Positive network assortativity of influenza vaccination at a high school: implications for outbreak risk and herd immunity.
Schools are known to play a significant role in the spread of influenza. High vaccination coverage can reduce infectious disease spread within schools and the wider community through vaccine-induced immunity in vaccinated individuals and through the indirect effects afforded by herd immunity. In general, herd immunity is greatest when vaccination coverage is highest, but clusters of unvaccinated individuals can reduce herd immunity. Here, we empirically assess the extent of such clustering by measuring whether vaccinated individuals are randomly distributed or demonstrate positive assortativity across a United States high school contact network. Using computational models based on these empirical measurements, we further assess the impact of assortativity on influenza disease dynamics. We found that the contact network was positively assortative with respect to influenza vaccination: unvaccinated individuals tended to be in contact more often with other unvaccinated individuals than with vaccinated individuals, and these effects were most pronounced when we analyzed contact data collected over multiple days. Of note, unvaccinated males contributed substantially more than unvaccinated females towards the measured positive vaccination assortativity. Influenza simulation models using a positively assortative network resulted in larger average outbreak size, and outbreaks were more likely, compared to an otherwise identical network where vaccinated individuals were not clustered. These findings highlight the importance of understanding and addressing heterogeneities in seasonal influenza vaccine uptake for prevention of large, protracted school-based outbreaks of influenza, in addition to continued efforts to increase overall vaccine coverage.


Optimization of alginate purification using polyvinylidene difluoride membrane filtration: Effects on immunogenicity and biocompatibility of three-dimensional alginate scaffolds.
Sodium alginate is an effective biomaterial for tissue engineering applications. Non-purified alginate is contaminated with protein, lipopolysaccharide, DNA, and RNA, which could elicit adverse immunological reactions. We developed a purification protocol to generate biocompatible alginate based on (a) activated charcoal treatment, (b) use of hydrophobic membrane filtration (we used hydrophobic polyvinylidene difluoride membranes to remove organic contaminants), (c) dialysis, and finally (d) ethanol precipitation. Using this approach, we could omit pre-treatment with chloroform and significantly reduce the quantities of reagents used. Purification resulted in reduction of residual protein by 70% down to 0.315 mg/g, DNA by 62% down to 1.28 µg/g, and RNA by 61% down to less than 10 µg/g, respectively. Lipopolysaccharide levels were reduced by >90% to less than 125 EU/g. Purified alginate did not induce splenocyte proliferation in vitro. Three-dimensional scaffolds generated from purified alginate did not elicit a significant foreign body reaction, fibrotic overgrowth, or macrophage infiltration 4 weeks after implantation. This study describes a simplified and economical alginate purification method that results in alginate purity, which meets clinically useful criteria.


Cartilage oligomeric matrix protein neoepitope in the synovial fluid of horses with acute lameness: A new biomarker for the early stages of osteoarthritis.
Clinical tools to diagnose the early changes of osteoarthritis (OA) that occur in the articular cartilage are lacking. We sought to identify and quantify a novel cartilage oligomeric matrix protein (COMP) neoepitope in the synovial fluid from the joints of healthy horses and those with different stages of OA. In vitro quantitative proteomics and assay development with application in synovial fluids samples obtained from biobanks of well-characterised horses. Articular cartilage explants were incubated with or without interleukin-1β for 25 days. Media were analysed via quantitative proteomics. Synovial fluid was obtained from either normal joints (n = 15) or joints causing lameness (n = 17) or with structural OA lesions (n = 7) and analysed for concentrations of the COMP neoepitope using a custom-developed inhibition enzyme-linked immunosorbent assay (ELISA). Explants were immunostained with polyclonal antibodies against COMP and the COMP neoepitopes. Semitryptic COMP peptides were identified and quantified in cell culture media from cartilage explants. A rabbit polyclonal antibody was raised against the neoepitope of the N-terminal portion of one COMP fragment (sequence SGPTHEGVC). An inhibition ELISA was developed to quantify the COMP neoepitope in synovial fluid. The mean concentration of the COMP neoepitope significantly increased in the synovial fluid from the joints responsible for acute lameness compared with normal joints and the joints of chronically lame horses and in joints with chronic structural OA. Immunolabelling for the COMP neoepitope revealed a pericellular staining in the interleukin-1β-stimulated explants. The ELISA is based on polyclonal antisera rather than a monoclonal antibody. The increase in the COMP neoepitope in the synovial fluid from horses with acute lameness suggests that this neoepitope has the potential to be a unique candidate biomarker for the early molecular changes in articular cartilage associated with OA.


Phase pattern denoising using a regularized cost function with complex-valued Markov random fields based on a discrete model.
We present a simple and effective method for denoising phase patterns based on a discrete model. The proposed filtering method transforms the image denoising problem to solving the energy diffusion problem of a system with complex-valued fields. We establish an appropriate cost function that uses the discrete form of complex-valued Markov random fields. The attractiveness of the proposed filtering method includes three points: the first is that the filtering process can be easily implemented using an iterative method, the second is that 2π phase jumps are well preserved, and the third is its little computational effort. The performance of the proposed method is demonstrated by simulated and experimentally obtained phase patterns.


Comparison of Outcome of Coronary Artery Bypass Grafting Versus Drug-Eluting Stent Implantation for Non-ST-Elevation Acute Coronary Syndrome.
There is limited data comparing effectiveness of coronary artery bypass grafting (CABG) versus percutaneous coronary intervention (PCI) with drug-eluting stents in patients with non-ST-elevation acute coronary syndromes (NSTE-ACS). We compared the long-term outcomes of the 2 revascularization strategies in 1,246 patients presented with NSTE-ACS for left main or multivessel coronary artery disease. Data were pooled from the Randomized Comparison of Coronary Artery Bypass Surgery and Everolimus-Eluting Stent Implantation in the Treatment of Patients with Multivessel Coronary Artery Disease (BEST) trial, the Premier of Randomized Comparison of Bypass Surgery versus Angioplasty Using Sirolimus-Eluting Stent in Patients with Left Main Coronary Artery Disease (PRECOMBAT) trial, and the Synergy between PCI with Taxus and Cardiac Surgery (SYNTAX) trial. The primary outcome was a composite of death from any causes, myocardial infarction, or stroke. The baseline characteristics were similar between the 2 study groups. During the median follow-up of 60 months, the rate of the primary outcome was significantly lower with CABG than with PCI (hazard ratio [HR] 0.74; 95% confidence interval [CI] 0.56 to 0.98; p = 0.036). This difference was mainly attributed to a significant reduction in the rate of myocardial infarction (HR 0.50; 95% CI 0.31 to 0.82, p = 0.006). The superiority of CABG over PCI was consistent across the major subgroups. The individual risks of death from any causes or stroke were not different between the 2 groups. In contrast, the rate of repeat revascularization was significantly lower in the CABG group than in the PCI group (HR 0.56; 95% CI 0.41 to 0.75, p <0.001). In this study, among patients with NSTE-ACS for left main or multivessel coronary artery disease, CABG significantly reduces the risk of death from any causes, myocardial infarction, or stroke compared with PCI with drug-eluting stents.


Is manual reticulocyte count a reliable option for under resourced countries?
To establish the credibility of manual reticulocyte counts without compromising the quality of care, and to evaluate the degree of acceptability of manual reticulocyte counts in terms of accuracy and cost effectiveness in comparison with two automated haematology analyzers. Visual reticulocyte enumeration was evaluated for comparability, within-batch precision and costing with respect to Coulter STKS and Gen S haematology analyzers. The results of reticulocyte estimation for 80 samples as obtained by 3 modes were correlated using Pearson's correlation coefficient (r) which were computed as 0.884, 0.875, and 0.793 for manual-Gen.S, Gen.SSTKS and manual-STKS respectively thus showing positive association of these results. STKS had the CV of 10.4% and was more precise compared to Coulter Gen.S (CV = 11.6%) while manual counts showed the least precision with a CV of 19.8%. The cost per test was calculated to be $0.11 for manual technique in contrast to $0.45 for Gen S and $1.09 for STKS. Visual counting of reticulocytes can be used as a reliable tool for estimating reticulocytes in resource strained countries as it is not only cost effective but can also efficiently discriminate between high and low reticulocyte ranges which are required for sound clinical judgment.


Metabolism and pharmacokinetics of 1,3-dinitrobenzene in the rat and the hamster.
Recent studies have documented that the Sprague-Dawley rat is markedly more sensitive than the Golden Syrian hamster to 1,3-dinitrobenzene (1,3-DNB)-induced testicular toxicity and methemoglobinemia. The present studies have investigated the possibility that differences in 1,3-DNB metabolism and pharmacokinetics could explain this species difference. [14C]1,3-DNB (25 mg/kg, ip) was administered to both species at a dose known to induce a testicular lesion in the rat and levels of 1,3-DNB and its metabolites were measured in blood and urine. Elimination of 1,3-DNB from the blood was initially rapid, followed by a second, much slower phase. Peak blood levels of 1,3-DNB were very different between the two species, with the hamster reaching levels only one-half those found in the rat (46.3 vs. 99.5 nmol/ml, respectively). Administration of a 50-mg/kg dose to the hamster resulted in 1,3-DNB blood levels which were similar to those found in the rat at 25 mg/kg. Since no obvious testicular toxicity is apparent in the hamster even at the higher dose level, a direct effect of 1,3-DNB on the testes seems unlikely, although it is possible that hamster cells inherently lack sensitivity to toxicity. Other major differences between the two species were a more rapid initial elimination rate and much higher blood levels of nitroaniline in the rat. Analysis of urinary metabolites revealed that the rat excreted more unconjugated metabolites and less phenolic metabolites compared with the hamster. Overall, the data indicate that the capacity to form reductive metabolites may play an important role in susceptibility to toxicity.


Association Between Atrial Fibrillation and Dementia: A Meta-Analysis.
Background: A potential evidence from previous epidemiological studies remains conflicting findings regarding the association between atrial fibrillation (AF) and dementia risk. We, therefore, carried out a meta-analysis of relevant studies to investigate the magnitude of the association between AF and dementia risk. Methods: We performed a systematic literature search of PubMed, EMBASE, and Google Scholar for potential studies between January 1, 1990, and December 31, 2018, with no restriction on the publication language. All potential studies were independently assessed by two reviewers. We only included observational studies that calculated the odds ratio (OR)/hazards ratio (HR) for dementia associated with atrial fibrillation. We first assessed the heterogeneity among study-specific HRs using the Q statistic and I 2 statistic. We then used the random-effects model to obtain the overall HR and its 95% CI for all studies. We also tested and corrected for publication bias by funnel plot-based methods. The quality of each study was assessed with the Newcastle Ottawa Scale. Results: A total of 16 studies with 2,415,356 individuals, and approximately 200,653 cases of incidence dementia were included in this study. Patients with AF had a greater risk of incidence dementia than those without AF (random-effect hazard ratio HR: 1.36, 95% CI: 1.23-1.51, p < 0.0001; I 2 = 83.58). Funnel plot and Egger test did not reveal significant publication bias. However, limitations of the study included high heterogeneity and varying degrees of confounder adjustment across individual studies. Conclusion: This study serves as added evidence supporting the hypothesis that AF is associated with an increased risk of dementia. More studies are needed to establish whether optimal treatment of AF can reduce or mitigate the risk of dementia.


