Detection of chromosome aberrations in the human interphase nucleus by visualization of specific target DNAs with radioactive and non-radioactive in situ hybridization techniques: diagnosis of trisomy 18 with probe L1.84.
The localization of chromosome 18 in human interphase nuclei is demonstrated by use of radioactive and non-radioactive in situ hybridization techniques with a DNA clone designated L1.84. This clone represents a distinct subpopulation of the repetitive human alphoid DNA family, located in the centric region of chromosome 18. Under stringent hybridization conditions hybridization of L1.84 is restricted to chromosome 18 and reflects the number of these chromosomes present in the nuclei, namely, two in normal diploid human cells and three in nuclei from cells with trisomy 18. Under conditions of low stringency, cross-hybridization with other subpopulations of the alphoid DNA family occurs in the centromeric regions of the whole chromosome complement, and numerous hybridization sites are detected over interphase nuclei. Detection of chromosome-specific target DNAs by non-radioactive in situ hybridization with appropriate DNA probes cloned from individual chromosomal subregions presents a rapid means of identifying directly numerical or even structural chromosome aberrations in the interphase nucleus. Present limitations and future applications of interphase cytogenetics are discussed.


Comparison of neoeriocitrin and naringin on proliferation and osteogenic differentiation in MC3T3-E1.
Naringin is considered the main effective compound of Drynaria Rhizome, which is used commonly in the treatment of osteoporosis in traditional Chinese medicine. However, we found neoeriocitrin, a new compound isolated from Drynaria Rhizome, showed a better activity than naringin on proliferation and osteogenic differentiation in MC3T3-E1. Both neoeriocitrin and naringin exhibited the best effect on proliferation and osteogenic differentiation at concentration of 2μg/ml. Neoeriocitrin more significantly improved proliferation and alkaline phosphatase (ALP) activity as well as up-regulated Runx2, COLI and OCN expression by 56%, 37% and 14% respectively than naringin. Furthermore, neoeriocitrin could rescue the inhibition effect of cell differentiation induced by PD98059 to some degree. Therefore, neoeriocitrin may be a new promising candidate drug for treatment of osteoporosis.


Phylogeny of Aging and Related Phenoptotic Phenomena.
The interpretation of aging as adaptive, i.e. as a phenomenon genetically determined and modulated, and with an evolutionary advantage, implies that aging, as any physiologic mechanism, must have phylogenetic connections with similar phenomena. This review tries to find the phylogenetic connections between vertebrate aging and some related phenomena in other species, especially within those phenomena defined as phenoptotic, i.e. involving the death of one or more individuals for the benefit of other individuals. In particular, the aim of the work is to highlight and analyze similarities and connections, in the mechanisms and in the evolutionary causes, between: (i) proapoptosis in prokaryotes and apoptosis in unicellular eukaryotes; (ii) apoptosis in unicellular and multicellular eukaryotes; (iii) aging in yeast and in vertebrates; and (iv) the critical importance of the DNA subtelomeric segment in unicellular and multicellular eukaryotes. In short, there is strong evidence that vertebrate aging has clear similarities and connections with phenomena present in organisms with simpler organization. These phylogenetic connections are a necessary element for the sustainability of the thesis of aging explained as an adaptive phenomenon, and, on the contrary, are incompatible with the opposite view of aging as being due to the accumulation of random damages of various kinds.


