Emerging Environmental Justice Issues in Nuclear Power and Radioactive Contamination.
Nuclear hazards, linked to both U.S. weapons programs and civilian nuclear power, pose substantial environment justice issues. Nuclear power plant (NPP) reactors produce low-level ionizing radiation, high level nuclear waste, and are subject to catastrophic contamination events. Justice concerns include plant locations and the large potentially exposed populations, as well as issues in siting, nuclear safety, and barriers to public participation. Other justice issues relate to extensive contamination in the U.S. nuclear weapons complex, and the mining and processing industries that have supported it. To approach the topic, first we discuss distributional justice issues of NPP sites in the U.S. and related procedural injustices in siting, operation, and emergency preparedness. Then we discuss justice concerns involving the U.S. nuclear weapons complex and the ways that uranium mining, processing, and weapons development have affected those living downwind, including a substantial American Indian population. Next we examine the problem of high-level nuclear waste and the risk implications of the lack of secure long-term storage. The handling and deposition of toxic nuclear wastes pose new transgenerational justice issues of unprecedented duration, in comparison to any other industry. Finally, we discuss the persistent risks of nuclear technologies and renewable energy alternatives.


Identification and qualification of 500 nuclear, single-copy, orthologous genes for the Eupulmonata (Gastropoda) using transcriptome sequencing and exon capture.
The qualification of orthology is a significant challenge when developing large, multiloci phylogenetic data sets from assembled transcripts. Transcriptome assemblies have various attributes, such as fragmentation, frameshifts and mis-indexing, which pose problems to automated methods of orthology assessment. Here, we identify a set of orthologous single-copy genes from transcriptome assemblies for the land snails and slugs (Eupulmonata) using a thorough approach to orthology determination involving manual alignment curation, gene tree assessment and sequencing from genomic DNA. We qualified the orthology of 500 nuclear, protein-coding genes from the transcriptome assemblies of 21 eupulmonate species to produce the most complete phylogenetic data matrix for a major molluscan lineage to date, both in terms of taxon and character completeness. Exon capture targeting 490 of the 500 genes (those with at least one exon >120 bp) from 22 species of Australian Camaenidae successfully captured sequences of 2825 exons (representing all targeted genes), with only a 3.7% reduction in the data matrix due to the presence of putative paralogs or pseudogenes. The automated pipeline Agalma retrieved the majority of the manually qualified 500 single-copy gene set and identified a further 375 putative single-copy genes, although it failed to account for fragmented transcripts resulting in lower data matrix completeness when considering the original 500 genes. This could potentially explain the minor inconsistencies we observed in the supported topologies for the 21 eupulmonate species between the manually curated and 'Agalma-equivalent' data set (sharing 458 genes). Overall, our study confirms the utility of the 500 gene set to resolve phylogenetic relationships at a range of evolutionary depths and highlights the importance of addressing fragmentation at the homolog alignment stage for probe design.


Effect of recombinant human bone morphogenetic protein-4 dose on bone formation in a rat calvarial defect model.
Bone morphogenetic proteins (BMPs) are being evaluated for periodontal and bone regenerative therapy. The objective of this study was to evaluate the effect of recombinant human bone morphogenetic protein-4 (rhBMP-4) dose on local bone formation in a rat calvaria defect model. Calvarial, 8 mm diameter, critical-size osteotomy defects were created in 140 male Sprague-Dawley rats. Seven groups of 20 animals each received either 1) rhBMP-4 (2.5 microg) in an absorbable collagen sponge (ACS) carrier, 2) rhBMP-4 (5 microg)/ACS, 3) rhBMP-4 (2.5 microg) in a beta-tricalcium phosphate (beta-TCP) carrier, 4) rhBMP-4 (5 microg)/beta-TCP, 5) ACS or 6) beta-TCP carrier controls, or 7) a sham-surgery control, and were evaluated by histologic and histometric parameters following a 2- or 8-week healing interval (10 animals/group/healing interval). Surgical implantation of rhBMP-4/ACS and rhBMP-4/beta-TCP resulted in enhanced local bone formation at both 2 and 8 weeks. Within the dose range examined, rhBMP-4 did not exhibit an appreciable dose-dependent response. Defect closure was not significantly different between the rhBMP-4/ACS and rhBMP-4/beta-TCP groups. New bone area of the rhBMP-4/ beta-TCP group was significantly greater than that of the rhBMP-4/ ACS group; however, bone density in the rhBMP-4/ACS group was significantly greater than that in the rhBMP-4/beta-TCP group at 8 weeks (P < 0.05). rhBMP-4 combined with ACS or beta-TCP has a significant potential to induce bone formation in the rat calvaria defect model. Within the selected rhBMP-4 dose range and observation interval, there appeared to be no meaningful differences in bone formation.


Towards action research with trans women sex workers: Policy, space and social challenges.
Trans women face multiple social, economic and health inequalities and the impact of gender oppression and violence is even more profoundly experienced by trans women sex workers, although in culturally specific ways. This paper presents a pilot study conducted to explore and engage with the context of trans women sex workers in Lido Tre Archi, Italy. In line with the community psychology values of social justice, social change and participation, we outline our engagement process and key challenges observed when attempting action research with such a highly marginalised group. Data were collected in the form of ethnographic notes from informal consultations with different stakeholders and participant observations, and of documents (newspaper articles, picture captions) and were analysed using thematic analysis. Findings are organised in three themes relating to the social, policy and physical context in Lido Tre Archi demonstrating evidence of the contextual challenges and how they intertwine to generate a spiral of marginalisation and social exclusion for the participants. We take a collaborative and reflexive stance in our work and conclude with recommended steps and potential limitations to initiate an action research project.


Tropospheric aerosol optical properties derived from lidar, sun photometer, and optical particle counter measurements.
Tropospheric aerosols have been observed for the period from November 1990 to April 1992 with a lidar, a sun photometer, and an optical particle counter. Variations of aerosol optical thickness derived from the lidar and the sun photometer data and measurements are presented. The simultaneous measurements of these instruments also allowed us to estimate the extinction-to-backscatter ratio (S(1)), which ranged from 20 to 70. Comparison of optical thicknesses derived from both instruments clearly shows the effect of Mt. Pinatubo's eruption and the temporal variation of optical thickness in the stratosphere over 12 km. The possible range of the complex refractive index for the columnar mean aerosols can be deduced from the probable range of S(1) derived by the use of an S(1) diagram as a function of complex refractive index (m). The imaginary part of m can be estimated provided that the real part of m is known.


Acceptability of oral rapid HIV testing at dental clinics in communities with high HIV prevalence in South Florida.
Expanding HIV screening for populations at risk necessitates testing in nontraditional settings. We assessed HIV testing in dental clinics in South Florida, an urban area with the highest rates of HIV diagnoses in the United States in 2015. We explored patients' acceptance of oral HIV rapid tests administered by dental providers and identified reasons for accepting or declining HIV testing. During 2014 and 2015, dentists and hygienists at two federally qualified health center (FQHC) dental clinics who serve racial/ethnic minority patient populations in South Florida were trained to administer oral HIV rapid tests as a part of a routine dental visit. Patients presenting for dental services were offered a rapid HIV test and brief survey regarding their demographics, HIV testing history and behaviors. We enrolled 600 patients (median age = 43 years; IQR: 29-56 years), 45% non-Hispanic black and 35% Hispanic/Latino, 83% graduated high school, and 50% unemployed. Most (85%) accepted oral HIV rapid testing (none tested HIV-positive); 14% had never been tested for HIV. The most common reasons for testing were a desire to know HIV status (56%) and free testing (54%). Among 93 (15%) patients who declined testing, 58% were tested recently and 31% felt confident that they were HIV-negative; however, 74 (80%) who declined testing said they would feel comfortable discussing HIV prevention with their dentist. Additionally, 290 of 600 patients (48%) reported condomless vaginal or anal sex in the past 6 months. Further, among 119 patients who had condomless sex with an HIV-positive partner and/or one whose HIV status was unknown, 98 (82%) accepted the oral HIV test. Dental clinics may provide expanded opportunities for oral HIV rapid testing and conversations about HIV prevention in high HIV prevalence communities.


In your phase: neural phase synchronisation underlies visual imagery of faces.
Mental imagery is the process through which we retrieve and recombine information from our memory to elicit the subjective impression of "seeing with the mind's eye". In the social domain, we imagine other individuals while recalling our encounters with them or modelling alternative social interactions in future. Many studies using imaging and neurophysiological techniques have shown several similarities in brain activity between visual imagery and visual perception, and have identified frontoparietal, occipital and temporal neural components of visual imagery. However, the neural connectivity between these regions during visual imagery of socially relevant stimuli has not been studied. Here we used electroencephalography to investigate neural connectivity and its dynamics between frontal, parietal, occipital and temporal electrodes during visual imagery of faces. We found that voluntary visual imagery of faces is associated with long-range phase synchronisation in the gamma frequency range between frontoparietal electrode pairs and between occipitoparietal electrode pairs. In contrast, no effect of imagery was observed in the connectivity between occipitotemporal electrode pairs. Gamma range synchronisation between occipitoparietal electrode pairs predicted subjective ratings of the contour definition of imagined faces. Furthermore, we found that visual imagery of faces is associated with an increase of short-range frontal synchronisation in the theta frequency range, which temporally preceded the long-range increase in the gamma synchronisation. We speculate that the local frontal synchrony in the theta frequency range might be associated with an effortful top-down mnemonic reactivation of faces. In contrast, the long-range connectivity in the gamma frequency range along the fronto-parieto-occipital axis might be related to the endogenous binding and subjective clarity of facial visual features.