Sudden death as presenting symptom caused by cardiac primary multicentric left ventricle rhabdomyoma, in an 11-month-old baby. An immunohistochemical study.
This case report describes a sudden cardiac death in an apparent healthy 11-month-old infant caused by a multifocal cardiac rhabdomyoma. Parents reported that a few days before the child had fallen to the ground getting a little superficial injury to the scalp. The authors hypothesize that it may have been a transient loss of consciousness episode caused by the cardiac tumour. After the gross examination, histological investigation supported by immunohistochemical analysis using antibody anti-Myoglobin, Actin, Vimentin, Desmin, CD34, S-100, Ki-67 was carried out for the diagnosis. Death was attributed to a multifocal cardiac rhabdomyoma, a benign tumour of striated muscle, which has been completely asymptomatic. In particular, one mass filled the entire posterior wall of the left ventricle. The insidious development of benign cardiac tumours also in infants and children is outlined, focusing on the responsible mechanisms of sudden death in such cases and providing a reference for additional study on these subjects. Virtual slides: The virtual slide(s) for this article can be found here:http://www.diagnosticpathology.diagnomx.eu/vs/7163626988365078.


Comparison of printed glycan array, suspension array and ELISA in the detection of human anti-glycan antibodies.
Anti-glycan antibodies represent a vast and yet insufficiently investigated subpopulation of naturally occurring and adaptive antibodies in humans. Recently, a variety of glycan-based microarrays emerged, allowing high-throughput profiling of a large repertoire of antibodies. As there are no direct approaches for comparison and evaluation of multi-glycan assays we compared three glycan-based immunoassays, namely printed glycan array (PGA), fluorescent microsphere-based suspension array (SA) and ELISA for their efficacy and selectivity in profiling anti-glycan antibodies in a cohort of 48 patients with and without ovarian cancer. The ABO blood group glycan antigens were selected as well recognized ligands for sensitivity and specificity assessments. As another ligand we selected P(1), a member of the P blood group system recently identified by PGA as a potential ovarian cancer biomarker. All three glyco-immunoassays reflected the known ABO blood groups with high performance. In contrast, anti-P(1) antibody binding profiles displayed much lower concordance. Whilst anti-P(1) antibody levels between benign controls and ovarian cancer patients were significantly discriminated using PGA (p=0.004), we got only similar results using SA (p=0.03) but not for ELISA. Our findings demonstrate that whilst assays were largely positively correlated, each presents unique characteristic features and should be validated by an independent patient cohort rather than another array technique. The variety between methods presumably reflects the differences in glycan presentation and the antigen/antibody ratio, assay conditions and detection technique. This indicates that the glycan-antibody interaction of interest has to guide the assay selection.


A Plea for MERSQI: The Medical Education Research Study Quality Instrument.
To describe the quality of educational scholarship presented at a large national conference of obstetrics and gynecology educators. We reviewed Council on Resident Education in Obstetrics and Gynecology-Association of Professors of Gynecology and Obstetrics annual meeting abstracts from 2015 and 2016, published as supplements to Obstetrics & Gynecology. For this uncontrolled observational study, abstracts were reviewed and scored using the Medical Education Research Study Quality Instrument (MERSQI). Comparisons between types of submissions (oral presentations or posters), origin of the report (academic or community), setting (undergraduate, graduate, postgraduate), and focus of the study (tool development or evaluation) were made. Abstracts from award-winning presentations and full manuscripts were compared with the remaining abstracts. One- and two-tailed Student t tests with a two-sample unequal variance (heteroscedastic) test were performed with a significance threshold of P≤.05. One hundred eighty-six abstracts and articles were available, with 101 posters and 77 oral presentations that could be scored in all six of the MERSQI domains. The average MERSQI score was 9.05 (±1.90) with scores ranging from 5 to 13.5 (median 9). Abstracts from full-text articles scored more than 1 point higher than other abstracts (10.2 compared with 9.0, P<.001, Cohen's d=0.72). Statistically significant smaller magnitude differences were found comparing tool development with evaluation, academic with community studies, and for award with nonaward winners. No differences were found comparing oral and poster presentations. The quality of educational scholarship presented at a national meeting of obstetrics and gynecology educators falls within the published range for other specialties. The MERSQI scoring system is a useful method for tracking and benchmarking the quality of medical education scholarship in obstetrics and gynecology.


Loss of tumorigenicity and increased immunogenicity induced by interleukin-10 gene transfer in B16 melanoma cells.
Because interleukin-10 (IL-10) has potent immunosuppressive and anti-inflammatory properties and is produced by some cancers, we hypothesized that its production might play a role in carcinogenesis by inhibiting adequate antitumoral immune responses. To test this hypothesis, retroviral vectors containing the IL-10 cDNA were generated and used to infect B16F1 melanoma cells that were injected subcutaneously in syngeneic mice. Surprisingly, IL-10 gene transfer resulted in a loss of tumorigenicity that was proportional to the amount of IL-10 secreted. Histological analysis showed massive area of necrosis of these tumor cells, with infiltration of polymorphic inflammatory cells. Parental cells simultaneously implanted had decreased tumorigenicity only when mixed with IL10-producing cells, but not when injected contralaterally, suggesting that their eradication is mediated mostly by a local phenomenon. Host T lymphocytes and natural killer (NK) cells were involved in this eradication because IL-10-producing cells grew in nude mice and in CD8+ or NK-depleted mice. Finally, mice injected with IL-10-secreting cells developed an antitumoral systemic immune response able to protect them against a subsequent challenge with parental cells. These results demonstrate that, in some settings, IL10 may have in vivo immunostimulating and proinflammatory properties that need to be considered in its therapeutic development.


Cohort Profile: The Precipitating Events Project (PEP Study).
The Precipitating Events Project (PEP Study) is an ongoing longitudinal study of 754 nondisabled community-living persons age 70 years or older who were members of a large health plan in greater New Haven, Connecticut, USA. The study was established to rigorously evaluate the epidemiology of disability in older persons and to elucidate the role of intervening illnesses and injuries on the disabling process. Of the eligible members, 75.2% agreed to participate and were enrolled between March 1998 and October 1999. Participants have completed comprehensive home-based assessments at 18-month intervals and have been interviewed monthly over the phone with a completion rate of 99%. Detailed participant-level data on health care utilization are obtained annually through linkages with Medicare claims. Through June 2019, 702 (93.1%) participants have died after a median of 109 months, while 43 (5.7%) have dropped out of the study after a median of 27 months. Death certificates are available for all decedents. To date, 117 original reports have been published using data from the PEP Study, including many focusing on other high priority areas such as end of life, frailty, depressive symptoms, aging stereotypes, pain, sleep, and methodologic research. The PEP Study welcomes proposals to access data for meritorious analyses from qualified investigators.


First Report of Bacterial Leaf Spot of Pumpkin Caused by Xanthomonas cucurbitae in Georgia, United States.
In August 2012, a commercial pumpkin (Cucurbita maxima L. cv. Neon) field in Terrell County, GA, had a disease outbreak that caused severe symptoms on leaves and fruits. Leaves displayed small (2 to 3 mm), angular, water-soaked, yellow lesions while fruits had small (2 to 3 mm), sunken, circular, dry lesions. The field exhibited 40% disease incidence with observable symptoms on fruits. In severe cases, fruit rots were also observed. Symptomatic leaves and fruits were collected from 25 pumpkin plants and isolations were made on both nutrient agar and yeast extract-dextrose-CaCO3 (YDC) agar medium (1). Xanthomonad-like yellow colonies were observed on both agar plates and colonies appeared mucoid on YDC. Suspect bacteria were gram-negative, oxidase positive, hydrolyzed starch and esculin, formed pits on both crystal violet pectate and carboxymethyl cellulose media, but were indole negative and did not produce nitrites from nitrates. Bacterial isolates also produced hypersensitive reactions on tobacco when inoculated with a bacterial suspension of 1 × 108 CFU/ml. Identity of the isolates were identified as genus Xanthomonas by using primers RST2 (5'AGGCCCTGGAAGGTGCCCTGGA3') and RST3 (5'ATCGCACTGCGTACCGCGCGCGA3') in a conventional PCR assay, which produced an 840-bp band. The 16S rRNA gene of five isolates was amplified using universal primers fD1 and rD1 (3) and amplified products were sequenced and compared using BLAST in GenBank. The nucleotide sequences (1,200 bp) of the isolates matched Xanthomonas cucurbitae (GenBank Accession AB680438.1), X. campestris (HQ256868.1), X. campestris pv. campestris (NR074936.1), X. hortorum (AB775942.1), and X. campestris pv. raphani (CP002789.1) with 99% similarity. PCR amplification and sequencing of a housekeeping gene atpD (ATP synthase, 720 bp) showed 98% similarity with X. cucurbitae (HM568911.1). Since X. cucurbitae was not listed in the BIOLOG database (Biolog, Hayward, CA), substrate utilization tests for three pumpkin isolates were compared with utilization patterns of Xanthomonas groups using BIOLOG reported by Vauterin et al. (4). The isolates showed 94.7, 93.7, and 92.6% similarity to the reported metabolic profiles of X. campestris, X. cucurbitae, and X. hortorum, respectively, of Xanthomonas groups 15, 8, and 2. However, PCR assay with X. campestris- and X. raphani-specific primers (3) did not amplify the pumpkin isolates, indicating a closer relationship with X. cucurbitae. Spray inoculations of five bacterial isolates in suspensions containing 1 × 108 CFU/ml on 2-week-old pumpkin seedlings (cv. Lumina) (n = five seedlings/isolate/experiment) under greenhouse conditions of 30°C and 70% RH produced typical yellow leaf spot symptoms on 100% of the seedlings. Seedlings (n = 10) spray-inoculated with sterile water were asymptomatic. Reisolated bacterial colonies from symptomatic seedlings displayed similar characteristics to those described above. Further confirmation of bacterial identity was achieved by amplifying and sequencing the 16S rRNA gene, which showed 98 to 99% similarity to X cucurbitae accessions in GenBank. To our knowledge, this is the first report of X. cucurbitae on pumpkin in Georgia. As this bacterium is known to be seedborne, it is possible that the pathogen might have introduced through contaminated seeds. References: (1) N. W. Schaad et al. Laboratory Guide for the Identification of Plant Pathogenic Bacteria, third edition. APS Press. St. Paul, MN, 2001. (2) Y. Besancon et al. Biotechnol. Appl. Biochem. 20:131, 1994. (3) Leu et al. Plant Pathol. Bull. 19:137, 2010. (4) Vauterin et al. Int. J. Syst. Bacteriol. 45:472, 1995.