Local variations in reporting deaths to the coroner in England and Wales: a postcode lottery?
In England and Wales, doctors are charged with a responsibility either to report a death to the coroner or issue a medical certificate specifying cause of death. A lack of formal prescriptive or presumptive oversight has resulted in the promulgation by individual coroners of local reporting regimes. The study reported here identified overall and gendered variations in local reporting rates to coroners across the jurisdictions of England and Wales, consistent over time. Analysis was performed on Ministry of Justice (MOJ) data pertaining to the numbers and proportions of deaths reported to the coroner by jurisdiction over a 10-year period (2001-2010). Office of National Statistics (ONS) data provided the numbers of deaths registered in England and Wales over the same period to serve as a denominator for the calculation of proportions. Where coroner jurisdictions (and local authorities) had been amalgamated during this period, the combined reported and registered death figures have been included in line with the current jurisdiction areas. While reporting rates for individual jurisdictions were found to be stable over the 10-year period, wide local variations in reporting deaths to coroners were found with no obvious demographic explanation. The gender of the deceased was identified as a major factor in local variation. The decision to report a death to the coroner varies across jurisdictions. Implications for coronial investigations are discussed and the need for wider research into coroners' decision-making is proposed.


Testicular growth from birth to two years of age, and the effect of orchidopexy at age nine months: a randomized, controlled study.
To study whether surgical treatment at age 9 mo in boys with congenital unilaterally palpable undescended testes (cryptorchidism) is followed by improved growth of the previously retained testes compared to non-treatment. At the age of 6 mo, 70 boys were randomized to surgical treatment at 9 mo and 79 boys to treatment at 3 y of age. The boys were then followed at 12 and 24 mo. Ultrasonography was used to determine testicular volume. After orchidopexy, the previously retained testes resumed growth and were significantly larger than the non-operated testes at 2 y (0.49 ml vs 0.36 ml, p<0.001). Testicular growth after orchidopexy was also demonstrated by a higher mean ratio between the previously retained and the scrotal testes of the individual boys at 2 y: 0.84 for the surgically treated group, compared to 0.63 for the untreated group (p<0.001). Surgery at 9 mo has a beneficial effect on the growth of previously undescended testes.


Safety of task-shifting for male medical circumcision: a systematic review and meta-analysis.
Task-shifting for male medical circumcision is proposed as a strategy to overcome the lack of surgeons and doctors in high HIV prevalence settings. We undertook a systematic review and meta-analysis to review the safety of task-shifting for circumcision in Africa. We searched online databases and conference websites up to July 2011 without language restriction for studies reporting outcomes of task-shifting for circumcision in Africa. Information was extracted independently and in duplicate on study characteristics, quality, and outcome data. Case reports and case series were excluded. Ten studies met our inclusion criteria, providing outcome data on 25119 circumcisions. The proportion of adverse events ranged from 0.70 [95% confidence interval (CI) 0.44-1.02%] to 37.36% (95% CI 27.54-47.72%), with an overall pooled proportion of 2.31% (95% CI 1.46-3.16%; τ = 1.21; P < 0.001). Two studies reported outcomes separately for both doctors and non-physicians; there was no difference in the risk of adverse events by provider (pooled relative risk 1.18; 95% CI 0.78-1.78). The frequency of excessive bleeding ranged from 0.30 (0.09-0.65%) to 24.71% (16.27-34.26%) with an overall pooled prevalence of 0.55% (95% CI 0.13-0.97%). Infection occurred in 0.30 (0.14-1.47%) to 1.85% (0.07-5.96%) of cases, with an overall pooled proportion of 0.88% (95% CI 0.29-1.47%). All adverse events were reported to be non-severe. Task-shifting of male medical circumcision to non-physician clinicians can be done safely, with reported rates of adverse events similar to doctors and specialists.


Surveillance for antibiotic resistance in veterinary pathogens from the perspective of a regional diagnostic laboratory.
The Toowoomba Veterinary Laboratory tests for antibiotic resistance through passive surveillance of bacterial pathogens from diseased, frequently intensively managed, animals. Testing is carried out on the basis of the number of animals involved, the nature and severity of the disease and the identity and significance of the bacterium, the results guiding the submitting veterinarian in implementing appropriate treatment. The antibiotics chosen for testing are those that are currently registered for veterinary use and are considered effective in the given situation. Testing is carried out according to the current National Committee for Clinical Laboratory Standards Approved Standard for Disc Susceptibility Tests. This paper presents some results of testing bacterial pathogens from cattle and pigs.