Adipocyte fatty acid-binding protein: regulation of gene expression in vivo and in vitro by an insulin-sensitizing agent.
Pioglitazone, a thiazolidinedione, is a novel antidiabetic compound that can lower blood glucose in diabetic rodents by increasing insulin sensitivity in target tissues. We have previously demonstrated that pioglitazone can enhance the insulin- or insulin-like growth factor-1-regulated differentiation of 3T3-L1 cells, a cell line that undergoes morphological and biochemical differentiation to mature adipocytes [Mol. Pharmacol. 41:393-398 (1992)]. In this study, we have examined the effect of pioglitazone on the expression of the adipocyte fatty acid-binding protein (aFABP) in ob/ob mice and 3T3-L1 cells. Administration of the drug to mice was observed to cause a dose-dependent increase in aFABP mRNA expression in epididymal fat, which was correlated with a decrease in blood glucose and insulin levels. Treatment of 3T3-L1 cells with pioglitazone enhanced aFABP expression in a time-dependent fashion. To explore a possible direct effect of pioglitazone on aFABP expression, a chimeric gene was constructed containing the aFABP promoter fused upstream of the bacterial reporter gene for chloramphenicol acetyltransferase. After transfection into 3T3-L1 cells and selection of stable transformants, regulation of the chimeric gene was studied. Pioglitazone, in combination with insulin or insulin-like growth factor-1, was observed to elicit a dose-dependent increase in expression, indicating a role for pioglitazone in regulating transcription of the aFABP gene. Several thiazolidinedione analogs were tested for their ability to induce the expression of the chimeric gene, and it was found that activity in this assay paralleled the structure-activity relationships observed for enhancement of 3T3-L1 cell differentiation. These observations on control of aFABP gene expression by pioglitazone suggest possible mechanisms by which cellular sensitivity to insulin may be regulated.


Does a high dietary acid content cause bone loss, and can bone loss be prevented with an alkaline diet?
A popular concept in nutrition and lay literature is that of the role of a diet high in acid or protein in the pathogenesis of osteoporosis. A diet rich in fruit and vegetable intake is thought to enhance bone health as the result of its greater potassium and lower "acidic" content than a diet rich in animal protein and sodium. Consequently, there have been a number of studies of diet manipulation to enhance potassium and "alkaline" content of the diet to improve bone density or other parameters of bone health. Although acid loading or an acidic diet featuring a high protein intake may be associated with an increase in calciuria, the evidence supporting a role of these variables in the development of osteoporosis is not consistent. Similarly, intervention studies with a more alkaline diet or use of supplements of potassium citrate or bicarbonate have not consistently shown a bone health benefit. In the elderly, inadequate protein intake is a greater problem for bone health than protein excess.


Effect of dietary palm oils on mammary carcinogenesis in female rats induced by 7,12-dimethylbenz(a)anthracene.
Female Sprague-Dawley rats, 50 days of age, were treated with a single dose of 5 mg of 7,12-dimethylbenz(a)anthracene intragastrically. 3 days after carcinogen treatment, the rats were put on semisynthetic diets containing 20% by weight of corn oil (CO), soybean oil (SBO), crude palm oil (CPO), refined, bleached, deodorized palm oil (RBD PO) and metabisulfite-treated palm oil (MCPO) for 5 months. During the course of experiments, rats fed on different dietary fats had similar rate of growth. Rats fed 20% CO or SBO diet have higher tumor incidence than rats fed on palm oil (PO) diets; however differences of mean tumor latency periods among the groups were not statistically significant. At autopsy, rats fed on high CO or SBO diets had significantly more tumors than rats fed on the three PO diets. Our results showed that high PO diets did not promote chemically induced mammary tumorigenesis in female rats when compared to high CO or SBO diets. CO and SBO differ greatly from the palm oils in their contents of tocopherols, tocotrienols, and carotenes. But further experiments would be required to determine whether the observed differences in tumor incidence and tumor numbers were due to the differences in these minor components or due to the unique triglyceride structure of the palm oils. Analysis of the fatty acid profiles of plasma total lipids of tumor-bearing rats and of the tumor total lipids showed that, with the exception of arachidonic acid, the fatty acid profiles reflect the nature of the dietary fats. At autopsy, there were no differences in the plasma total cholesterol contents among rats fed on different dietary fats, but rats fed on palm oil diets had a significantly higher plasma triglyceride level than that of rats fed CO or SBO diets. As for the tumor lipids, there were no significant differences in the triglyceride, diglyceride, and phospholipid levels when the CO or SBO groups were compared to the palm oil groups.


Loading of an Mcm protein onto DNA replication origins is regulated by Cdc6p and CDKs.
In eukaryotic cells, firing of DNA replication origins normally does not recur until after M phase. This characteristic is thought to be due to the properties of "initiation" proteins like Orc, Cdc6, and Mcms. Using formaldehyde cross-linking, we show that Cdc6p and Mcm7p associate specifically with replication origins during G1 but not during G2 in S. cerevisiae. Mcm7p's association with origins depends on Cdc6p. Ectopic expression of Cdc6p enables it to associate with origins during G2, but this fails to recruit Mcm7p. Our data suggest that the loading of Mcm proteins onto origins is regulated by two mechanisms: first, by Cdc6p occupancy, and second, by S- and M-CDKs, whose activity during S, G2, and M phases prevents Mcm loading.


Isolation and nature of intracellular peptides from a cephalosporin C-producing Cephalosporium sp.
1. Three peptides containing alpha-aminoadipic acid and cysteine have been obtained in small amounts from the mycelium of a Cephalosporium sp. 2. The peptides were precipitated as cuprous mercaptides together with glutathione and resolved from the latter and from each other by preparative paper electrophoresis and chromatography either in the sulphonic acid form or as S-sulphonyl derivatives. From the S-sulphonyl derivatives they were obtained in the thiol form. 3. One peptide (P3) was shown by amino acid analysis and the mass spectrum of the NS-ethoxycarbonyl derivative of its methyl ester to be delta-(l-alpha-aminoadipyl)-l-cysteinyl-d-valine. A second peptide (P2) contained alpha-aminoadipic acid, cysteine, valine and glycine, and the third peptide (P1) contained alpha-aminoadipic acid, cysteine, beta-hydroxyvaline and glycine.


Effects of massage on delayed-onset muscle soreness, swelling, and recovery of muscle function.
Delayed-onset muscle soreness (DOMS) describes muscle pain and tenderness that typically develop several hours postexercise and consist of predominantly eccentric muscle actions, especially if the exercise is unfamiliar. Although DOMS is likely a symptom of eccentric-exercise-induced muscle damage, it does not necessarily reflect muscle damage. Some prophylactic or therapeutic modalities may be effective only for alleviating DOMS, whereas others may enhance recovery of muscle function without affecting DOMS. To test the hypothesis that massage applied after eccentric exercise would effectively alleviate DOMS without affecting muscle function. We used an arm-to-arm comparison model with 2 independent variables (control and massage) and 6 dependent variables (maximal isometric and isokinetic voluntary strength, range of motion, upper arm circumference, plasma creatine kinase activity, and muscle soreness). A 2-way repeated-measures analysis of variance and paired t tests were used to examine differences in changes of the dependent variable over time (before, immediately and 30 minutes after exercise, and 1, 2, 3, 4, 7, 10, and 14 days postexercise) between control and massage conditions. University laboratory. Ten healthy subjects (5 men and 5 women) with no history of upper arm injury and no experience in resistance training. Subjects performed 10 sets of 6 maximal isokinetic (90 degrees x s(-1)) eccentric actions of the elbow flexors with each arm on a dynamometer, separated by 2 weeks. One arm received 10 minutes of massage 3 hours after eccentric exercise; the contralateral arm received no treatment. Maximal voluntary isometric and isokinetic elbow flexor strength, range of motion, upper arm circumference, plasma creatine kinase activity, and muscle soreness. Delayed-onset muscle soreness was significantly less for the massage condition for peak soreness in extending the elbow joint and palpating the brachioradialis muscle (P < .05). Soreness while flexing the elbow joint (P = .07) and palpating the brachialis muscle (P = .06) was also less with massage. Massage treatment had significant effects on plasma creatine kinase activity, with a significantly lower peak value at 4 days postexercise (P < .05), and upper arm circumference, with a significantly smaller increase than the control at 3 and 4 days postexercise (P < .05). However, no significant effects of massage on recovery of muscle strength and ROM were evident. Massage was effective in alleviating DOMS by approximately 30% and reducing swelling, but it had no effects on muscle function.


D-Dimer Levels and Traditional Risk Factors Are Associated With Incident Hypertension Among HIV-Infected Individuals Initiating Antiretroviral Therapy in Uganda.
We sought to describe blood pressure (BP) changes after antiretroviral therapy (ART) initiation and evaluate the association of markers of inflammation with incident hypertension in a cohort of HIV-infected individuals in Uganda. We used mixed effects linear regression to model changes in systolic BP over time among a cohort of HIV-infected individuals initiating ART in Uganda. After exclusion of participants with preexisting hypertension, we identified participants with normal BP throughout follow-up (controls) and those with elevated BP on ≥3 consecutive visits (cases). Before ART initiation, participants had testing for interleukin 6, kynurenine/tryptophan ratio, lipopolysaccharide, soluble CD14, soluble CD163, and D-dimer and those with viral suppression at 6 months during ART had repeat tests. We fit logistic regression models to estimate associations between biomarkers and risk of incident hypertension. In the entire cohort, systolic BP increased by 9.6 mm Hg/yr (95% CI: 7.3 to 11.8) in the first 6 months of ART, then plateaued. Traditional factors: male gender (adjusted odds ratio (AOR) 2.76, 95% CI: 1.34 to 5.68), age (AOR 1.09, 95% CI: 1.04 to 1.13), overweight (AOR 4.48, 95% CI: 1.83 to 10.97), and a CD4 count <100 cells (AOR 3.08, 95% CI: 1.07 to 8.89) were associated with incident hypertension. After adjusting for these, D-dimer levels at month 6 were inversely associated with incident hypertension (AOR 0.61, 95% CI: 0.37 to 0.99). Although not significant, similar associations were seen with sCD14 and kynurenine/tryptophan ratio. BP increases early after ART initiation in Ugandans. Traditional risk factors, rather than immune activation, were associated with incident hypertension in this population.