Pharmacogenetic assessment of tafenoquine efficacy in patients with Plasmodium vivax malaria.
Plasmodium vivax has the largest geographic range of human malaria species and is challenging to manage and eradicate due to its ability to establish a dormant liver stage, the hypnozoite, which can reactivate leading to relapse. Until recently, the only treatment approved to kill hypnozoites was the 8-aminoquinoline, primaquine, requiring daily treatment for 14 days. Tafenoquine, an 8-aminoquinoline single-dose treatment with activity against P. vivax hypnozoites, has recently been approved by the US Food and Drug Administration and Australian Therapeutic Goods Administration for the radical cure of P. vivax malaria in patients 16 years and older. We conducted an exploratory pharmacogenetic analysis (GSK Study 208099) to assess the role of host genome-wide variation on tafenoquine efficacy in patients with P. vivax malaria using data from three GSK clinical trials, GATHER and DETECTIVE Part 1 and Part 2. Recurrence-free efficacy at 6 and 4 months and time to recurrence up to 6 months postdosing were analyzed in 438 P. vivax malaria patients treated with tafenoquine. Among the approximately 10.6 million host genetic variants analyzed, two signals reached genome-wide significance (P value ≤ 5 × 10). rs62103056, and variants in a chromosome 12 intergenic region, were associated with recurrence-free efficacy at 6 and 4 months, respectively. Neither of the signals has an obvious biological rationale and would need replication in an independent population. This is the first genome-wide association study to evaluate genetic influence on response to tafenoquine in P. vivax malaria.


Milk--the promoter of chronic Western diseases.
Common chronic diseases of Western societies, such as coronary heart disease, diabetes mellitus, cancer, hypertension, obesity, dementia, and allergic diseases are significantly influenced by dietary habits. Cow's milk and dairy products are nutritional staples in most Western societies. Milk and dairy product consumption is recommended by most nutritional societies because of their beneficial effects for calcium uptake and bone mineralization and as a source of valuable protein. However, the adverse long-term effects of milk and milk protein consumption on human health have been neglected. A hypothesis is presented, showing for the first time that milk protein consumption is an essential adverse environmental factor promoting most chronic diseases of Western societies. Milk protein consumption induces postprandial hyperinsulinaemia and shifts the growth hormone/insulin-like growth factor-1 (IGF-1) axis to permanently increased IGF-1 serum levels. Insulin/IGF-1 signalling is involved in the regulation of fetal growth, T-cell maturation in the thymus, linear growth, pathogenesis of acne, atherosclerosis, diabetes mellitus, obesity, cancer and neurodegenerative diseases, thus affecting most chronic diseases of Western societies. Of special concern is the possibility that milk intake during pregnancy adversely affects the early fetal programming of the IGF-1 axis which will influence health risks later in life. An accumulated body of evidence for the adverse effects of cow's milk consumption from fetal life to childhood, adolescence, adulthood and senescence will be provided which strengthens the presented hypothesis.


Challenges and Opportunities towards the Development of Risk Assessment at the Consumer Phase in Developing Countries-The Case of Campylobacter Cross-Contamination during Handling of Raw Chicken in Two Middle Eastern Countries.
In many low- and middle-income countries, data limitations are a major challenge facing the development of food safety risk assessment. In the present study, a questionnaire data collection tool was designed with an emphasis on gathering specific data points required by a risk modeller for simulating a scenario of Campylobacter cross-contamination during handling of raw chicken meat at the consumer phase. The tool was tested in practice to support its value and applicability in settings where data limitations are a challenge. The study subjects were 450 consumers in two Middle Eastern settings: Alexandria in Egypt (n = 200) and Thi-Qar in Iraq (n = 250). The majority (78.5%) of respondents in Egypt opted for wet markets/live bird shops as their preferred source of chicken meat. In contrast, 59.6% of Iraqi respondents preferred to buy chicken meat from supermarkets. Added to that, 73.0% of consumers in Egypt and 56.8% of consumers in Iraq viewed the quality of frozen chicken as "inferior" to that of chicken from wet markets. Almost all respondents in both Egypt and Iraq shared the practice of washing chicken in water before cooking. The percentage of consumers who 'very frequently' or 'frequently' prepare chicken prior to making the salad was 32.5% and 55.2% in Egypt and Iraq, respectively. A sizeable proportion of respondents in Iraq (40.8%) reported that they did not consider washing their hands with soapy water after touching raw chicken and preparing a salad in their home kitchen. Finally, 28.8% and 6.5% of respondents in Iraq and Egypt, respectively, indicated that they would not consider using a separate cutting board to avoid cross-contamination between raw chicken and salad. The data collection tool used in this study was designed in the first instance to match a conceptualised risk assessment framework, and that enabled the simultaneous collection of data points on consumption frequency, serving sizes, purchasing patterns, retail chain diversity and food handling practices. Results from such study design could be used for future development of a quantitative risk assessment model and to support food safety promotion efforts for domestic consumers in two of the most populated Middle Eastern countries.


Transcranial direct current stimulation (tDCS) in a realistic head model.
Distributions of current produced by transcranial direct current stimulation (tDCS) in humans were predicted by a finite-element model representing several individual and collective refinements over prior efforts. A model of the entire human head and brain was made using a finely meshed (1.1x1.1x1.4mm(3) voxel) tissue dataset derived from the MRI data set of a normal human brain. The conductivities of ten tissues were simulated (bone, scalp, blood, CSF, muscle, white matter, gray matter, sclera, fat, and cartilage). We then modeled the effect of placing a "stimulating" electrode with a saline-like conductivity over F3, and a similar "reference" electrode over a right supraorbital (RS) location, as well as the complements of these locations, to compare expectations derived from the simulation with experimental data also using these locations in terms of the presence or absence of subjective and objective effects. The sensitivity of the results to changes in conductivity values were examined by varying white matter conductivity over a factor of ten. Our simulations established that high current densities were found directly under the stimulating and reference electrodes, but values of the same order of magnitude occurred in other structures, and many areas of the brain that might be behaviorally active were also subjected to what may be substantial amounts of current. The modeling also suggests that more targeted stimulations might be achieved by different electrode topologies.


Determining Atomistic SAXS Models of Tri-Ubiquitin Chains from Bayesian Analysis of Accelerated Molecular Dynamics Simulations.
Small-angle X-ray scattering (SAXS) has become an increasingly popular technique for characterizing the solution ensemble of flexible biomolecules. However, data resulting from SAXS is typically low-dimensional and is therefore difficult to interpret without additional structural knowledge. In theory, molecular dynamics (MD) trajectories can provide this information, but conventional simulations rarely sample the complete ensemble. Here, we demonstrate that accelerated MD simulations can be used to produce higher quality models in shorter time scales than standard simulations, and we present an iterative Bayesian Monte Carlo method that is able to identify multistate ensembles without overfitting. This methodology is applied to several ubiquitin trimers to demonstrate the effect of linkage type on the solution states of the signaling protein. We observe that the linkage site directly affects the solution flexibility of the trimer and theorize that this difference in plasticity contributes to their disparate roles in vivo.


An open label, randomized clinical trial to compare the tolerability and efficacy of ivermectin plus diethylcarbamazine and albendazole vs. diethylcarbamazine plus albendazole for treatment of brugian filariasis in Indonesia.
Improved treatments for lymphatic filariasis (LF) could accelerate the global elimination program for this disease. A triple drug combination of the anti-filarial drugs ivermectin, diethylcarbamazine (DEC) and albendazole (IDA) has been shown to be safe and effective for achieving sustained clearance of microfilariae (Mf) of the filarial parasite Wuchereria bancrofti from human blood. However, the triple drug combination has not been previously been evaluated for treatment of brugian filariasis, which accounts for about 10% of the global LF burden. This hospital-based clinical trial compared the safety and efficacy of IDA with that of the standard treatment (DEC plus albendazole, DA) in persons with Brugia timori infections on Sumba island, Indonesia. Fifty-five asymptomatic persons with B. timori Mf were treated with either a single oral dose of IDA (28 subjects) or with DEC plus albendazole (DA, 27 subjects). Participants were actively monitored for adverse events (AE) for two days after treatment by nurses and physicians who were masked regarding treatment assignments. Passive monitoring was performed by clinical teams that visited participant's home villages for an additional five days. Microfilaremia was assessed by membrane filtration of 1 ml night blood at baseline, at 24h and one year after treatment. IDA was more effective than DA for completely clearing Mf at 24 hours (25/28, 89% vs. 8/27, 30%, P < 0.001). By 12 months after treatment, only one of 27 IDA recipients had Mf in their blood (4%) vs. 10 of 25 (40%) in persons treated with DA (P = 0.002). Approximately 90% of participants had antibodies to recombinant filarial antigen BmR1 at baseline. Antibody prevalence decreased to approximately 30% in both treatment groups at 12 months. About 45% of persons in both treatment groups experienced AE such as fever, muscle aches, lower back, joint and abdominal pain. These were mostly mild and most common during the first two days after treatment. No participant experienced a severe or serious AE. This study showed that IDA was well-tolerated and significantly more effective for clearing B. timori Mf from the blood than DA. Larger studies should be performed to further assess the safety and efficacy of IDA as a mass drug administration regimen to eliminate brugian filariasis. Trial Registration: NCT02899936.


