be considered as an antimalarial drug target.


High school dietary dairy intake and teenage acne.
Previous studies suggest possible associations between Western diet and acne. We examined data from the Nurses Health Study II to retrospectively evaluate whether intakes of dairy foods during high school were associated with physician-diagnosed severe teenage acne. We studied 47,355 women who completed questionnaires on high school diet in 1998 and physician-diagnosed severe teenage acne in 1989. We estimated the prevalence ratios and 95% confidence intervals of acne history across categories of intakes. After accounting for age, age at menarche, body mass index, and energy intake, the multivariate prevalence ratio (95% confidence intervals; P value for test of trend) of acne, comparing extreme categories of intake, were: 1.22 (1.03, 1.44; .002) for total milk; 1.12 (1.00, 1.25; .56) for whole milk; 1.16 (1.01, 1.34; .25) for low-fat milk; and 1.44 (1.21, 1.72; .003) for skim milk. Instant breakfast drink, sherbet, cottage cheese, and cream cheese were also positively associated with acne. We found a positive association with acne for intake of total milk and skim milk. We hypothesize that the association with milk may be because of the presence of hormones and bioactive molecules in milk.


Povidone-iodine Solutions Inhibit Cell Migration and Survival of Osteoblasts, Fibroblasts, and Myoblasts.
In vitro laboratory study. The purpose of this study was to identify the effect of dilute povidone-iodine (PVI) solutions on human osteoblast, fibroblast and myoblast cells in vitro. Dilute PVI wound lavage has been used successfully in spine and joint arthroplasty procedures to prevent postoperative surgical site infection, but their biologic effect on host cells is largely unknown. Human primary osteoblasts, fibroblasts, and myoblasts were expanded in cell culture and subjected to various concentrations of PVI (0%, 0.001%, 0.01%, 0.1%, 0.35%, 1%) for 3 minutes. To assess the effect of PVI on cell migration, a scratch assay was performed, in which a "scratch" was made by a standard pipette tip in a cell monolayer following PVI exposure, and time to closure of the scratch was evaluated. Cell survival and proliferation was measured 48 hours post-PVI exposure using a cell viability and cytotoxicity assay. Closure of the scratch defect in all cell monolayers was achieved in <24 hours in untreated controls and following exposure to PVI concentrations <0.1%. The scratch defect remained open indefinitely following exposure to PVI concentrations of ≥0.1%. PVI concentrations <0.1% did not have significant effect on survival rates compared with control for all cell types. Cells exposed to PVI ≥ 0.1% had cell survival rates of less than 6% (P < 0.05). Clinically used concentration of PVI (0.35%) exerts a pronounced cytotoxic effect on osteoblasts, fibroblast, and myoblasts in vitro. Further investigation is required to systematically study the effect of PVI on tissue healing in vivo and also determine a safe and clinically potent concentration for PVI lavage. N/A.


The menopause transition in women living with HIV: current evidence and future avenues of research.
As the life expectancy of people living with HIV improves as a result of antiretroviral therapy, increasing numbers of women living with HIV (WLHIV) are now reaching menopausal age. The menopause transition in WLHIV remains a relatively overlooked area in clinical HIV research. Whilst there is some evidence to suggest that WLHIV experience menopause at an earlier age and that they have more menopausal symptoms, there is no clear consensus in the literature around an impact of HIV infection on either timing or symptomatology of the menopause. Data are also conflicting on whether HIV-related factors such as HIV viral load and CD4 cell count have an impact on the menopause. Furthermore, menopausal symptoms in WLHIV are known to go under-recognised by both healthcare providers and women themselves. There is likely to be a burden of unmet health needs among WLHIV transitioning through the menopause, with significant gaps in the evidence base for their care. With this in mind, we have developed the PRIME study (Positive Transitions Through the Menopause). This mixed-methods observational study will explore, for the first time in the UK, the impact of the menopause on the health and wellbeing of 1500 ethnically diverse WLHIV. In establishing a cohort of women in their midlife and following them up longitudinally, we hope to develop a nuanced understanding of the gendered aspects of ageing and HIV, informing the provision of appropriate services for WLHIV to ensure that they are supported in maintaining optimal health and wellbeing as they get older.


Alzheimer's Disease Assessment Scale--cognitive in clinical practice.
The goal of this article is (a) to review a series of measurement standards for group and individual-patient applications of cognitive tests for Alzheimer's disease (AD); and (b) to review the literature and to use specific data from the NORMACODEM (a project of standardization of cognitive and functional toosl for AD) to evaluate the measurement standards of the Alzheimer's Disease Assessment Scale-Cognitive (ADAS-Cog). Seven measurement standards for group and individual patient use were evaluated: practicality, extent of measurement, depth of measurement, cross-sectional precision, longitudinal precision, validity, and standardization. The results indicated that ADAS-Cog meets practicality standards, that the scoring method is not difficult, but training is needed to administer the test. In comparison with other rating scales, the ADAS-Cog is regarded as more comprehensive, although it is not a substitute for an extensive neuropsychological test. The ADAS-Cog is not a tool that is suitable for primary care physicians, although it is a valuable screening test that has utility in both early detection (diagnosis) and staging of AD, and is useful in the severity range of Global Dementia stages 1 to 6. Interrater reliability coefficients for the ADAS-Cog satisfy recommended standards, and the test shows good internal consistency and reproducibility. Age and education are associated with the ADAS-Cog score. There is, however, a lack of standardization, which restricts the comparability of study results, and test materials remain unpublished.