An exploratory study of the association of acute posttraumatic stress, depression, and pain to cognitive functioning in mild traumatic brain injury.
Few studies have examined whether psychological distress and pain affect cognitive functioning in the acute to subacute phase (up to 30 days postinjury) following mild traumatic brain injury (mTBI). The current study explored whether acute posttraumatic stress, depression, and pain were associated with performance on a task of selective and sustained attention completed under conditions of increasing cognitive demands (standard, auditory distraction, and dual-task), and on tests of working memory, memory, processing speed, reaction time (RT), and verbal fluency. At a mean of 2.87 days (SD = 2.32) postinjury, 50 adult mTBI participants, consecutive admissions to a Level 1 trauma hospital, completed neuropsychological tests and self-report measures of acute posttraumatic stress, depression, and pain. A series of canonical correlation analyses was used to explore the relationships of a common set of psychological variables to various sets of neuropsychological variables. Significant results were found on the task of selective and sustained attention. Strong relationships were found between psychological variables and speed (r(c) = .56, p = .02) and psychological variables and accuracy (r(c) = .68, p = .002). Pain and acute posttraumatic stress were associated with higher speed scores (reflecting more correctly marked targets) under standard conditions. Acute posttraumatic stress was associated with lower accuracy scores across all task conditions. Moderate but nonsignificant associations were found between psychological variables and most cognitive tasks. Acute posttraumatic stress and pain show strong associations with selective and sustained attention following mTBI.


Direct access to midlevel dental providers: an evidence synthesis.
Direct access is a term that describes the ability of patients to seek health care from midlevel dental providers (MLDPs) without first seeing a dentist. The objective of this study was to synthesize the evidence for the effects and costs of direct access to MLDPs in a primary dental care environment and assess the attitudes of various stakeholders to this method of care delivery. The literature was examined for descriptive, observational, and experimental study designs to examine the evidence for direct access in dentistry. Electronic searches were undertaken of the Database of Abstracts of Reviews of Effectiveness, bibliographic subscription databases, open access databases, and the gray literature. The search identified 371 records, although the extent of experimental evidence was limited. The majority of included studies were descriptive and recorded the subjective views of different stakeholders, following the introduction of the policy. The limited extent of experimental evidence regarding direct access to MLDPs contrasts with their widespread use across Europe, the United States, and the southern hemisphere. Suggestions are made for a research program to improve the evidence base for direct access.


Vitamin D3 and deconvoluting a rash.
BACKGROUNDAdverse drug reactions are unpredictable immunologic events presenting frequent challenges to clinical management. Systemically administered cholecalciferol (vitamin D3) has immunomodulatory properties. In this randomized, double-blinded, placebo-controlled interventional trial of healthy human adults, we investigated the clinical and molecular immunomodulatory effects of a single high dose of oral vitamin D3 on an experimentally induced chemical rash.METHODSSkin inflammation was induced with topical nitrogen mustard (NM) in 28 participants. Participant-specific inflammatory responses to NM alone were characterized using clinical measures, serum studies, and skin tissue analysis over the next week. All participants underwent repeat NM exposure to the opposite arm and then received placebo or 200,000 IU cholecalciferol intervention. The complete rash reaction was followed by multi-omic analysis, clinical measures, and serum studies over 6 weeks.RESULTSCholecalciferol mitigated acute inflammation in all participants and achieved 6 weeks of durable responses. Integrative analysis of skin and blood identified an unexpected divergence in response severity to NM, corroborated by systemic neutrophilia and significant histopathologic and clinical differences. Multi-omic and pathway analyses revealed a 3-biomarker signature (CCL20, CCL2, CXCL8) unique to exaggerated responders that is suppressed by cholecalciferol and implicates IL-17 signaling involvement.CONCLUSIONHigh-dose systemic cholecalciferol may be an effective treatment for severe reactions to topical chemotherapy. Our findings have broad implications for cholecalciferol as an antiinflammatory intervention against the development of exaggerated immune responses.TRIAL REGISTRATIONclinicaltrials.gov (NCT02968446).FUNDINGNIH and National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS; grants U01AR064144, U01AR071168, P30 AR075049, U54 AR079795, and P30 AR039750 (CWRU)).


Histones cause aggregation and fusion of lipid vesicles containing phosphatidylinositol-4-phosphate.
In a previous article, we demonstrated that histones (H1 or histone octamers) interact with negatively charged bilayers and induce extensive aggregation of vesicles containing phosphatidylinositol-4-phosphate (PIP) and, to a lesser extent, vesicles containing phosphatidylinositol (PI). Here, we found that vesicles containing PIP, but not those containing PI, can undergo fusion induced by histones. Fusion was demonstrated through the observation of intervesicular mixing of total lipids and inner monolayer lipids, and by ultrastructural and confocal microscopy studies. Moreover, in both PI- and PIP-containing vesicles, histones caused permeabilization and release of vesicular aqueous contents, but the leakage mechanism was different (all-or-none for PI and graded release for PIP vesicles). These results indicate that histones could play a role in the remodeling of the nuclear envelope that takes place during the mitotic cycle.


Calcium binding capacity of erythrocyte membrane in human hypertension.
The cell membrane calcium binding capacity of genetically hypertensive rats is reduced when measured in the presence of the submicromolar calcium concentrations proper of intracellular environment. The present work, performed as an ancillary study to an epidemiological survey on an entire population, aimed to investigate the existence of a similar abnormality in human hypertension. Calcium binding to the erythrocyte membrane was measured in clinically healthy normotensive (n = 12) and hypertensive individuals (n = 24). For this purpose, a filtration technique was used, based on the determination of 45Ca bound to the erythrocyte membrane in the presence of free calcium concentrations (40 nmol/l and 1 mumol/l), which are similar to those of the intracellular environment. The intra-assay technical error was determined on 35 duplicate samples and, when expressed as percent of the mean, was 24.1 at the 40 nmol/l concentration and 16.8 at the 1 mumol/l concentration. Membranes of untreated hypertensive patients, at both calcium concentrations, bound significantly less calcium than the control group. Treated and untreated hypertensive individuals had comparable values of membrane calcium binding capacity. Membranes of the treated hypertensive group bound less calcium than those of the normotensive group at both calcium concentrations, but the difference was statistically significant only in the presence of 40 nmol/l free calcium. A significant positive correlation was observed between the calcium binding capacity at 40 nmol/l concentration and that at 1 mumol/l in the treated and untreated hypertensive groups (r = 0.73 and 0.75, respectively; 0.51 for the normotensive group). These findings support the hypothesis that a cell membrane abnormality is detectable in some hypertensive patients.(ABSTRACT TRUNCATED AT 250 WORDS)


Cost-Effectiveness of Percutaneous Closure of the Left Atrial Appendage in Atrial Fibrillation Based on Results From PROTECT AF Versus PREVAIL.
Randomized trials of left atrial appendage (LAA) closure with the Watchman device have shown varying results, and its cost effectiveness compared with anticoagulation has not been evaluated using all available contemporary trial data. We used a Markov decision model to estimate lifetime quality-adjusted survival, costs, and cost effectiveness of LAA closure with Watchman, compared directly with warfarin and indirectly with dabigatran, using data from the long-term (mean 3.8 year) follow-up of Percutaneous Closure of the Left Atrial Appendage Versus Warfarin Therapy for Prevention of Stroke in Patients With Atrial Fibrillation (PROTECT AF) and Prospective Randomized Evaluation of the Watchman LAA Closure Device in Patients With Atrial Fibrillation (PREVAIL) randomized trials. Using data from PROTECT AF, the incremental cost-effectiveness ratios compared with warfarin and dabigatran were $20 486 and $23 422 per quality-adjusted life year, respectively. Using data from PREVAIL, LAA closure was dominated by warfarin and dabigatran, meaning that it was less effective (8.44, 8.54, and 8.59 quality-adjusted life years, respectively) and more costly. At a willingness-to-pay threshold of $50 000 per quality-adjusted life year, LAA closure was cost effective 90% and 9% of the time under PROTECT AF and PREVAIL assumptions, respectively. These results were sensitive to the rates of ischemic stroke and intracranial hemorrhage for LAA closure and medical anticoagulation. Using data from the PROTECT AF trial, LAA closure with the Watchman device was cost effective; using PREVAIL trial data, Watchman was more costly and less effective than warfarin and dabigatran. PROTECT AF enrolled more patients and has substantially longer follow-up time, allowing greater statistical certainty with the cost-effectiveness results. However, longer-term trial results and postmarketing surveillance of major adverse events will be vital to determining the value of the Watchman in clinical practice.


Structural characterization of small molecular ions by ion mobility mass spectrometry in nitrogen drift gas: improving the accuracy of trajectory method calculations.
The investigation of ion structures based on a combination of ion mobility mass spectrometry (IM-MS) experiments and theoretical collision cross section (CCS) calculations has become important to many fields of research. However, the accuracy of current CCS calculations for ions in nitrogen drift gas limits the information content of many experiments. In particular, few studies have evaluated and attempted to improve the theoretical tools for CCS calculation in nitrogen drift gas. In this study, based on high-quality experimental measurements and theoretical modeling, a comprehensive evaluation of various aspects of CCS calculations in nitrogen drift gas is performed. It is shown that the modification of the ion-nitrogen van der Waals (vdW) interaction potential enables accurate CCS predictions of 29 small ions with ca. 3% maximum relative error. The present method exhibits no apparent systematic bias with respect to ion CCS (size) and dipole moment, suggesting that the method adequately describes the long-range interactions between the ions and the buffer gas. However, the method shows limitations in reproducing experimental CCS at low temperatures (<150 K) and for macromolecular ions, and calculations for these cases should be complemented by CCS calculation methods in helium drift gas. This study presents an accurate and well-characterized CCS calculation method for ions in nitrogen drift gas that is expected to become an important tool for ion structural characterization and molecular identification. The experimental values reported here also provide a foundation for future studies aiming at developing more efficient computational tools.


Development of fetal yawn compared with non-yawn mouth openings from 24-36 weeks gestation.
Although some research suggests that fetuses yawn, others disagree arguing that is it simple mouth opening. Furthermore there is no developmental account of fetal yawning compared with simple mouth opening. The aim of the present study was to establish in a repeated measures design the development of fetal yawning compared with simple mouth opening. Video recordings were made of the fetal face and upper torso visualized by means of 4D full frontal or facial profile ultrasound recordings. Fifteen healthy fetuses were scanned four times at 24, 28, 32 and 36 weeks gestation. Yawning was distinguished from non-yawning in terms of the length of time it took to reach the apex of the mouth stretch, with yawns being defined as more than 50% of the total time observed. To assess changes in frequency, a Poisson mixed effects model was fitted to the count of number of yawn and simple mouth opening events with age and gender as fixed effects, and person as a random effect. For both yawns and simple mouth openings a smooth varying age effect was significant. The number of yawns observed declined with age from 28 weeks gestation, whereas simple mouth openings were less frequent and the decline was observed from 24 weeks. Gender was not significant either for yawn and simple mouth openings. Yawning can be reliably distinguished from other forms of mouth opening with the potential of using yawning as an index of fetal healthy development.