Low or high doses of cefquinome targeting low or high bacterial inocula cure Klebsiella pneumoniae lung infections but differentially impact the levels of antibiotic resistance in fecal flora.
The combination of efficacious treatment against bacterial infections and mitigation of antibiotic resistance amplification in gut microbiota is a major challenge for antimicrobial therapy in food-producing animals. In rats, we evaluated the impact of cefquinome, a fourth-generation cephalosporin, on both Klebsiella pneumoniae lung infection and intestinal flora harboring CTX-M-producing Enterobacteriaceae. Germfree rats received a fecal flora specimen from specific-pathogen-free pigs, to which a CTX-M-producing Escherichia coli strain had been added. K. pneumoniae cells were inoculated in the lungs of these gnotobiotic rats by using either a low (10(5) CFU) or a high (10(9) CFU) inoculum. Without treatment, all animals infected with the low or high K. pneumoniae inoculum developed pneumonia and died before 120 h postchallenge. In the treated groups, the low-inoculum rats received a 4-day treatment of 5 mg/kg of body weight cefquinome beginning at 24 h postchallenge (prepatent phase of the disease), and the high-inoculum rats received a 4-day treatment of 50 mg/kg cefquinome beginning when the animals expressed clinical signs of infection (patent phase of the disease). The dose of 50 mg/kg targeting the high K. pneumoniae inoculum cured all the treated rats and resulted in a massive amplification of CTX-M-producing Enterobacteriaceae. A dose of 5 mg/kg targeting the low K. pneumoniae inoculum cured all the rats and averted an outbreak of clinical disease, all without any amplification of CTX-M-producing Enterobacteriaceae. These findings might have implications for the development of new antimicrobial treatment strategies that ensure a cure for bacterial infections while avoiding the amplification of resistance genes of human concern in the gut microbiota of food-producing animals.


Prognostic factors of perioperative FLOT regimen in operable gastric and gastroesophageal junction tumors: real-life data (Turkish Oncology Group).
Perioperative FLOT regimen is a standard of care in locally advanced operable gastric and GEJ adenocarcinoma. We aimed to determine the efficacy, prognostic factors of perioperative FLOT chemotherapy in real-life gastric and GEJ tumors. The data of patients who were treated with perioperative FLOT chemotherapy were retrospectively analyzed from 34 different oncology centers in Turkey. Baseline clinical and demographic characteristics, pretreatment laboratory values, histological and molecular characteristics were recorded. A total of 441 patients were included in the study. The median of age our study population was 60 years. The majority of patients with radiological staging were cT3-4N(+) (89.9%, n = 338). After median 13.5 months (IQR: 8.5-20.5) follow-up, the median overall survival was NR (95% CI, NR to NR), and median disease free survival was 22.9 (95% CI, 18.6 to 27.3) months. The estimated overall survival at 24 months was 62%. Complete pathological response (pCR) and near pCR was achieved in 23.8% of all patients. Patients with lower NLR or PLR have significantly longer median OS (p = 0.007 and p = 0.033, respectively), and patients with lower NLR have significantly longer median DFS (p = 0.039), but PLR level did not affect DFS (p = 0.062). The OS and DFS of patients with better ECOG performance scores and those who could receive FLOT as adjuvant chemotherapy instead of other regimens were found to be better. NLR was found to be independent prognostic factor for OS in the multivariant analysis. At least one adverse event reported in 57.6% of the patients and grade 3-4 toxicity was seen in 23.6% patients. Real-life perioperative FLOT regimen in operable gastric and GEJ tumors showed similar oncologic outcomes compared to clinical trials. Better performance status, receiving adjuvant chemotherapy as same regimen, low grade and low NLR and PLR improved outcomes in real-life. However, in multivariate analysis, only NLR affected OS.


Analysis of cytotoxicity induced by proinflammatory cytokines in the human alveolar epithelial cell line A549.
Epithelial cell injury under hyperinflammatory conditions is critical in the development of septic acute lung injury (ALI). The aim of the present study is to analyze the cytotoxic effects of a mixture of proinflammatory cytokines in the human alveolar epithelial cell line A549. The cytotoxicity of proinflammatory cytokines were assessed in A549 cells by measuring lactate dehydrogenase released into the culture medium and by crystal violet staining of surviving cells. Activation of the caspase-dependent apoptotic pathway was evaluated by monitoring cleavage of cytokeratin 18 by caspases using enzyme-liked immunosorbent assay (ELISA). To estimate the cytotoxic signaling pathways responsible for epithelial injury, agents with antiinflammatory or antioxidative properties were extensively screened for cytoprotective effects in the inflammation-associated epithelial injury model. The present study revealed that inflammatory cytokines exerted cytotoxicity in A549 cells. A mixture of interleukin-1beta (IL-1β), tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ), designated as cytomix, augmented cytotoxicity compared with each individual cytokine. Treatment with glucocorticoid (dexamethasone), tetracycline-derived antiinflammatory antibiotics (minocycline or doxycycline), angiotensin II receptor blockers (losartan or telmisartan), or antioxidants (dimethyl sulfoxide, catalase) attenuated cytomix-induced cytotoxicity, including caspase activation. These results implied that inflammatory cytokines alone could cause alveolar epithelial injury in the pathophysiology of septic ALI. Caspase-dependent apoptosis was speculated to be one mechanism responsible for the cytokine-induced cytotoxicity. Agents with antiinflammatory or antioxidative properties such as glucocorticoid, tetracycline-derived antibiotics, angiotensin II receptor blockers, or direct antioxidants showed substantial effect in attenuating cytokine-induced cytotoxicity and may be candidates for treatment options.


Impact of two oral doses of 100,000 IU of vitamin D3 in preschoolers with viral-induced asthma: a pilot randomised controlled trial.
New evidence supports the use of supplemental vitamin D in the prevention of exacerbation of asthma; however, the optimal posology to sufficiently raise serum levels while maximising adherence is unclear. The objective was to ascertain the efficacy of high-dose vitamin D3 in increasing serum vitamin D in preschoolers with asthma and provide preliminary data on safety and efficacy outcomes. We conducted a 7-month, triple-blind, randomised, placebo-controlled, pilot trial of children aged 1-5 years with viral-induced asthma. Participants were allocated to receive two oral doses of 100,000 IU vitamin D3 (intervention) or identical placebo (control) 3.5 months apart, once in the fall and once in the winter. Serum 25-hydroxyvitamin D (25OHD) was measured by tandem mass spectrometry at baseline, 10 days, 3.5 months, 3.5 months + 10 days, and 7 months. The main outcome was the change in serum 25OHD from baseline (Δ25OHD) over time and at 3.5 and 7 months; other outcomes included the proportion of children with 25OHD ≥ 75 nmol/L, safety, and adverse event rates. Children (N = 47) were randomised (intervention, 23; control, 24) in the fall. There was a significant adjusted group difference in the Δ25OHD (95% confidence interval) of 57.8 (47.3, 68.4) nmol/L, p < 0.0001), with a time (p < 0.0001) and group*time interaction effect (p < 0.0001), in favour of the intervention. A significant group difference in the Δ25OHD was observed 10 days after the first (119.3 [105.8, 132.9] nmol/L) and second (100.1 [85.7, 114.6] nmol/L) bolus; it did not reach statistical significance at 3.5 and 7 months. At 3.5 and 7 months, respectively, 63% and 56% of the intervention group were vitamin D sufficient (≥ 75 nmol/L) compared to 39% and 36% of the control group. Hypercalciuria, all without hypercalcaemia, was observed in 8.7% of intervention and 10.3% of control samples at any time point. Exacerbations requiring rescue oral corticosteroids, which appear as a promising primary outcome, occurred at a rate of 0.87/child. Two oral boluses of 100,000 IU vitamin D3,once in the fall and once in the winter, rapidly, safely, and significantly raises overall serum vitamin D metabolites. However, it is sufficient to maintain 25OHD ≥ 75 nmol/L throughout 7 months in only slightly more than half of participants. ClinicalTrials.gov, NCT02197702 (23 072014). Registered on 23 July 2014.


Immediate and short-term effects of the combination of dry needling and percutaneous TENS on post-needling soreness in patients with chronic myofascial neck pain.
Dry needling (DN) and percutaneous electrical nerve stimulation (PENS) are widely used techniques in the treatment of myofascial pain. To investigate the immediate and short-term effects of the combination of DN and PENS compared to DN alone on the upper trapezius muscle. This is a 72-hour follow-up single-blinded randomized controlled trial. Sixty-two volunteer patients with chronic myofascial neck pain with active Myofascial Trigger Points (MTrPs) in the upper trapezius muscle were recruited. Randomization was performed, and 31 patients received DN treatment (DN group) and 31 received DN and PENS (DN+PENS group). The primary outcomes were neck disability index (NDI) and visual analog scale for pain for both post-needling soreness (PNS) and neck pain intensity (NPI). Pressure pain threshold (PPT) and cervical range of motion (CROM) were the secondary outcomes. We detected between-group differences in NPI and PNS in favor of the DN+PENS group immediately after treatment. No between-group differences in NDI were observed. PENS application after dry needling treatment is more effective than dry needling alone for decreasing soreness in the short term and improving neck pain intensity immediately in patients with myofascial chronic neck pain.


The novel protein suppressed in lung cancer down-regulated in lung cancer tissues retards cell proliferation and inhibits the oncokinase Aurora-A.
In an attempt to search for genes with abnormal expression in cancers, Suppressed in Lung Cancer (SLAN, also known as KIAA0256) is found underexpressed in human lung cancer tissues by quantitative real-time PCR (Q-RT-PCR). The study set out to characterize SLAN protein and explore its cellular functions. SLAN or its specific short hairpin RNA, full length or various deletion mutants were overexpressed in 293T or lung cancer cell lines, and cell proliferation, cell cycle, mitosis progression, and spindle configuration were surveyed. SLAN and its deletion mutants are localized to many subcellular locations such as endoplasmic reticulum (ER), nucleus, nucleolus, spindle pole and midbody, suggesting SLAN may function as a multifunctional protein. Overexpression of SLAN per se or its short hairpin RNAs (shRNAs) inhibits or accelerates cell proliferation through prolonging or shortening mitosis. Time-lapse microscopic recording reveals that cells overexpressing exogenous SLAN are arrested in mitosis or cannot undergo cytokinesis. SLAN 2-551 mutants drastically arrest cells in mitosis, where α- and γ-tubulin are disorganized. SLAN employs C-terminal to interact with Aurora-A, a key mitosis regulator and an oncogenic kinase associated with a wide range of human cancers. SLAN negatively regulates the activity of Aurora-A by directly inhibiting kinase activity in vitro or reducing the level of active Aurora-A in cells. SLAN is frequently reduced in lung cancer tissues overexpressing Aurora-A, arguing for the necessity to suppress SLAN during the Aurora-A-associated cancer formation. Taken together, we have identified a novel protein SLAN downregulated in lung caner, having multiple subcellular localization including spindle matrix and midbody, inhibiting cell proliferation and Aurora-A.