Recent advances in the treatment of amyotrophic lateral sclerosis. Emphasis on kynurenine pathway inhibitors.
Amyotrophic lateral sclerosis (ALS) is an adult onset, progressive and fatal motor neuron degenerative disease [1]. The aetiology of ALS is currently unknown, though strongly suggested to be multifactorial. Recently, the kynurenine pathway (KP) has emerged as a potential contributing factor [2]. The KP is a major route for the metabolism of tryptophan, generating neuroactive intermediates in the process. These catabolites include the excitotoxic N-methyl-D-aspartate (NMDA) receptor agonist, quinolinic acid (QUIN) [3] and the neuroprotective NMDA receptor antagonist, kynurenic acid (KYNA) [4,5]. These catabolites appear to play a key role in the communication between the nervous and immune systems, and also in modulating cell proliferation and tissue function [6]. As the cause of ALS is still unknown, there is presently no efficient treatment for it. Currently, Riluzole is the drug of choice but its effect is relatively modest [7]. Targeting the KP, hence, could offer a new therapeutic option to improve ALS treatment [8]. Several drugs that block the KP are already under investigation by our laboratory and others, some of which are in or about to enter clinical trials for other diseases. For example, the KP inhibitors, Teriflunomide (Sanofi-Aventis) and Laquinimod (Teva Neuroscience). Recently, a KP inhibitor has also reached the Japan market as an immunomodulative drug [9]: Tranilast/Rizaben (Angiogen Ltd.) is an anthranilic acid derivative [8]. Finally, the 8-hydroxyquinolinine metal attenuating compounds, Clioquinol and PBT2, interestingly have close structural similarity with KYNA and QUIN. Such drugs would open a new and important therapeutic door for ALS.


Home fortification in emergency response and transition programming: experiences in Aceh and Nias, Indonesia.
Improving the nutritional status of an emergency-affected population is essential, because undernutrition increases infection risk and is linked to more than 50% of childhood deaths in developing countries. Emergency food aid addresses nutritional needs, including micronutrient deficiencies, but is provided only for a limited time and uses few items, so the needs of specific target groups are often not fully met. To describe the post-tsunami experience with distribution of Vitalita Sprinkles in Aceh and Nias and to analyze the monitoring data gathered for the emergency response. International agencies such as Helen Keller International and partners provided micronutrients in response to the tsunami emergency in Aceh and Nias and to analyze the monitoring data gathered for the emergency response. In March and April 2006, the percentage of children aged 6 months to 59 months who had consumed Vitalita in the previous month was more than 70% in 5 of 11 districts evaluated, 40% to 70% in another five districts, and 32% in one district. An independent survey found 25% less anemia among recipients. Almost all mothers interviewed during March to April 2006 (96.3%) had heard about Vitalita and recognized its packaging, 69% said that Vitalita contained vitamins for under-fives, 86% knew the appropriate target group for Vitalita, and 83% said it had to be mixed with solid food. Of the 26% that reported not having given Vitalita to their child in the last month, 90.5% said that their child did not like it. CONCLUSIONS AND LESSONS LEARNED: Because the product, including its concept, was new, appropriate information and training was required as well as a thorough introduction to the beneficiaries, particularly on proper use, to ensure acceptance. This experience demonstrates that providing micronutrients as part of emergency relief and transition programming, as recently recommended by the World Health Organization/UNICEF/World Food Program, is feasible.


Corticosteroids for the common cold.
The common cold is a frequent illness, which, although benign and self-limiting, results in many consultations to primary care and considerable loss of school or work days. Current symptomatic treatments have limited benefit. Corticosteroids are an effective treatment in other upper respiratory tract infections and their anti-inflammatory effects may also be beneficial in the common cold. To compare corticosteroids versus usual care for the common cold on clinical response rates in children and adults. We searched the Cochrane Central Register of Controlled Trials (CENTRAL) 2012, Issue 5 which includes the Acute Respiratory Infections (ARI) Group's Specialised Register, the Database of Reviews of Effects (DARE) 2012, Issue 4 and the NHS Health Economics Database 2012, Issue 5; MEDLINE (1948 to May week 2, 2012) and EMBASE (January 2010 to May 2012). Randomised, double-blind, controlled trials comparing corticosteroids to placebo or to standard clinical management. Two review authors independently extracted data and assessed trial quality. We were unable to perform meta-analysis and instead analysed results using narrative description of the available evidence. We included two trials (253 participants). Both compared intranasal corticosteroids to placebo; no trials studied oral corticosteroids. No benefit of intranasal corticosteroids was demonstrated for duration or severity of symptoms. In one trial of 54 participants, the number of symptomatic days was 10.3 in the placebo group, compared to 10.7 in those using intranasal corticosteroids (P = 0.72). A second trial of 199 participants reported no significant differences in duration of symptoms. There were no differences reported in terms of: adverse events; complications (one case of sinusitis, one case of acute otitis media, both in corticosteroid groups); presence of rhinovirus in nasal aspirates; or treatment for secondary infections. Neither trial reported our primary outcome measure of percentage of participants with resolution at different time points. A lack of comparable outcome measures meant we were unable to combine the data. Current evidence does not support the use of intranasal corticosteroids for symptomatic relief from the common cold. However, there were only two trials and limited statistical power. Further large randomised placebo-controlled trials in adults and children are required to answer this question.


[Artificial nutrition in hyperglycemia and diabetes mellitus in critically ill patients].
The need to strictly control glucose levels, even in nondiabetic patients, has recently emerged following the publication of the results that indicate the possibility of reducing the morbidity and mortality in critically ill patients. Since hyperglycemia is one of the most frequent metabolic impairments in these patients, insulin therapy is a necessity in most of the cases. In order to prevent hyperglycemia and its associated complications, nutritional support must be adjusted to the patient's requirements, avoiding hyponutrition. Whenever possible, nutrients supply should be done through the digestive route. Parenteral nutrition is more often accompanied by hyperglycemia and requires an increase in insulin dosage to control it. There are two types of enteral diets designed to help controlling hyperglycemic conditions: carbohydrates rich diets, and fat rich diets. In general terms, carbohydrates rich diets may be recommended in type 1 diabetic patients who are in a stable condition, and fat rich diets in type 2 diabetes and in stress hyperglycemia. In both cases, the use of low glycemic index carbohydrates is recommended. Protein intake should be adjusted to the patients' metabolic stress level. In diabetic patients with acute disease, an increase in antioxidants intake is recommended.


The experiences of mothers of children with autism spectrum disorder: Managing family routines and mothers' health and wellbeing.
Families of children with autism spectrum disorder (ASD) use family routines to provide predictability and structure to support occupational engagement of their family members. Mothers assume the major role to orchestrate occupations in constructing family routines, which may impact their health and wellbeing. However, the experiences of mothers in managing family routines and their health and wellbeing have not been the main focus in previous research. Thus, this study explored the experiences of mothers of children with ASD in managing family routines and their perceptions of the impact of family routines on their health and wellbeing. An interpretive phenomenological approach was used. Twenty mothers of children with ASD, aged between 28 and 56 years, participated in semi-structured interviews. Data were transcribed verbatim and each transcript was analysed. Five themes that summarise mothers' perceptions towards health and wellbeing when managing family routines emerged: (i) Keeping on track keeping healthy; (ii) My life is busy, because I do everything for everyone else; (iii) Keeping on track all the time is tiring or frustrating; (iv) Looking after my family by looking after myself; and (v) I am not perfect and it is OK. This study highlighted the substantial efforts required in constructing family routines that may be at the cost of mothers' health and wellbeing. However, mothers may be able to cope with everyday demands in managing family routines by changing their perspectives. By integrating 'me-time' activities in family routines, mothers may be able to support their own health and wellbeing. Mothers' values and needs are reflected in family routines; hence, thorough understanding of family routines may be a key to support mothers' occupational engagement.


The Untapped Potential of Patient and Family Engagement in the Organization of Critical Care.
There is growing interest in patient and family participation in critical care-not just as part of the bedside, but as part of educational and management organization and infrastructure. This offers tremendous opportunities for change but carries risk to patients, families, and the institution. The objective is to provide a concise definitive review of patient and family organizational participation in critical care as a high-risk population and other vulnerable groups. A pragmatic, codesigned model for critical care is offered as a suggested approach for clinicians, researchers, and policy-makers. To inform this review, a systematic search of Ovid Medline, PubMed, and Embase was undertaken in April 2016 using the MeSH terms: patient participation and critical care. A second search was undertaken in PubMed using the terms: patient participation and organizational models to search for other examples of engagement in vulnerable populations. We explicitly did not seek to include discussions of bedside patient-family engagement or shared decision-making. Two reviewers screened citations independently. Included studies either actively partnered with patients and families or described a model of engagement in critical care and other vulnerable populations. Data or description of how patient and family engagement occurred and/or description of model were extracted into a standardized form. There was limited evidence of patient and family engagement in critical care although key recommendations can be drawn from included studies. Patient and family engagement is occurring in other vulnerable populations although there are few described models and none which address issues of risk. A model of patient and family engagement in critical care does not exist, and we propose a pragmatic, codesigned model that takes into account issues of psychologic safety in this population. Significant opportunity exists to document processes of engagement that reflect a changing paradigm of healthcare delivery.


Electronic Cigarettes and Youth in the United States: A Call to Action (at the Local, National and Global Levels).
E-cigarettes have emerged and soared in popularity in the past ten years, making them the most common tobacco product used among youth in the United States (US). In this review, we discuss what the Surgeon General has called a public health "epidemic"-the precipitous increase in youth use of e-cigarettes and the health consequences of this behavior. Further, we review tobacco control policy efforts (e.g., Tobacco 21, banning flavors, advertising restrictions, and clean indoor air laws)-efforts proven to be critical in reducing cigarette smoking and smoking-related disease and death among US children and adults-including their potential and challenges regarding managing and mitigating the emergence of e-cigarettes. Finally, we close with a discussion of the efforts of transnational tobacco companies to rebrand themselves using e-cigarettes and other new products.