Growth of Filipino infants who differ in body proportions at birth.
Growth of infants who differ in body proportions at birth was followed during the first 12 months of life. The sample of 2,695 infants from Metropolitan Cebu, in the Central Philippines, was divided into five groups based on a cross tabulation of low-birth-weight(LBW) status (2,500 g or less) with low (<10th percentile), adequate (10th-90th percentile) or high (>90th percentile)Rohre's Index (RI). The groups exhibited significant differences in growth patterns. The importance of these groups, which reflect weight and body proportions of the infant at birth as a determinant of attained size at 2, 6, and 12 months of age, was evaluated by using multivariate techniques which took into consideration other factors known to affect growth. Results show that intergroup differences in growth patterns and attained size occur because of (1) differences in postnatal growth potential represented by weight and body proportions at birth, (2) differences in the important biological inputs that affect postnatal growth (such as feeding practices and morbidity), and (3) differences in the effects of inputs on growth. Infants who are LBW but well proportioned are most likely to remain small during the first year of life, while those with a low RI at birth are more capable of catch-up growth.


Development and disease of the photoreceptor cilium.
Primary cilia are microtubule-rich hair-like extensions protruding from the surface of most post-mitotic cells. They act as sensory organelles that help interpret various environmental cues. Mutations in genes encoding proteins involved in ciliogenesis or protein transport to the primary cilia lead to a wide variety of diseases commonly referred to as ciliopathies,which include primary ciliary dyskinesia, situs invertus, hydrocephalus, kidney diseases, respiratory diseases, and retinal degenerations. In the retina, the photoreceptor cells have a highly specialized primary cilium called the outer segment (OS), which is essential for photosensation. Development of the photoreceptor OS shares key regulatory mechanisms with ciliogenesis in other cell types. Accumulating evidence indicates that mutations that affect OS development and/or protein transport to the OS generally lead to photoreceptor degeneration, which can be accompanied by a range of other clinical manifestations due to the dysfunction of primary cilia in different cell types. Here, we review the general mechanisms regulating ciliogenesis, and present different examples of mutations affecting OS ciliogenesis and protein transport that lead to photoreceptor degeneration. Overall, we conclude that the genetic and molecular evidence accumulated in recent years suggest a clear link between the development and function of the primary cilium and various clinical conditions. Future studies aimed at uncovering the cellular and molecular mechanisms implicated in ciliogenesis in a wide variety of animal models should greatly increase our understanding of the pathophysiology of many human diseases, including retinal degenerations.


T cell interleukin-17 induces stromal cells to produce proinflammatory and hematopoietic cytokines.
Analysis of the cDNA encoding murine interleukin (IL) 17 (cytotoxic T lymphocyte associated antigen 8) predicted a secreted protein sharing 57% amino acid identity with the protein predicted from ORF13, an open reading frame of Herpesvirus saimiri. Here we report on the cloning of human IL-17 (hIL-17), the human counterpart of murine IL-17. hIL-17 is a glycoprotein of 155 amino acids secreted as an homodimer by activated memory CD4+ T cells. Although devoid of direct effects on cells of hematopoietic origin, hIL-17 and the product of its viral counterpart, ORF13, stimulate epithelial, endothelial, and fibroblastic cells to secrete cytokines such as IL-6, IL-8, and granulocyte-colony-stimulating factor, as well as prostaglandin E2. Furthermore, when cultured in the presence of hIL-17, fibroblasts could sustain the proliferation of CD34+ hematopoietic progenitors and their preferential maturation into neutrophils. These observations suggest that hIL-17 may constitute (a) an early initiator of the T cell-dependent inflammmatory reaction; and (b) an element of the cytokine network that bridges the immune system to hematopoiesis.


Structure and dynamics of minke whale surfacing patterns in the Gulf of St. Lawrence, Canada.
Animal behavioral patterns can help us understand physiological and ecological constraints on animals and its influence on fitness. The surfacing patterns of aquatic air-breathing mammals constitute a behavioral pattern that has evolved as a trade-off between the need to replenish oxygen stores at the surface and the need to conduct other activities underwater. This study aims to better understand the surfacing pattern of a marine top predator, the minke whale (Balaenoptera acutorostrata), by investigating how their dive duration and surfacing pattern changes across their activity range. Activities were classified into resting, traveling, surface feeding and foraging at depth. For each activity, we classified dives into short and long dives and then estimated the temporal dependence between dive types. We found that minke whales modified their surfacing pattern in an activity-specific manner, both by changing the expression of their dives (i.e. density distribution) and the temporal dependence (transition probability) between dive types. As the depth of the prey layer increased between activities, the surfacing pattern of foraging whales became increasingly structured, going from a pattern dominated by long dives, when feeding at the surface, to a pattern where isolated long dives were followed by an increasing number of breaths (i.e. short dives), when the whale was foraging at depth. A similar shift in surfacing pattern occurred when prey handling time (inferred from surface corralling maneuvers) increased for surface feeding whales. The surfacing pattern also differed between feeding and non-feeding whales. Resting whales did not structure their surfacing pattern, while traveling whales did, possibly as a way to minimize cost of transport. Our results also suggest that minke whales might balance their oxygen level over multiple, rather than single, dive cycles.


High performance of low band gap polymer-based ambipolar transistor using single-layer graphene electrodes.
Bottom-contact bottom-gate organic field-effect transistors (OFETs) are fabricated using a low band gap pDTTDPP-DT polymer as a channel material and single-layer graphene (SLG) or Au source/drain electrodes. The SLG-based ambipolar OFETs significantly outperform the Au-based ambipolar OFETs, and thermal annealing effectively improves the carrier mobilities of the pDTTDPP-DT films. The difference is attributed to the following facts: (i) the thermally annealed pDTTDPP-DT chains on the SLG assume more crystalline features with an edge-on orientation as compared to the polymer chains on the Au, (ii) the morphological features of the thermally annealed pDTTDPP-DT films on the SLG electrodes are closer to the features of those on the gate dielectric layer, and (iii) the SLG electrode provides a flatter, more hydrophobic surface that is favorable for the polymer crystallization than the Au. In addition, the preferred carrier transport in each electrode-based OFET is associated with the HOMO/LUMO alignment relative to the Fermi level of the employed electrode. All of these experimental results consistently explain why the carrier mobilities of the SLG-based OFET are more than 10 times higher than those of the Au-based OTFT. This work demonstrates the strong dependence of ambipolar carrier transport on the source/drain electrode and annealing temperature.


Humanitarian Struggle in Burma's Conflict Zones.
The Back Pack Health Worker Team (BPHWT), a community- based health organization, provides primary health care to ethnic people in conflict, remote, and internally displaced areas, in Burma (aka Myanmar), controlled by ethnic armed organizations fighting against the Burma government. Its services include both curative and preventative health care through a network of 1,425 health personnel including community health workers and village-embedded traditional birth attendants and village health workers. The BPHWT organizational and program model may prove useful to Special Operations medical actions in support of insurgent movements and conversely with a host nation's counterinsurgency strategies, which include the extension of its health services into areas that may be remote and/or inhabited by indigenous people and have insurgency potential. In the former respect, special attention is directed toward "humanitarian struggle" that uses health care as a weapon against the counterinsurgency strategies of a country's oppressive military.


Establishment of a method to detect microalbuminuria by measuring the total urinary protein-to-creatinine ratio in diabetic patients.
Diabetes and chronic kidney disease (CKD) which are risk facters of cardiovascular disease, are increasing global public health problems. Microalbuminuria is an early sign of progressive cardiovascular and renal disease in individuals with or without diabetes. Screening for microalbuminuria and early treatment are recommended for patients with increased cardiovascular and renal risk factors. However, the procedure used to measure urinary albumin is expensive. Alternatively, the measurement of total urinary protein is simple and inexpensive. Thus, we aimed to establish a method that could predict the presence of microalbuminuria by measuring the total protein-to-creatinine ratio. Spot urine samples were obtained from 150 patients with diabetes mellitus, and the total protein-to-creatinine ratio and the albumin-to-creatinine ratio (ACR) were measured. There was a significant positive correlation between the protein-to-creatinine ratio and the ACR (r = 0.95). The presence of albuminuria (both micro- and macroalbuminuria) could be predicted from the value of the protein-to-creatinine ratio in more than 90% of patients. A receiver-operating characteristic curve analysis revealed that the protein-to-creatinine ratio had a sensitivity and a specificity of 90.8% and 91.9%, respectively, for the detection of albuminuria and a cutoff value of 0.091 g/g creatinine. These results suggest that screening for microalbuminuria can be replaced by the detection of the protein-to-creatinine ratio, which may be cost-effective for patients with cardiovascular risks as well as for the general population.


Crossmodal interactions between olfactory and visual learning in Drosophila.
Different modalities of sensation interact in a synergistic or antagonistic manner during sensory perception, but whether there is also interaction during memory acquisition is largely unknown. In Drosophila reinforcement learning, we found that conditioning with concurrent visual and olfactory cues reduced the threshold for unimodal memory retrieval. Furthermore, bimodal preconditioning followed by unimodal conditioning with either a visual or olfactory cue led to crossmodal memory transfer. Crossmodal memory acquisition in Drosophila may contribute significantly to learning in a natural environment.


Tristetraprolin Is a Prognostic Biomarker for Poor Outcomes among Patients with Low-Grade Prostate Cancer.
Background: We studied the utility of the tumor suppressor Tristetraprolin (TTP, ZFP36) as a clinically relevant biomarker of aggressive disease in prostate cancer patients after radical prostatectomy (RP).Methods: TTP RNA expression was measured in an RP cohort of patients treated at Moffitt Cancer Center (MCC) and obtained from six publically available RP datasets with biochemical recurrence (BCR; total n = 1,394) and/or metastatic outcome data (total n = 1,222). TTP protein expression was measured by immunohistochemistry in a tissue microarray of 153 MCC RP samples. The time to BCR or metastasis based on TTP RNA or protein levels was calculated using the Kaplan-Meier analysis. Univariable and multivariable Cox proportional hazard models were performed on multiple cohorts to evaluate if TTP is a clinically relevant biomarker and to assess if TTP improves upon the Cancer of the Prostate Risk Assessment postsurgical (CAPRA-S) score for predicting clinical outcomes.Results: In all of the RP patient cohorts, prostate cancer with low TTP RNA or protein levels had decreased time to BCR or metastasis versus TTP-high tumors. Further, the decreased time to BCR in TTP-low prostate cancer was more pronounced in low-grade tumors. Finally, pooled survival analysis suggests that TTP RNA expression provides independent information beyond CAPRA-S to predict BCR.Conclusions: TTP is a promising prostate cancer biomarker for predicting which RP patients will have poor outcomes, especially for low-grade prostate cancer patients.Impact: This study suggests that TTP RNA expression can be used to enhance the accuracy of CAPRA-S to predict outcomes in patients treated with RP. Cancer Epidemiol Biomarkers Prev; 27(11); 1376-83. ©2018 AACR.


A novel method to suppress the dispersal of Japanese cedar pollen by inducing morphologic changes with weak alkaline solutions.
Inhalation of airborne pollen causes irritative symptoms in humans, known as pollinosis. The changing global climate and increased pollution contribute to enhance the release of pollen, thereby increasing the number of people suffering from allergies. We examined the effect of spraying weak alkaline solutions onto cedar trees, the main allergenic culprit in Japan, on pollen release. Weak alkaline solutions were sprayed onto Japanese cedar blossoms to disrupt the external walls of the pollen, and to induce swelling of the cytosolic components containing the nucleus. This morphologic change of the pollen grains depended on the pH of the suspending solution, with a threshold pH of near 7.5. As the breakdown of the external walls and swelling of the cytosolic components are inhibited by high osmolarity, the influx of water triggered the morphologic changes. Weak alkaline solutions sprayed onto cedar blossoms decreased the amount of pollen released from the anthers in a pH dependent manner. The addition of detergent to the sodium bicarbonate solution facilitated this effect on cedar pollen release. We suggest that spraying cedar and cypress forests with a weak alkaline solution might prevent the scattering of pollen that causes allergies in humans.


Identification of human specific gene duplications relative to other primates by array CGH and quantitative PCR.
In order to identify human lineage specific (HLS) copy number differences (CNDs) compared to other primates, we performed pair wise comparisons (human vs. chimpanzee, gorilla and orangutan) by using cDNA array comparative genomic hybridization (CGH). A set of 23 genes with HLS duplications were identified, as well as other lineage differences in gene copy number specific of chimpanzee, gorilla and orangutan. Each species has gained more copies of specific genes rather than losing gene copies. Eleven of the 23 genes have only been observed to have undergone HLS duplication in Fortna et al. (2004) and in the present study. Then, seven of these 11 genes were analyzed by quantitative PCR in chimpanzee, gorilla and orangutan, as well as in other six primate species (Hylobates lar, Cercopithecus aethiops, Papio hamadryas, Macaca mulatta, Lagothrix lagothricha, and Saimiri sciureus). Six genes confirmed array CGH data, and four of them appeared to have bona fide HLS duplications (ABCB10, E2F6, CDH12, and TDG genes). We propose that these gene duplications have a potential to contribute to specific human phenotypes.


Bactericidal activity of 3D-printed hydrogel dressing loaded with gallium maltolate.
Chronic wounds are projected to reach epidemic proportions worldwide because of the aging population and the increasing incidence of diabetes. Despite extensive research, infection remains one of the leading sources of complications in chronic wounds, resulting in improper healing, biofilm formation, and lower extremity amputation. To address the limitations of standard treatments, we have developed a hydrogel wound dressing with self-tuning moisture control that incorporates a novel antimicrobial agent to eliminate and prevent infection. 3D-printing of a hydrogel dressing with dual porosity resulted in a new dressing with greater flexibility, increased water uptake, and more rapid swelling than bulk hydrogel dressings. Additionally, gallium maltolate (GaM) was incorporated into the dressing to investigate the efficacy of this antimicrobial agent. Loading profiles, release kinetics, and the bactericidal activity against Staphylococcus aureus (including methicillin-resistant Staphylococcus aureus) of GaM were investigated in vitro to identify target profiles that supported infection control. Finally, GaM-loaded hydrogel dressings were evaluated in vivo, utilizing a murine splinted-wound model that was inoculated with S. aureus. In comparison to an untreated control, GaM dressings markedly reduced the wound bacterial load without compromising wound closure rates. Overall, this work demonstrates the utility of a 3D-printed hydrogel dressing as an antimicrobial dressing to control infection in chronic wounds.


Cardiac MRI in Small Animals.
Cardiac magnetic resonance (MR) imaging of mice is a valuable tool for the precise in vivo diagnosis and prognosis of heart defects. This detailed protocol describes the method of cardiac MR imaging in mice step by step. A series of MR images captures the contractile function of the mouse heart and post-processing of the image data yields morphometric parameters (myocardial mass, myocardial wall thickness, ventricular end-systolic and end-diastolic volume) as well as functional parameters (stroke volume and ejection fraction). This protocol may also serve as a starting point for MR imaging of rats, by using larger image dimensions (field-of-view) and MR hardware suitable for larger animals.


Free-breathing contrast-enhanced multiphase MRI of the liver using a combination of compressed sensing, parallel imaging, and golden-angle radial sampling.
The objectives of this study were to develop a new method for free-breathing contrast-enhanced multiphase liver magnetic resonance imaging (MRI) using a combination of compressed sensing, parallel imaging, and radial k-space sampling and to demonstrate the feasibility of this method by performing image quality comparison with breath-hold cartesian T1-weighted (conventional) postcontrast acquisitions in healthy participants. This Health Insurance Portability and Accountability Act-compliant prospective study received approval from the institutional review board. Eight participants underwent 3 separate contrast-enhanced fat-saturated T1-weighted gradient-echo MRI examinations with matching imaging parameters: conventional breath-hold examination with cartesian k-space sampling volumetric interpolate breath hold examination (BH-VIBE) and free-breathing acquisitions with interleaved angle-bisection and continuous golden-angle radial sampling schemes. Interleaved angle-bisection and golden-angle data from each 100 consecutive spokes were reconstructed using a combination of compressed sensing and parallel imaging (interleaved-angle radial sparse parallel [IARASP] and golden-angle radial sparse parallel [GRASP]) to generate multiple postcontrast phases.Arterial- and venous-phase BH-VIBE, IARASP, and GRASP reconstructions were evaluated by 2 radiologists in a blinded fashion. The readers independently assessed quality of enhancement (QE), overall image quality (IQ), and other parameters of image quality on a 5-point scale, with the highest score indicating the most desirable examination. Mixed model analysis of variance was used to compare each measure of image quality. Images of BH-VIBE and GRASP had significantly higher QE and IQ values compared with IARASP for both phases (P < 0.05). The differences in QE between BH-VIBE and GRASP for the arterial and venous phases were not significant (P > 0.05). Although GRASP had lower IQ score compared with BH-VIBE for the arterial (3.9 vs 4.8; P < 0.0001) and venous (4.2 vs 4.8; P = 0.005) phases, GRASP received IQ scores of 3 or more in all participants, which was consistent with acceptable or better diagnostic image quality. Contrast-enhanced multiphase liver MRI of diagnostic quality can be performed during free breathing using a combination of compressed sensing, parallel imaging, and golden-angle radial sampling.


Relationship Between Early Childhood Non-Parental Childcare and Diet, Physical Activity, Sedentary Behaviour, and Sleep: A Systematic Review of Longitudinal Studies.
The rising prevalence of childhood obesity is a global public health concern. Evidence suggests that exposure to non-parental childcare before age six years is associated with development of obesity, diet, and activity behaviours (physical activity, sedentary behaviour, and sleep). However, findings are inconsistent and mostly from cross-sectional studies, making it difficult to identify the direction of causation in associations. This review identified and synthesised the published research on longitudinal associations between non-parental childcare during early childhood, diet, and activity behaviours. Seven databases were searched, and results were independently double-screened through title/abstract and full-text stages. Included studies were evaluated for risk of bias. Of the 18,793 references screened, 13 met eligibility criteria and were included in the review. These presented results on 89 tested childcare/outcome associations, 63 testing diet outcomes (59% null, remainder mixed), and 26 testing activity behaviour outcomes (85% null, remainder mixed). The scarce available literature indicates little and mixed evidence of a longitudinal association. This reflects a paucity of research, rather than clear evidence of no effect. There is an urgent need for studies investigating the longitudinal associations of non-parental childcare on diet and activity behaviours to assess potential lasting effects and mechanisms; whether and how effects vary by provider; and differences by intensity, duration, and population sub-groups.


A comparative study of three conservative treatments in patients with lumbar spinal stenosis: lumbar spinal stenosis with acupuncture and physical therapy study (LAP study).
Although the efficiency of conservative management for lumbar spinal stenosis (LSS) has been examined, different conservative management approaches have not been compared. We have performed the first comparative trial of three types of conservative management (medication with acetaminophen, exercise, and acupuncture) in Japanese patients with LSS. Patients with L5 root radiculopathy associated with LSS who visited our hospital for surgical treatment were enrolled between December 2011 and January 2014. In this open-label study, patients were assigned to three treatment groups (medication, exercise, acupuncture) according to the visit time. The primary outcomes were Zurich claudication questionnaire (ZCQ) scores before and after 4 weeks of treatment. Least square mean analysis was used to assess the following dependent variables in the treatment groups: changes in symptom severity and physical function scores of the ZCQ and the ZCQ score of patient's satisfaction after treatment. Thirty-eight, 40, and 41 patients were allocated to the medication, exercise, and acupuncture groups, respectively. No patient underwent surgical treatment during the study period. The symptom severity scores of the ZCQ improved significantly after treatment in the medication (p = 0.048), exercise (p = 0.003), and acupuncture (p = 0.04) groups. The physical function score improved significantly in the acupuncture group (p = 0.045) but not in the medication (p = 0.20) and exercise (p = 0.29) groups. The mean reduction in the ZCQ score for physical function was significantly greater for acupuncture than for exercise. The mean ZCQ score for treatment satisfaction was significantly greater for acupuncture than for medication. Acupuncture was significantly more effective than physical exercise according to the physical function score of the ZCQ and than medication according to the satisfaction score. The present study provides new important information that will aid decision making in LSS treatment. This study was registered with the UMIN Clinical Trials Registry ( UMIN000006957 ).


Endurance capacity, not body size, determines physical activity levels: role of skeletal muscle PEPCK.
Some people remain lean despite pressure to gain weight. Lean people tend to have high daily activity levels, but the source of this increased activity is unknown. We found that leanness cannot be accounted for by increased weight-corrected food intake in two different types of lean rats. As previously reported in lean people, we found that lean rats had higher daily activity levels; lean rats also expended more energy. These lean rats were developed through artificial selection for high aerobic endurance capacity. To test whether our findings extended to a human population, we measured endurance capacity using a VO(2max) treadmill test and daily activity in a group of non-exercising individuals. Similar to lean rats selectively bred for endurance capacity, our study revealed that people with higher VO(2max) also spent more time active throughout the day. Hence, endurance capacity may be the trait that underlies both physical activity levels and leanness. We identified one potential mechanism for the lean, active phenotype in rats, namely high levels of skeletal muscle PEPCK. Therefore, the lean phenotype is characterized by high endurance capacity and high activity and may stem from altered skeletal muscle energetics.