Mechanistic insights into a classic wonder drug--aspirin.
Aspirin, one of the oldest and most common anti-inflammatory agents, has recently been shown to reduce cancer risks. The principal pharmacological effects of aspirin are known to arise from its covalent modification of cyclooxygenase-2 (COX-2) through acetylation of Ser530, but the detailed mechanism of its biochemical action and specificity remains to be elucidated. In this work, we have filled this gap by employing a state-of-the-art computational approach, Born-Oppenheimer molecular dynamics simulations with ab initio quantum mechanical/molecular mechanical potential and umbrella sampling. Our studies have characterized a substrate-assisted inhibition mechanism for aspirin acetylating COX: it proceeds in two successive stages with a metastable tetrahedral intermediate, in which the carboxyl group of aspirin serves as the general base. The computational results confirmed that aspirin would be 10-100 times more potent against COX-1 than against COX-2, and revealed that this inhibition specificity between the two COX isoforms can be attributed mainly to the difference in kinetics rate of the covalent inhibition reaction, not the aspirin-binding step. The structural origin of this differential inhibition of the COX enzymes by aspirin has also been elucidated.


Recurrent wheeze and its relationship with lung function and airway inflammation in preschool children: a cross-sectional study in South Korea.
Relationship between recurrent wheeze and airway function and inflammation in preschool children is not fully known. To investigate the relationship between recurrent wheeze and airway inflammation, lung function, airway hyper-reactivity (AHR) and atopy in preschool children. Observational study, comparing forced expiratory volume in 1 s (FEV1), forced vital capacity (FVC) and mid-forced expiratory flow (FEF25%-75%), dose-response slope (DRS), exhaled nitric oxide (eNO) and atopic sensitisation between children with recurrent wheeze and those without. Population-based, cross-sectional study in Seoul and the Gyeonggi province of Korea conducted as a government-funded programme to perform standardised measurement of the prevalence of allergic diseases, and related factors, in preschool children. 900 children aged 4-6 years. PRIMARY AND SECONDARY OUTCOME MEASURES: eNO, FEV1/FVC, FEF25%-75%, DRS, atopic sensitisation and allergic diseases. Children completed the modified International Study of Asthma and Allergies in Childhood questionnaire and underwent eNO assessments, spirometry, methacholine bronchial provocation tests and skin prick tests. Recurrent wheeze was defined as having a lifetime wheeze of more than three episodes, based on the questionnaire. The frequency of hospitalisation and emergency room visits was also obtained by means of the questionnaire. 'Current' wheeze was defined as having symptoms or treatments within the past 12 months. The prevalence of recurrent wheeze was 13.4%. Children with recurrent wheeze showed a higher prevalence of lifetime or current allergic rhinitis (p=0.01 and p=0.002, respectively) and lifetime atopic dermatitis (p=0.007). Children with recurrent wheeze showed lower FEV1/FVC (p=0.033) and FEF25%-75% (p=0.004), and higher eNO levels (p=0.013) than those without recurrent wheeze. However, the DRS, prevalence of atopic sensitisation and serum IgE levels were not significantly different between the two groups. Recurrent wheeze in preschool children may be associated with airway inflammation and diminished airway function, but not with AHR or atopy.


The roles of entropy and kinetics in structure prediction.
Here we continue our efforts to use methods developed in the folding mechanism community to both better understand and improve structure prediction. Our previous work demonstrated that Rosetta's coarse-grained potentials may actually impede accurate structure prediction at full-atom resolution. Based on this work we postulated that it may be time to work completely at full-atom resolution but that doing so may require more careful attention to the kinetics of convergence. To explore the possibility of working entirely at full-atom resolution, we apply enhanced sampling algorithms and the free energy theory developed in the folding mechanism community to full-atom protein structure prediction with the prominent Rosetta package. We find that Rosetta's full-atom scoring function is indeed able to recognize diverse protein native states and that there is a strong correlation between score and Calpha RMSD to the native state. However, we also show that there is a huge entropic barrier to folding under this potential and the kinetics of folding are extremely slow. We then exploit this new understanding to suggest ways to improve structure prediction. Based on this work we hypothesize that structure prediction may be improved by taking a more physical approach, i.e. considering the nature of the model thermodynamics and kinetics which result from structure prediction simulations.


Quantification of In Planta Zymoseptoria tritici Progression Through Different Infection Phases and Related Association with Components of Aggressiveness.
In planta growth of Zymoseptoria tritici, causal agent of Septoria tritici blotch of wheat, during the infection process has remained an understudied topic due to the long symptomless latent period before the emergence of fruiting bodies. In this study, we attempted to understand the relationship between in planta growth of Z. tritici relative to the primary components of aggressiveness, i.e., latent period and pycnidia coverage in regard to contrasting host resistance. We tested isolates collected from Ireland against the susceptible cultivar Gallant and cultivar Stigg, which has strong partial resistance. A clear isolate-host interaction effect (F = 3.018; P = 0.005, and F = 6.008; P < 0.001) for latent period and pycnidia coverage, respectively, was identified. Furthermore, during the early infection phase of latency from 5 to 11 days postinoculation (dpi), in planta growth rate of fungal biomass was significantly (F = 30.06; P < 0.001) more affected by host resistance than isolate specificity (F = 1.27; P = 0.27), indicating the importance of host resistance in the early infection phase. In planta Z. tritici growth rates in cultivar Gallant spiked between 11 and 16 dpi followed by a continuous fall onward, whereas in cultivar Stigg it was slowly progressive in nature. From correlation and regression analysis, we found that the in planta growth rate preceding the average latent period of cultivar Gallant has more influence on latency duration and pycnidia production. Likewise, correlation between component of aggressiveness and in planta growth rate of pathogen supports our understanding of aggressiveness to be driven by the pathogen's multiplication capacity within host tissue.


Vagal Nerve Stimulation: Indications, Implantation, and Outcomes.
Vagal nerve stimulation (VNS) therapy is a surgical treatment that involves the implantation of a device to electrically stimulate the vagus nerve. It is indicated as an adjunctive treatment of epilepsy that is refractory to antiepileptic medications and for treatment-resistant depression. The exact mechanism by which VNS achieves its effects is not known, but various mechanisms have been proposed, including afferent vagal projections to seizure-generating regions of the brain and desynchronization of hypersynchronized cortical activity. The most common complications of VNS therapy include hoarseness, throat pain/dysphagia, coughing, and shortness of breath.


Lewis, secretor, and ABO phenotypes, and sulfomucin expression in gastric intestinal metaplasia.
The closely interrelated Lewis, secretor, and ABO phenotypes have long been linked to the risk of peptic ulcers and gastric cancer and may modulate the interaction between Helicobacter pylori and the gastric surface epithelium. We explored the association between the expression of sulfomucins in gastric intestinal metaplasia, a known marker of preneoplastic progression, and the expression of Lewis, secretor, and ABO phenotypes, in 523 subjects from Nariño, Colombia, and 856 subjects from northern Spain. In both study populations, Lewis (a+/b-) and nonsecretor phenotypes showed a significant positive association with the expression of sulfomucins (odds ratios, 2.4 and 2.6, respectively).


Topologically associating domains are stable units of replication-timing regulation.
Eukaryotic chromosomes replicate in a temporal order known as the replication-timing program. In mammals, replication timing is cell-type-specific with at least half the genome switching replication timing during development, primarily in units of 400-800 kilobases ('replication domains'), whose positions are preserved in different cell types, conserved between species, and appear to confine long-range effects of chromosome rearrangements. Early and late replication correlate, respectively, with open and closed three-dimensional chromatin compartments identified by high-resolution chromosome conformation capture (Hi-C), and, to a lesser extent, late replication correlates with lamina-associated domains (LADs). Recent Hi-C mapping has unveiled substructure within chromatin compartments called topologically associating domains (TADs) that are largely conserved in their positions between cell types and are similar in size to replication domains. However, TADs can be further sub-stratified into smaller domains, challenging the significance of structures at any particular scale. Moreover, attempts to reconcile TADs and LADs to replication-timing data have not revealed a common, underlying domain structure. Here we localize boundaries of replication domains to the early-replicating border of replication-timing transitions and map their positions in 18 human and 13 mouse cell types. We demonstrate that, collectively, replication domain boundaries share a near one-to-one correlation with TAD boundaries, whereas within a cell type, adjacent TADs that replicate at similar times obscure replication domain boundaries, largely accounting for the previously reported lack of alignment. Moreover, cell-type-specific replication timing of TADs partitions the genome into two large-scale sub-nuclear compartments revealing that replication-timing transitions are indistinguishable from late-replicating regions in chromatin composition and lamina association and accounting for the reduced correlation of replication timing to LADs and heterochromatin. Our results reconcile cell-type-specific sub-nuclear compartmentalization and replication timing with developmentally stable structural domains and offer a unified model for large-scale chromosome structure and function.


Autofluorescence of normal, benign, and malignant ovarian tissues: a pilot study.
The objective of this study is to evaluate the efficacy of laser-induced fluorescence (LIF) data obtained at 325-nm pulsed laser excitation for the discrimination of normal, benign, and malignant ovarian tissues. Several studies have reported that the autofluorescence technique has a high specificity and sensitivity for discrimination between diseased and non-diseased tissues of various cancers, and also has the advantages of being non-invasive and producing a real-time diagnosis. When using this technique on ovarian tissues in most of the previously reported studies, multivariate statistical tools were used and classification analyses were carried out. Autofluorescence spectra of normal, benign, and malignant ovarian tissues were recorded with 325-nm pulsed laser excitation in the spectral region from 350-600 nm in vitro. The spectral analysis for discrimination between the different types of tissues was carried out using principal component analysis (PCA)-based non-parametric k-nearest neighbor (k-NN) analysis. A total of 97 (34 normal, 33 benign, and 30 malignant) spectra were obtained from 22 subjects with normal, benign, and malignant tissues. The discrimination analysis of data using a PCA-based k-NN algorithm showed very good discrimination. The performance of the analysis was evaluated by calculating statistical parameters, specificity, sensitivity, and accuracy and were found to be 100%, 90.90%, and 94.2%, respectively. The results show that the discrimination of normal, benign, and malignant ovarian conditions can be achieved quite successfully using LIF.