Influence of processing and storage of integral grape juice (Vitis labrusca L.) on its physical and chemical characteristics, cytotoxicity, and mutagenicity in vitro.
Integral grape juice is extracted from the grape through processes that allow the retention of their natural composition. However, due to the severity of some processes, fruit juices can undergo changes in their quality. The present study evaluated the cytotoxic and mutagenic effects of integral grape juice by a cytokinesis-blocked micronucleus assay in Rattus norvegicus hepatoma cells (HTC) in vitro. Vitis labrusca L. (variety Concord) were produced organically and by a conventional system, and their juice was extracted by a hot extraction process. The organic grapes were subjected to ultraviolet-type C radiation (UV-C). Experiments were performed after production and after 6 months in storage. Physicochemical analyses revealed that UV-C irradiation of organic grapes, the juice production process, and storage resulted in nutraceutical alterations. However, none of the juice concentrations were cytotoxic to HTC cells by the cytokinesis-blocked proliferation index results or were mutagenic, because the formation of micronucleated cells was not induced. In general, juice induced cell proliferation, possibly due to the presence of vitamins and sugar content (total soluble solid). The data increased the understanding of food technology and confirmed the quality and safety consumption of these juices.


The Aspergillus fumigatus transcription factor AceA is involved not only in Cu but also in Zn detoxification through regulating transporters CrpA and ZrcA.
Copper (Cu) and zinc (Zn) are essential nutrients for both pathogens and hosts; however, their excess accumulation is toxic for all cells. The Aspergillus transcription factor AceA has been identified having function for Cu detoxification through up-regulation of the expression of the P-type ATPase, CrpA. Here, we demonstrate that Aspergillus fumigatus CrpA is involved in both Cu and Zn detoxification and a putative metallothionein AfCrdA plays a major function in Cu detoxification, and a putative transporter AfZrcA has a dominant role in Zn detoxification, but all three members are transcriptionally dependent on AfAceA. Moreover, the Cys, RGHR, and KGRP motifs in the conserved N-terminus of AfAceA are essential, but not sufficient for AfAceA-mediated Cu and Zn tolerance. Our findings suggest that fungal pathogens have developed very precise systems with overlapping machinery to respond to the two different metal stressors. Meanwhile, there is separate specific machinery for Zn detoxification in response to high environmental Zn. Importantly, virulence testing demonstrated that the conserved Cu and Zn detoxification-related Cys residues in AfAceA have key roles in pathogenesis. Therefore, these findings will broaden the current understanding of the adaption of saprophytic fungi to Cu and Zn stress and their survival in hosts and other environmental niches.


Thiazolidinediones are partial agonists for the glucocorticoid receptor.
Although thiazolidinediones were designed as specific peroxisome proliferator-activated receptor (PPAR)-gamma-ligands, there is evidence for some off-target effects mediated by a non-PPARgamma mechanism. Previously we have shown that rosiglitazone has antiinflammatory actions not explicable by activation of PPARgamma,but possibly by the glucocorticoid receptor (GR). Rosiglitazone induces nuclear translocation both of GR-green fluorescent protein, and endogenous GR in HeLa and U20S cells but with slower kinetics than dexamethasone. Rosiglitazone also induces GR phosphorylation (Ser211), a GR ligand-binding-specific effect. Rosiglitazone drives luciferase expression from a simple glucocorticoid-response element containing reporter gene in a GR-dependent manner (EC50 4 microm), with a similar amplitude response to the partial GR agonist RU486. Rosiglitazone also inhibits dexamethasone-driven reporter gene activity (IC50 2.9 microm) in a similar fashion to RU486, suggesting partial agonist activity. Importantly we demonstrate a similar effect in PPARgamma-null cells, suggesting both GR dependence and PPARgamma independence. Rosiglitazone also activates a GAL4-GR chimera, driving a upstream activating sequence promoter, demonstrating DNA template sequence independence and furthermore enhanced steroid receptor coactivator-1-GR interaction, measured by a mammalian two-hybrid assay. Both ciglitazone and pioglitazone, structurally related to rosiglitazone, show similar effects on the GR. The antiproliferative effect of rosiglitazone is increased in U20S cells that overexpress GR, suggesting a biologically important GR-dependent component of rosiglitazone action. Rosiglitazone is a partial GR agonist, affecting GR activation and trafficking to influence engagement of target genes and affect cell function. This novel mode of action may explain some off-target effects observed in vivo. Additionally, antagonism of glucocorticoid action may contribute to the antidiabetic actions of rosiglitazone.


Activation of mouse peritoneal macrophages alters the structure and surface expression of protein-bound lactosaminoglycans.
We have begun to analyze and compare the surface carbohydrates present on populations of resident and activated mouse peritoneal macrophages. The activated macrophage populations studied include TG-elicited macrophages, BCG-activated macrophages, and resident macrophages cultured for 24 hr in the presence of lymphokines or heterologous serum. Analysis of glycopeptides generated by pronase digestion of surface glycoproteins labeled by the neuraminidase/galactose oxidase/NaB3H4 method indicates that the macrophage surface contains a class of high m.w. carbohydrates susceptible to degradation by endo-beta-galactosidase, lactosaminoglycans. These lactosaminoglycans are sialylated type 2 carbohydrates containing the repeating lactosamine disaccharide Gal beta 1-4GlcNAc as well as fucose residues. Macrophage activation was observed to markedly alter surface lactosaminoglycans. The alterations observed include 1) an increase in surface expression as determined by both an increase in neuraminidase/galactose oxidase/NaB3H4 labeling and by the ability of activated but not resident macrophages to bind I antibodies as assayed by indirect immunofluorescent surface staining, 2) the addition of alpha-galactose residues, and 3) an increase in GlcNAc beta 1-6Gal branching as indicated by an increased resistance to endo-beta-galactosidase degradation and by the ability of activated macrophages to bind I antibodies. These observations demonstrate that macrophage activation results in specific and substantial alterations in protein-bound surface carbohydrates.


Neprilysins: an evolutionarily conserved family of metalloproteases that play important roles in reproduction in Drosophila.
Members of the M13 class of metalloproteases have been implicated in diseases and in reproductive fitness. Nevertheless, their physiological role remains poorly understood. To obtain a tractable model with which to analyze this protein family's function, we characterized the gene family in Drosophila melanogaster and focused on reproductive phenotypes. The D. melanogaster genome contains 24 M13 class protease homologs, some of which are orthologs of human proteases, including neprilysin. Many are expressed in the reproductive tracts of either sex. Using RNAi we individually targeted the five Nep genes most closely related to vertebrate neprilysin, Nep1-5, to investigate their roles in reproduction. A reduction in Nep1, Nep2, or Nep4 expression in females reduced egg laying. Nep1 and Nep2 are required in the CNS and the spermathecae for wild-type fecundity. Females that are null for Nep2 also show defects as hosts of sperm competition as well as an increased rate of depletion for stored sperm. Furthermore, eggs laid by Nep2 mutant females are fertilized normally, but arrest early in embryonic development. In the male, only Nep1 was required to induce normal patterns of female egg laying. Reduction in the expression of Nep2-5 in the male did not cause any dramatic effects on reproductive fitness, which suggests that these genes are either nonessential for male fertility or perform redundant functions. Our results suggest that, consistent with the functions of neprilysins in mammals, these proteins are also required for reproduction in Drosophila, opening up this model system for further functional analysis of this protein class and their substrates.


Comparative genomics reveals mobile pathogenicity chromosomes in Fusarium.
Fusarium species are among the most important phytopathogenic and toxigenic fungi. To understand the molecular underpinnings of pathogenicity in the genus Fusarium, we compared the genomes of three phenotypically diverse species: Fusarium graminearum, Fusarium verticillioides and Fusarium oxysporum f. sp. lycopersici. Our analysis revealed lineage-specific (LS) genomic regions in F. oxysporum that include four entire chromosomes and account for more than one-quarter of the genome. LS regions are rich in transposons and genes with distinct evolutionary profiles but related to pathogenicity, indicative of horizontal acquisition. Experimentally, we demonstrate the transfer of two LS chromosomes between strains of F. oxysporum, converting a non-pathogenic strain into a pathogen. Transfer of LS chromosomes between otherwise genetically isolated strains explains the polyphyletic origin of host specificity and the emergence of new pathogenic lineages in F. oxysporum. These findings put the evolution of fungal pathogenicity into a new perspective.


The Use of Interferon Gamma Inducible Protein 10 as a Potential Biomarker in the Diagnosis of Latent Tuberculosis Infection in Uganda.
In the absence of a gold standard for the diagnosis of latent tuberculosis (TB) infection (LTBI), the current tests available for the diagnosis of LTBI are limited by their inability to differentiate between LTBI and active TB disease. We investigated IP-10 as a potential biomarker for LTBI among household contacts exposed to sputum positive active TB cases. Active TB cases and contacts were recruited into a cohort with six months' follow-up. Contacts were tested for LTBI using QuantiFERON®-TB Gold In-Tube (QFN) assay and the tuberculin skin test (TST). Baseline supernatants from the QFN assay of 237 contacts and 102 active TB cases were analysed for Mycobacterium tuberculosis (MTB) specific and mitogen specific IP-10 responses. Contacts with LTBI (QFN+TST+) had the highest MTB specific IP-10 responses at baseline, compared to uninfected contacts (QFN-TST-) p<0.0001; and active cases, p = 0.01. Using a cut-off of 8,239 pg/ml, MTB specific IP-10 was able to diagnose LTBI with a sensitivity of 87.1% (95% CI, 76.2-94.3) and specificity of 90.9% (95% CI, 81.3-96.6). MTB specific to mitogen specific IP-10 ratio was higher in HIV negative active TB cases, compared to HIV negative latently infected contacts, p = 0.0004. Concentrations of MTB specific IP-10 were higher in contacts with TST conversion (negative at baseline, positive at 6-months) than in those that were persistently TST negative, p = 0.001. IP-10 performed well in differentiating contacts with either latent or active TB from those who were uninfected but was not able to differentiate LTBI from active disease except when MTB specific to mitogen specific ratios were used in HIV negative adults. In addition, IP-10 had the potential to diagnose 'recent TB infection' in persons classified as having LTBI using the TST. Such individuals with strong IP-10 responses would likely benefit from chemoprophylaxis.