Ursodeoxycholic acid for liver disease associated with cystic fibrosis: a double-blind multicenter trial. The Italian Group for the Study of Ursodeoxycholic Acid in Cystic Fibrosis.
Liver disease is increasingly recognized as a major cause of morbidity in cystic fibrosis (CF). Preliminary data suggest that ursodeoxycholic acid (UDCA) may be beneficial for treatment of this manifestation. We performed a double-blind, multicenter trial in these patients to establish efficacy and safety of UDCA in terms of the improvement of clinical and nutritional indicators besides standard liver function tests. We also intended to establish whether taurine supplementation has a beneficial effect in patients receiving UDCA. From June to December 1990, we enrolled in 12 centers 55 CF patients with liver disease (39 male subjects; median age, 13.8 years). They were randomly assigned to receive for 1 year one of the following treatments: UDCA (15 mg/kg body weight daily) plus taurine (30 mg/kg body weight daily), UDCA plus placebo, placebo plus taurine, or double placebo. Clinical and laboratory evaluations were performed every 3 months. After 1 year, deterioration of overall clinical conditions, as indicated by the Shwachman-Kulczycki score (SKS), occurred in patients who received placebo but not in those who received UDCA (P = .025). Patients treated with UDCA also showed an improvement in gamma-glutamyl transpeptidase (GGT) (P = .004) and 5'-nucleotidase (P = .006) levels. Treatment with taurine was followed by a significant increase in serum prealbumin levels (P = .053), a trend toward a reduction in fat malabsorption, and no effect on the biochemical profile. No severe side effects occurred with any treatment. Thus, we concluded that UDCA administration improves clinical and biochemical parameters in CF patients with liver disease. Taurine supplementation may be indicated in patients with severe pancreatic insufficiency and poor nutritional status.


An investigation into the aspirations and experiences of newly appointed dual diagnosis workers.
This qualitative, exploratory study was designed to explore a sample of eight recently appointed dual diagnosis workers' (DDWs) perceptions of their new role and function in Northern Ireland (NI). A semi-structured interview was used and respondents were assured that their anonymity/rights would be protected. All of the narratives were shown to the respondents for their approval prior to going to press. The transcripts were analysed by using a tried and tested analytical framework. Seven key categories emerged from the findings relating to the DDWs perceptions of their: (1)understanding of the term dual diagnosis; (2) hopes; (3) fears; (4) support in their new role; (5) key clinical issues; (6) the positioning of the service; and (7) their overall role and function. This is a new and important area of work in NI. However, to date, no research has been carried out on the topic in the province. Consequently, the findings from this small study could go some way towards helping to shape the future direction of, and bring about some universality to the provision of the dual diagnosis service within different National Health Service Trusts in NI. Further research is required on this new and growing service as well as on the service users' perceptions of the care provided by DDWs. The study will be followed up on an annual basis for 3 years to provide longitudinal data. Generalization of findings requires caution because of the small sample size.


Towards the use of microfluidics for individual embryo culture.
Mammalian embryo development is still relatively inefficient in vitro. Much research has been conducted on the chemical environment, or culture medium, surrounding the embryo, but little attention has been given to the actual physical culture environment, which has changed very little over the years. The application of microfluidics to embryo production in vitro is a tantalising approach that may alleviate some of the limits that traditional microdrop culture places on embryo development and research into gamete and embryo physiology. These devices may lead to enhanced in vitro embryo development and quality by more closely mimicking the in vivo environment. Initial work in this area is promising and gives us proof-of-principle that these unique microfluidic systems may indeed be applicable to in vitro culture of gametes and embryos. The present paper reviews the advantages of microfluidics for in vitro embryo production: how the platforms are manufactured, the current uses of microfluidics in assisted reproduction, static v. dynamic culture environments, individual gamete and embryo culture and the future directions of microfluidic application to in vitro embryo production and manipulation. Finally, preliminary data from our laboratory using a new microfluidic well insert for porcine, bovine and murine embryo culture is discussed.


Alcohol consumption and survival after a breast cancer diagnosis: a literature-based meta-analysis and collaborative analysis of data for 29,239 cases.
Evidence for an association of alcohol consumption with prognosis after a diagnosis of breast cancer has been inconsistent. We have reviewed and summarized the published evidence and evaluated the association using individual patient data from multiple case cohorts. A MEDLINE search to identify studies published up to January 2013 was performed. We combined published estimates of survival time for "moderate drinkers" versus nondrinkers. An analysis of individual participant data using Cox regression was carried out using data from 11 case cohorts. We identified 11 published studies suitable for inclusion in the meta-analysis. Moderate postdiagnosis alcohol consumption was not associated with overall survival [HR, 0.95; 95% confidence interval (CI), 0.85-1.05], but there was some evidence of better survival associated with prediagnosis consumption (HR, 0.80; 95% CI, 0.73-0.88). Individual data on alcohol consumption for 29,239 cases with 4,839 deaths were available from the 11 case cohorts, all of which had data on estrogen receptor (ER) status. For women with ER-positive disease, there was little evidence that pre- or postdiagnosis alcohol consumption is associated with breast cancer-specific mortality, with some evidence of a negative association with all-cause mortality. On the basis of a single study, moderate postdiagnosis alcohol intake was associated with a small reduction in breast cancer-specific mortality for women with ER-negative disease. There was no association with prediagnosis intake for women with ER-negative disease. There was little evidence that pre- or post-diagnosis alcohol consumption is associated with breast cancer-specific mortality for women with ER-positive disease. There was weak evidence that moderate post-diagnosis alcohol intake is associated with a small reduction in breast cancer-specific mortality in ER-negative disease. Considering the totality of the evidence, moderate postdiagnosis alcohol consumption is unlikely to have a major adverse effect on the survival of women with breast cancer.


Three-dimensional magnetic resonance microscopy of pulmonary solitary tumors in transgenic mice.
We attempted to accurately detect pulmonary solitary tumors and other complicated pulmonary disorders in aging inbred transgenic mice by cardiac- and respiratory-gated MR microscopy at 4.7 Tesla. A comparison of in vivo MR images with histological results demonstrated that submillimeter lung tumors and most of the nontumor lesions could be detected by screening with two-dimensional (2D) gradient echo (GRE) imaging. Subsequently performed 2D spin-echo (SE) imaging provided higher image contrast, which distinguished the tumors from the surrounding complications. On the 3D GRE images and the generated maximum intensity projection (MIP) and volume-rendered (VR) images, proper spatial localization of solitary tumors relative to the orientation of the pulmonary vessels was exhibited, and the tumor volume could also be measured. This promising method is noninvasive and has the potential to eventually replace invasive histopathology because it obviates the need to kill groups of animals at multiple time points.


Identification of the Serine Biosynthesis Pathway as a Critical Component of BRAF Inhibitor Resistance of Melanoma, Pancreatic, and Non-Small Cell Lung Cancer Cells.
Metastatic melanoma cells commonly acquire resistance to BRAF V600E inhibitors (BRAFi). In this study, we identified serine biosynthesis as a critical mechanism of resistance. Proteomic assays revealed differential protein expression of serine biosynthetic enzymes PHGDH, PSPH, and PSAT1 following vemurafenib (BRAFi) treatment in sensitive versus acquired resistant melanoma cells. Ablation of PHGDH via siRNA sensitized acquired resistant cells to vemurafenib. Inhibiting the folate cycle, directly downstream of serine synthesis, with methotrexate also displayed similar sensitization. Using the DNA-damaging drug gemcitabine, we show that gemcitabine pretreatment sensitized resistant melanoma cells to BRAFis vemurafenib and dabrafenib. We extended our findings to BRAF WT tumor cell lines that are intrinsically resistant to vemurafenib and dabrafenib. Pretreatment of pancreatic cancer and non-small cell lung cancer cell lines with sublethal doses of 50 and 5 nmol/L of gemcitabine, respectively, enhanced killing by both vemurafenib and dabrafenib. The novel aspects of this study are the direct identification of serine biosynthesis as a critical mechanism of BRAF V600E inhibitor resistance and the first successful example of using gemcitabine + BRAFis in combination to kill previously drug-resistant cancer cells, creating the translational potential of pretreatment with gemcitabine prior to BRAFi treatment of tumor cells to reverse resistance within the mutational profile and the WT. Mol Cancer Ther; 16(8); 1596-609. ©2017 AACR.


Measuring leg muscle and fat mass in humans: comparison of CT and dual-energy X-ray absorptiometry.
Dual-energy X-ray absorptiometry (DEXA) is reported to be inferior to computed tomography (CT) to measure changes in appendicular soft tissue composition. We compared CT- and DEXA-measured thigh muscle and fat mass to evaluate the random and systematic discrepancies between these two methods. Thigh skeletal muscle area (single-slice CT) was suboptimally (r(2) = 0.74, P < 0.0001) related to DEXA-measured thigh fat-free mass (FFM). In contrast, thigh muscle and adipose tissue volumes (multislice CT) were highly related to DEXA-measured thigh FFM and fat (both r(2) = 0.96, P < 0.0001). DEXA-measured leg fat was significantly less than multislice-CT-measured leg adipose tissue volume, whereas multislice-CT-measured leg muscle mass was less (P < 0.0001) than DEXA-measured leg FFM. The systematic discrepancies between the two approaches were consistent with the 10-15% nonfat components of adipose tissue. In conclusion, CT and DEXA measures of appendicular soft tissue are highly related. Systematic differences between DEXA and CT likely relate to the underlying principles of the techniques.