Somatotopy of the extrinsic laryngeal muscles in the human sensorimotor cortex.
The larynx is the major organ of vocalization. The intrinsic laryngeal muscles modify the internal shape of the larynx while the extrinsic laryngeal muscles move the entire larynx vertically in the airway. Previous neuroimaging research has established the somatotopic location of the intrinsic musculature of the larynx in the human motor cortex and showed it to be in an evolutionarily novel location compared to the homologous region in monkey cortex. In the current study, we attempted for the first time to determine the somatotopic localization of the extrinsic laryngeal musculature in humans. In a functional magnetic resonance imaging experiment, we had participants voluntarily move their larynx upward and/or downward in the airway in the absence of vocalization to engage the extrinsic laryngeal muscles or vocalize in the absence of vertical laryngeal movement to engage the intrinsic laryngeal muscles. Vertical movement of the larynx activated ventral pericentral sensorimotor cortex extending dorsally to overlap with the representation of the intrinsic laryngeal muscles. This pattern is a reversal from the somatotopy of the monkey, where the extrinsic laryngeal muscles are represented dorsally to the intrinsic laryngeal muscles.


Lactobacillus rhamnosus strain GG prevents enterohemorrhagic Escherichia coli O157:H7-induced changes in epithelial barrier function.
Enterohemorrhagic Escherichia coli (EHEC) O157:H7 intimately attaches to intestinal epithelial monolayers and produces attaching and effacing (A/E) lesions. In addition, EHEC infection causes disruptions of intercellular tight junctions, leading to clinical sequelae that include acute diarrhea, hemorrhagic colitis, and the hemolytic-uremic syndrome. Current therapy remains supportive since antibiotic therapy increases the risk of systemic complications. This study focused on the potential therapeutic effect of an alternative form of therapy, probiotic Lactobacillus rhamnosus strain GG, to attenuate EHEC-induced changes in paracellular permeability in polarized MDCK-I and T84 epithelial cell monolayers. Changes in epithelial cell morphology, electrical resistance, dextran permeability, and distribution and expression of claudin-1 and ZO-1 were assessed using phase-contrast, immunofluorescence, and transmission electron microscopy and macromolecular flux. This study demonstrated that pretreatment of polarized MDCK-I and T84 cells with the probiotic L. rhamnosus GG reduced morphological changes and diminished the number of A/E lesions induced in response to EHEC O157:H7 infection. With probiotic pretreatment there was corresponding attenuation of the EHEC-induced drop in electrical resistance and the increase in barrier permeability assays. In addition, L. rhamnosus GG protected epithelial monolayers against EHEC-induced redistribution of the claudin-1 and ZO-1 tight junction proteins. In contrast to the effects seen with the live probiotic, heat-inactivated L. rhamnosus GG had no effect on EHEC binding and A/E lesion formation or on disruption of the barrier function. Collectively, these findings provide in vitro evidence that treatment with the probiotic L. rhamnosus strain GG could prove to be an effective management treatment for preventing injury of the epithelial cell barrier induced by A/E bacterial enteropathogens.


Reversing growth deficiency in children: the effect of a community-based intervention.
The effect of a community health nursing intervention on children with growth deficiency (also called nonorganic failure to thrive or growth failure) was examined in a pretest and post-test experimental study. This study evaluated the impact of the intervention on growth quotients, children's diets, parent-child interaction, home environment, and mothers' perceived stress. The sample consisted of 39 children (ages 3 months to 3 years) with growth deficiency in weight or height for age, weight for height, or a decrease in growth across two percentiles. The children were enrolled in Special Supplemental Feeding Program for Women, Infants and Children (WIC) clinics in county health departments and were randomly assigned to experimental or control groups. After preliminary data were collected for the entire sample, a community-based intervention was administered to the experimental group during home visits. The intervention included education about nutrition and about parenting and community skills. Data collected after the intervention by a research assistant blind to group assignment indicated positive changes (P < or = .05) in the experimental group's growth quotients, home environments, and their mothers' perceived stress. This study supports the community health nursing practice of teaching nutrition and child care during home visits to families of children with growth deficiency.


A tale worth telling: the impact of the diagnosis experience on disclosure of genetic disorders.
Research suggests children with genetic disorders exhibit greater coping skills when they are aware of their condition and its heritability. While the experiences parents have at diagnosis may influence their decision to disclose the diagnosis to their children, there is little research into this communication. The aim of the current study was to examine the relationship between the diagnosis experience and the disclosure experience for parents of children with developmental disorders of a known genetic aetiology: parents of children with 22q11.2 deletion syndrome (22q11DS) were compared with a group of parents with children affected with other genetic diagnoses, with a similar age of diagnosis (e.g. fragile X syndrome) and a group where diagnosis generally occurs early (i.e. Down syndrome). The sample comprised 559 parents and caregivers of children with genetic developmental disorders, and an online survey was utilised. Items from the questionnaire were combined to create variables for diagnosis experience, parental disclosure experience, child's disclosure experience, and parental coping and self-efficacy. Across all groups parents reported that the diagnosis experience was negative and often accompanied by a lack of support and appropriate information. Sixty-eight per cent of those in the 22q11DS and 58.3% in the Similar Conditions groups had disclosed the diagnosis to their child, whereas only 32.7% of the Down syndrome group had. Eighty-six per cent of the Down syndrome group felt they had sufficient information to talk to their child compared with 44.1% of the Similar Conditions group and 32.6% of the 22q11DS group. Parents reported disclosing the diagnosis to their child because they did not want to create secrets; and that they considered the child's age when disclosing. In the 22q11DS and Similar Conditions groups, a poor diagnosis experience was significantly associated with negative parental disclosure experiences. In the Similar Conditions group, a poor diagnosis experience was also significantly associated with a more negative child disclosure experience. As expected this study highlights how difficult most parents find the diagnosis experience. Importantly, the data indicate that the personal experiences the parents have can have a long-term impact on how well they cope with telling their child about the diagnosis. It is important for clinicians to consider the long-term ramifications of the diagnosis experience and give the parents opportunities; through, for instance, psychoeducation to prepare for telling their child about the diagnosis. Further research is warranted to explore what type of information would be useful for parents to receive.


Tannin extracted from Sumac inhibits vascular smooth muscle cell migration.
Vascular smooth muscle cell (VSMC) migration is integral in the pathogenesis of atherosclerosis. Sumac (Rhus coriaria) berries are believed to have atheroprotective effects. Therefore, Sumac, which is a rich source of tannin antioxidants, was tested for its capacity to inhibit VSMC migratory activity. Tannin was extracted and purified from ground Sumac. Cultured rat carotid VSMCs were treated with different concentrations of tannin. After 10 days of tannin treatment, VSMC migratory activity in response to platelet-derived growth factor-BB was measured by transmembrane migration assay. An equal number of VSMCs was loaded on top of the inserts and at the bottom of the wells. After fixation and staining, cells migrating through the inserts and cells seeded at the bottom of the wells were counted. A significant reduction (62%) of VSMC migration was evident in tannin-treated cells. To rule out any possible toxicity and cell death, cells at the bottom of the wells were also counted. No difference between the tannin-treated group and the controls was observed in the number of cells seeded at the bottom of the wells. Our data suggest that tannin extracted from Sumac possesses potent antimigratory activity. Sumac may have potential for the prevention or treatment of atherosclerosis and its clinical manifestations. Further experiments, especially in vivo, are required to examine the atheroprotective effect of Sumac.


Measuring labor supply and demand shocks during COVID-19.
We measure labor demand and supply shocks at the sector level around the COVID-19 outbreak by estimating a Bayesian structural vector autoregression on monthly statistics of hours worked and real wages. Most sectors were subject to large negative labor supply and demand shocks in March and April 2020, with substantial heterogeneity in the size of shocks across sectors. Our estimates suggest that two-thirds of the drop in the aggregate growth rate of hours in March and April 2020 are attributable to labor supply. We validate our estimates of supply shocks by showing that they are correlated with sectoral measures of telework.


Acetabular Chondral Lesions in Hip Arthroscopy: Relationships Between Grade, Topography, and Demographics.
Hip pain remains a challenge given the multiple factors that can cause damage to the articular cartilage, such as traumatic injury, metabolic damage, and morphologic variations such as femoroacetabular impingement (FAI) and that can contribute to progression of osteoarthritis. However, a direct relationship between patient characteristics, the extent of acetabular chondral damage, and topologic characteristics of chondral lesions has not been established. To compare the grade of acetabular chondral damage, measured in terms of acetabular labrum articular disruption (ALAD) classification, to the size and position of the chondral lesions, matching patients' demographic factors such as age and body mass index (BMI). Cross-sectional study; Level of evidence, 3. This study included all hip arthroscopies performed by the corresponding author from August 7, 2008, to November 19, 2014, in which acetabular chondral lesions were intraoperatively identified and measured in terms of ALAD grade, clockface location, and size. Bivariate analyses and multiple logistic regression were used to identify the demographic factors, characteristics of the acetabular chondral lesion, and other anatomic characteristics that were related to the ALAD grade of the acetabular chondral lesion. Acetabular chondral lesions were measured in 1502 patients during the study period. Multivariate analysis showed that higher ALAD grade of acetabular chondral damage was significantly related to male sex, more advanced age, the area of the acetabular chondral lesion, anterior extension of the acetabular chondral lesion within the anterosuperior quadrant, labral detachment from the acetabular cartilage, and posterior extension of the labral tear. Higher grades of acetabular chondral damage were related to male sex, increased age, height, weight, BMI, and the size of the lesion. Chondral lesions were generally found in the anterosuperior region of the acetabulum, consistent with labral lesions and the weightbearing area of the acetabulum.


Endoscopic features of Helicobacter pylori induced gastritis.
It's still controversial whether certain macroscopic endoscopic features can be used to diagnose Helicobacter pylori (HP) related gastritis. The literature dealing with this subject is confusing, because of the lack of precise terminology, no large control trials, major discrepancies in interpretations of macroscopic changes and poor correlation of the macroscopic appearance and histological finding of gastritis. We conducted a prospective study of 208 dyspeptic patients, who underwent upper gastrointestinal endoscopies from February 1997 to June 1997. Only those patients who had either normal looking gastric mucosa or macroscopic gastritis were included in the study. Endoscopically normal looking mucosa was seen in 67 patients (65.6%), erythematous gastritis in 51 (74%), mosaic appearance in 18 (88%), erosive gastritis in 14 (85%), nodular gastritis in 17 (94%), atrophic gastritis in 12 (75%), and fundal rugae hypertrophies in 5 (80%). We suggest that the antral nodularity, raised erosions, mosaic appearance and mixed findings, are the reliable indicators of the underlying HP induced gastritis. However, these endoscopic findings are very specific, though not sensitive, for HP gastritis.