Indonesian prostate cancer risk calculator (IPCRC): an application for predicting prostate cancer risk (a multicenter study).
to develop a prediction risk model of prostate cancer based on Indonesia population. we included all benign prostate hyperthrophy (BPH) and PCa patients who had prostate biopsy and prostatectomy between January 2009 and December 2013 from 5 urology centers in Indonesia. The relationship between the possibility of PCa with the following variables including: age; PSA level, prostate volume (by transabdominal ultrasound or transrectal ultrasound) and digital rectal examination (DRE) finding. We calculated a predictive scoring equation to predict the possibility of PCa using chi-square analysis, Kolmogorov-Smirnov test, multiple logistic regression and ROC curve. Then, we designed an application for predicting prostate cancer risk called Indonesian Prostate Cancer Risk Calculator (IPCRC). there were 784 PCa and 1173 BPH patients were used for developing the risk calculator in our study. The mean ages, PSA and prostate volume are 66.9±8.1 years old; 72.4±248.9 ng/ml and 49.6±28.2 ml, respectively. Abnormal DRE was found in 637 PCa and 56 BPH. We included age, PSA level, abnormal DRE finding (all showed significant p<0.05 in univariate model). Additionally, although not significant, we included prostate volume (p=0.157) due to its clinical importance. The corrected ROC analysis showed AUC 0.935, sensitivity of 90.1% and specificity 80% in predicting the prostate cancer in our population. we have developed the Indonesian Prostate Cancer Risk Calculator which includes age, PSA, DRE, and prostate volume as its variables. Future prospective study to validate the risk calculator is needed.


Temporal aggregation impacts on epidemiological simulations employing microcontact data.
Microcontact datasets gathered automatically by electronic devices have the potential augment the study of the spread of contagious disease by providing detailed representations of the study population's contact dynamics. However, the impact of data collection experimental design on the subsequent simulation studies has not been adequately addressed. In particular, the impact of study duration and contact dynamics data aggregation on the ultimate outcome of epidemiological models has not been studied in detail, leaving the potential for erroneous conclusions to be made based on simulation outcomes. We employ a previously published data set covering 36 participants for 92 days and a previously published agent-based H1N1 infection model to analyze the impact of contact dynamics representation on the simulated outcome of H1N1 transmission. We compared simulated attack rates resulting from the empirically recorded contact dynamics (ground truth), aggregated, typical day, and artificially generated synthetic networks. No aggregation or sampling policy tested was able to reliably reproduce results from the ground-truth full dynamic network. For the population under study, typical day experimental designs - which extrapolate from data collected over a brief period - exhibited too high a variance to produce consistent results. Aggregated data representations systematically overestimated disease burden, and synthetic networks only reproduced the ground truth case when fitting errors systemically underestimated the total contact, compensating for the systemic overestimation from aggregation. The interdepedendencies of contact dynamics and disease transmission require that detailed contact dynamics data be employed to secure high fidelity in simulation outcomes of disease burden in at least some populations. This finding serves as motivation for larger, longer and more socially diverse contact dynamics tracing experiments and as a caution to researchers employing calibrated aggregate synthetic representations of contact dynamics in simulation, as the calibration may underestimate disease parameters to compensate for the overestimation of disease burden imposed by the aggregate contact network representation.


Resting-state connectivity of the sustained attention network correlates with disease duration in idiopathic generalized epilepsy.
In idiopathic generalized epilepsy (IGE), a normal electroencephalogram between generalized spike and wave (GSW) discharges is believed to reflect normal brain function. However, some studies indicate that even excluding GSW-related errors, IGE patients perform poorly on sustained attention task, the deficit being worse as a function of disease duration. We hypothesized that at least in a subset of structures which are normally involved in sustained attention, resting-state functional connectivity (FC) is different in IGE patients compared to controls and that some of the changes are related to disease duration. Seeds were selected based on a sustained attention study in controls. Resting-state functional magnetic resonance imaging (fMRI) data was obtained from 14 IGE patients and 14 matched controls. After physiological noise removal, the mean time-series of each seed was used as a regressor in a general linear model to detect regions that showed correlation with the seed. In patients, duration factor was defined based on epilepsy duration. Between-group differences weighted by the duration factor were evaluated with mixed-effects model. Correlation was then evaluated in IGE patients between the FC, averaged over each significant cluster, and the duration factor. Eight of 18 seeds showed significant difference in FC across groups. However, only for seeds in the medial superior frontal and precentral gyri and in the medial prefrontal area, average FC taken over significant clusters showed high correlation with the duration factor. These 3 seeds showed changes in FC respectively with the premotor and superior frontal gyrus, the dorsal premotor, and the supplementary motor area plus precentral gyrus. Alterations of FC in IGE patients are not limited to the frontal areas. However, as indicated by specificity analysis, patients with long history of disease show changes in FC mainly within the frontal areas.


Human cloning and embryo research: the 2003 John J. Conley Lecture on medical ethics.
The author, a member of the U.S. President's Council on Bioethics, discusses ethical issues raised by human cloning, whether for purposes of bringing babies to birth or for research purposes. He first argues that every cloned human embryo is a new, distinct, and enduring organism, belonging to the species Homo sapiens, and directing its own development toward maturity. He then distinguishes between two types of capacities belonging to individual organisms belonging to this species, an immediately exerciseable capacity and a basic natural capacity that develops over time. He argues that it is the second type of capacity that is the ground for full moral respect, and that this capacity (and its concomitant degree of respect) belongs to cloned human embryos no less than to adult human beings. He then considers and rejects counter-arguments to his position, including the suggestion that the capacity of embryos is equivalent to the capacity of somatic cells, that full human rights are afforded only to human organisms with functioning brains, that the possibility of twinning diminishes the moral status of embryos, that the fact that people do not typically mourn the loss of early embryos implies that they have a diminished moral status, that the fact that early spontaneous abortions occur frequently diminishes the moral status of embryos, and that his arguments depend upon a concept of ensoulment. He concludes that if the moral status of cloned human embryos is equivalent to that of adults, then public policy should be based upon this assumption.


Tubulin glycylases and glutamylases have distinct functions in stabilization and motility of ependymal cilia.
Microtubules are subject to a variety of posttranslational modifications that potentially regulate cytoskeletal functions. Two modifications, glutamylation and glycylation, are highly enriched in the axonemes of most eukaryotes, and might therefore play particularly important roles in cilia and flagella. Here we systematically analyze the dynamics of glutamylation and glycylation in developing mouse ependymal cilia and the expression of the corresponding enzymes in the brain. By systematically screening enzymes of the TTLL family for specific functions in ependymal cilia, we demonstrate that the glycylating enzymes TTLL3 and TTLL8 were required for stability and maintenance of ependymal cilia, whereas the polyglutamylase TTLL6 was necessary for coordinated beating behavior. Our work provides evidence for a functional separation of glutamylating and glycylating enzymes in mammalian ependymal cilia. It further advances the elucidation of the functions of tubulin posttranslational modifications in motile cilia of the mammalian brain and their potential importance in brain development and disease.


Assessment of Antibodies Induced by Multivalent Transmission-Blocking Malaria Vaccines.
A malaria transmission-blocking vaccine would be a critical tool in achieving malaria elimination and eradication. By using chimpanzee adenovirus serotype 63 and modified vaccinia virus Ankara viral vectored vaccines, we investigated whether incorporating two antigens into one vaccine would result in higher transmission-reducing activity than one antigen. We demonstrated that when Pfs25 was administered with other antigens Pfs28 or Pfs230C, either concurrently as a mixed vaccine or co-expressed as a dual-antigen vaccine, the antibody response in mice to each antigen was comparable to a monoantigen vaccine, without immunological interference. However, we found that the transmission-reducing activity (functional activity) of dual-antigen vaccines was not additive. Dual-antigen vaccines generally only elicited similar transmission-reducing activity to monoantigen vaccines and in one instance had lower transmission-reducing activity. We found that despite the lack of immunological interference of dual-antigen vaccines, they are still not as effective at blocking malaria transmission as Pfs25-IMX313, the current leading candidate for viral vectored vaccines. Pfs25-IMX313 elicited similar quality antibodies to dual-antigen vaccines, but higher antibody titers.


Fecal excretion of a novel human circovirus, TT virus, in healthy children.
The role of TT virus (TTV) as a human pathogen is unclear, as is the mode of TTV transmission. To determine the prevalence of TTV infection and the possible fecal-oral route of transmission, we analyzed fecal specimens from 67 healthy, nontransfused children for TTV DNA sequences by heminested PCR, using the NG and T primer sets. The overall prevalence of TTV fecal excretion was 22.4% (15 of 67), with the T primer set (19.4%) being more sensitive than the NG primer set (10.4%). TTV prevalence based on gender or ethnicity showed no significant differences. None of seven children in the 0- to 6-month age group had detectable TTV in feces. Of three sets of siblings, two unrelated sets of twins, ages 33 and 37 months, were negative for fecal TTV DNA, while the third set of siblings, ages 99 and 35 months, was positive. The absence of TTV in the feces of children younger than 6 months and the high prevalence (40%) in children 7 to 12 months of age is consistent with age-specific acquisition of TTV infection by the nonparenteral route. TTV genotypes 1, 3, 4, and 5 were represented in our study population. TTV-positive siblings had TTV genotypes 1 and 4, suggesting unrelated environmental sources of TTV infection. This observation suggests a possible time frame for TTV acquisition in children which coincides with increased interaction with their environment and increased susceptibility to infectious agents.


NHIN, RHIOs, and Public Health.
Public health plays a critical role in forming the building blocks for community or regional health-information sharing, which is essential to the long-term viability of a Nationwide Health Information Network (NHIN) and the Regional Health Information Organizations (RHIOs). By contributing to its visions, policies, processes, standards, and needs/requirements, public health will close the loop within an NHIN and the RHIOs environment. In this article we illustrate public health's essential role in an NHIN and the RHIOs by examining the mutual benefits to healthcare and public health.


Bone mineral content is positively correlated to n-3 fatty acids in the femur of growing rats.
The present study was conducted to determine whether provision of preformed dietary docosapentaenoic acid (DPAn-6) can replace DHA for normal long bone growth as assessed by dual-energy X-ray absorptiometry for mineral content (BMC). A newly modified artificial rearing method was employed to generate n-3 fatty acid-deficient rats. Except the dam-reared (DR; 3.1 % alpha-linolenic acid) group, newborn pups were separated from their mothers at age 2 d and given artificial rat milk containing linoleic acid (LA), or LA supplemented with 1 % DHA (22 : 6n-3; DHA), 1 % DPAn-6 (DPA), or 1 % DHA plus 0.4 % DPAn-6 (DHA/DPA). The rats were later weaned onto similar pelleted diets. At adulthood, the rats were euthanised and bones (femur, tibia, and lumbar vertebrae) collected for tissue fatty acid analysis and bone mineral density (BMD) determination. The analyses showed that long bones such as femur and tibia in DPAn-6-treated rats contained higher DPAn-6 content and generally had the lowest BMC and BMD values. Hence, DPAn-6 did not replace DHA for normal bone growth and maximal BMC in femur, indicating an indispensible role of DHA in bone health. In conclusion, DHA accumulates in the osteoblast-rich and nerve-abundant periosteum of femur; DHA but not EPA appears to be a vital constituent of marrow and periosteum of healthy modelling bone; and both DHA and total n-3 PUFA strongly correlate to BMC.


Experimental chemonucleolysis with recombinant human matrix metalloproteinase 7 in human herniated discs and dogs.
Chemonucleolysis has been proposed as a less invasive technique than surgery for patients with lumbar disc herniation. Once chymopapain had been approved as a chemonucleolysis drug, it was withdrawn because of serious complications. A novel agent with fewer complications would be desirable. The purpose of this study was to investigate the effects of recombinant human matrix metalloproteinase 7 (rhMMP-7) in experimental chemonucleolysis in vitro and in vivo and examine its effects on tissue damage. The study design is the experimental study using human herniated discs and enzyme substrates in vitro and dogs in vivo. The effects of rhMMP-7 on the degradation of human herniated discs were examined by measuring the wet weight in vitro. The correlations between the decrease in wet weight by rhMMP-7 and the conditions associated with herniated discs were also analyzed. The effects of rhMMP-7 on the proteoglycan and water contents were respectively examined with alcian blue staining and T2-weighted magnetic resonance imaging at 7 days after intradiscal injection in dogs. The distribution of [125I]-labeled rhMMP-7 was investigated by autoradioluminography at 7 days after intradiscal injection in dogs. An epidural injection study with rhMMP-7 was performed to evaluate the effects on the tissue damage around the discs at 1 and 13 weeks after the treatment in dogs. The Type 1 and 2 collagen cleavage rates were measured and compared with those of aggrecan in vitro. Recombinant human matrix metalloproteinase 7 concentration dependently decreased the wet weight of herniated discs in vitro. The decrease in wet weight of the discs by rhMMP-7 did not significantly correlate with the conditions associated with herniated discs. Intradiscal injection of rhMMP-7 reduced the proteoglycan and water contents, with an increase in the serum keratan sulfate levels. Radioactivity of [125I]-labeled rhMMP-7 was detected in the nucleus pulposus and annulus fibrosus but not in the muscle. Epidural injection of rhMMP-7 had no effect on the injection site or the nerve tissues. The Type 1 and 2 collagen cleavage rates of rhMMP-7 were 1,000-fold weaker than those of aggrecan. This study demonstrated experimental chemonucleolysis with rhMMP-7 in vitro and in vivo. The effects of rhMMP-7 were not affected by the conditions associated with herniated discs. The epidural injection study together with the autoradioluminography and in vitro enzyme assay suggests that intradiscal injection of rhMMP-7 may not induce tissue damage around the discs because of its distribution and substrate selectivity. Recombinant human matrix metalloproteinase 7 may be a novel and promising chemonucleolysis agent.


Hunter-gatherers as models in public health.
Hunter-gatherer populations are remarkable for their excellent metabolic and cardiovascular health and thus are often used as models in public health, in an effort to understand the root, evolutionary causes of non-communicable diseases. Here, we review recent work on health, activity, energetics and diet among hunter-gatherers and other small-scale societies (e.g. subsistence farmers, horticulturalists and pastoralists), as well as recent fossil and archaeological discoveries, to provide a more comprehensive perspective on lifestyle and health in these populations. We supplement these analyses with new data from the Hadza, a hunter-gatherer population in northern Tanzania. Longevity among small-scale populations approaches that of industrialized populations, and metabolic and cardiovascular disease are rare. Obesity prevalence is very low (<5%), and mean body fat percentage is modest (women: 24-28%, men: 9-18%). Activity levels are high, exceeding 100 min d-1 of moderate and vigorous physical activity, but daily energy expenditures are similar to industrialized populations. Diets in hunter-gatherer and other small-scale societies tend to be less energy dense and richer in fibre and micronutrients than modern diets but are not invariably low carbohydrate as sometimes argued. A more integrative understanding of hunter-gatherer health and lifestyle, including elements beyond diet and activity, will improve public health efforts in industrialized populations.


Comparison of systemic and local interactions between the arbuscular mycorrhizal fungus Funneliformis mosseae and the root pathogen Aphanomyces euteiches in Medicago truncatula.
It has been shown in a number of pathosystems that arbuscular mycorrhizal (AM) fungi confer resistance against root pathogens, including in interactions between Medicago truncatula and the root rot-causing oomycete Aphanomyces euteiches. For the current study of these interactions, a split root system was established for plant marker gene analysis in order to study systemic defense responses and to compare them with local interactions in conventional pot cultures. It turned out, however, that split root systems and pot cultures were in different physiological stages. Genes for pathogenesis-related proteins and for enzymes involved in flavonoid biosynthesis were generally more highly expressed in split root systems, accompanied by changes in RNA accumulation for genes encoding enzymes involved in phytohormone biosynthesis. Against expectations, the pathogen showed increased activity in these split root systems when the AM fungus Funneliformis mosseae was present separately in the distal part of the roots. Gene expression analysis revealed that this is associated in the pathogen-infected compartment with a systemic down-regulation of a gene coding for isochorismate synthase (ICS), a key enzyme of salicylic acid biosynthesis. At the same time, transcripts of genes encoding pathogenesis-related proteins and for enzymes involved in the biosynthesis of flavonoids accumulated to lower levels. In conventional pot cultures showing decreased A. euteiches activity in the presence of the AM fungus, the ICS gene was down regulated only if both the AM fungus and the pathogen were present in the root system. Such negative priming of salicylic acid biosynthesis could result in increased activities of jasmonate-regulated defense responses and could explain mycorrhiza-induced resistance. Altogether, this study shows that the split root system does not reflect a systemic interaction between F. mosseae and A. euteiches in M. truncatula and indicates the importance of testing such systems prior to the analysis of mycorrhiza-induced resistance.


Overexpression of Glutamate Decarboxylase in Mesenchymal Stem Cells Enhances Their Immunosuppressive Properties and Increases GABA and Nitric Oxide Levels.
The neurotransmitter GABA has been recently identified as a potent immunosuppressive agent that targets both innate and adaptive immune systems and prevents disease progression of several autoimmunity models. Mesenchymal stem cells (MSCs) are self-renewing progenitor cells that differentiate into various cell types under specific conditions, including neurons. In addition, MSC possess strong immunosuppressive capabilities. Upon cytokine priming, undifferentiated MSC suppress T-cell proliferation via cell-to-cell contact mechanisms and the secretion of soluble factors like nitric oxide, prostaglandin E2 and IDO. Although MSC and MSC-derived neuron-like cells express some GABAergic markers in vitro, the role for GABAergic signaling in MSC-mediated immunosuppression remains completely unexplored. Here, we demonstrate that pro-inflammatory cytokines selectively regulate GAD-67 expression in murine bone marrow-MSC. However, expression of GAD-65 is required for maximal GABA release by MSC. Gain of function experiments using GAD-67 and GAD-65 co-expression demonstrates that GAD increases immunosuppressive function in the absence of pro-inflammatory licensing. Moreover, GAD expression in MSC evokes an increase in both GABA and NO levels in the supernatants of co-cultured MSC with activated splenocytes. Notably, the increase in NO levels by GAD expression was not observed in cultures of isolated MSC expressing GAD, suggesting crosstalk between these two pathways in the setting of immunosuppression. These results indicate that GAD expression increases MSC-mediated immunosuppression via secretion of immunosuppressive agents. Our findings may help reconsider GABAergic activation in MSC for immunological disorders.


Moderate hypothermia in the management of resistant automatic tachycardias in children.
Automatic focus tachycardias are often resistant to electrical and pharmacological treatment. Moderate systemic hypothermia (32-34 degrees C) may reduce the tachycardia rate in children with His bundle tachycardia after cardiac surgery. The case notes of seven children with automatic focus tachycardias treated with hypothermia were reviewed. Six had His bundle tachycardia after cardiac surgery and one had ectopic atrial tachycardia; all had signs of low cardiac output. Hypothermia led to a reduction in heart rate in all patients (from 211 (28) (mean (SD] to 146 (5) beats/minute, p less than 0.001), with rises in systolic blood pressure (from 74 (14) mm Hg to 97 (10) mm Hg, p less than 0.01) and hourly urine output (from 0.5 (0.4) ml/kg to 4.6 (2.8) ml/kg, p less than 0.02). No direct adverse effects were noted. The arrhythmia did not resolve in three children, who died (two with His bundle tachycardia after Fontan procedures and one with ectopic atrial tachycardia); the other four regained sinus rhythm which was maintained at follow up of 3-13 (mean 9) months. Moderate systemic hypothermia led to slowing of the arrhythmia rate and an improvement in cardiac output in patients with resistant automatic focus tachycardias. It can be used to improve the haemodynamic condition while other measures of arrhythmia control are being pursued or until spontaneous recovery of normal rhythm.


Mixing of Injectable Fillers: A National Survey.
The mixing of hyaluronic acid or calcium hydroxylapatite fillers with normal saline, plain lidocaine, or lidocaine with epinephrine before injection is a familiar practice among dermatologists. However, the frequency of this practice and rationale behind it has not been well studied. To better elucidate the clinical practice of mixing fillers with other solutions before injecting among dermatologists. A survey was electronically distributed to members of the American Society for Dermatologic Surgery. Four hundred seventy-five dermatologists responded to the survey. Thirty-five percent of respondents mix fillers before injection. Solutions used were as follows: plain lidocaine (44%), lidocaine with epinephrine (36%), normal saline (30%), and sterile water (7%). Respondents mix filler for the following reasons: to decrease viscosity (40%), increase anesthesia (30%), decrease swelling (17%), and increase volume (13%). Despite the lack of evidence, more than one-third of dermatologists surveyed mix fillers with other solutions before injection. Plain lidocaine is most commonly used. The top reason for mixing fillers is to decrease viscosity and facilitate ease of injection. More scientific data are needed to support this practice and better understand the biophysical changes that occur when mixing fillers with other solutions.