Targeting inflammation to influence mood following spinal cord injury: a randomized clinical trial.
The purpose of the present study was to examine the efficacy of targeting inflammation as a means of improving mood following spinal cord injury (SCI) and explore the potential mechanisms of action. The study was a randomized, parallel-group, controlled, clinical trial (NCT02099890) whereby 20 participants with varying levels and severities of SCI were randomized (3:2) to either the treatment group, consisting of a 12-week anti-inflammatory diet, or control group. Outcome measures were assessed at baseline, 1 and 3 months, and consisted of CES-D scores of depression, markers of inflammation as assessed by various pro- and anti-inflammatory cytokines and several amino acids related to depression. A significant group × time interaction was found for CES-D (Center for Epidemiologic studies Depression Scale) score (p = 0.01), the TRP/LNAA (tryptophan/large neutral amino acid) ratio (p = 0.04), the composite score of pro-inflammatory cytokines (p = 0.04), IL-1β (interleukin-1 beta) (p = 0.04), and IFN-γ (interferon gamma) (p = 0.03). Pearson's r correlation showed significance between the ∆IL-1β and both the ∆CES-D score (r = 0.740, p < 0.01) and the ∆KYN/TRP (kynurenine/tryptophan) ratio (r = 0.536, p = 0.02). The ∆KYN/TRP ratio was also significantly correlated with the ∆CES-D score (r = 0.586, p = 0.01). Mediation analysis showed that the relationship between the ∆KYN/TRP ratio and the ∆CES-D score was mediated by the ∆IL-1β. Subgroup analysis showed that participants with high CES-D scores had significantly higher concentrations of IL-1β, and all correlations were maintained or strengthened within this subgroup. Overall, the results demonstrated the effectiveness of targeting inflammation as a means of improving mood in SCI, with potential mechanisms relating to the reduction in IL-1β and improvements in levels of neuroactive compounds related to the kynurenine pathway. Due to the limited sample size, results should be interpreted with caution; however, they are worthy of further examination due to the potential impact of inflammation on depression. ClinicalTrials.gov ID: NCT02099890 .


The crimson conundrum: heme toxicity and tolerance in GAS.
The massive erythrocyte lysis caused by the Group A Streptococcus (GAS) suggests that the β-hemolytic pathogen is likely to encounter free heme during the course of infection. In this study, we investigated GAS mechanisms for heme sensing and tolerance. We compared the minimal inhibitory concentration of heme among several isolates and established that excess heme is bacteriostatic and exposure to sub-lethal concentrations of heme resulted in noticeable damage to membrane lipids and proteins. Pre-exposure of the bacteria to 0.1 μM heme shortened the extended lag period that is otherwise observed when naive cells are inoculated into heme-containing medium, implying that GAS is able to adapt. The global response to heme exposure was determined using microarray analysis revealing a significant transcriptome shift that included 79 up regulated and 84 down regulated genes. Among other changes, the induction of stress-related chaperones and proteases, including groEL/ES (8x), the stress regulators spxA2 (5x) and ctsR (3x), as well as redox active enzymes were prominent. The heme stimulon also encompassed a number of regulatory proteins and two-component systems that are important for virulence. A three-gene cluster that is homologous to the pefRCD system of the Group B Streptococcus was also induced by heme. PefR, a MarR-like regulator, specifically binds heme with stoichiometry of 1:2 and protoporphyrin IX (PPIX) with stoichiometry of 1:1, implicating it is one of the GAS mediators to heme response. In summary, here we provide evidence that heme induces a broad stress response in GAS, and that its success as a pathogen relies on mechanisms for heme sensing, detoxification, and repair.


Effects of education to facilitate knowledge about chronic pain for adults: a systematic review with meta-analysis.
Chronic pain can contribute to disability, depression, anxiety, sleep disturbances, poor quality of life and increased health care costs, with close to 20 % of the adult population in Europe reporting chronic pain. To empower the person to self-manage, it is advocated that education and training about the nature of pain and its effects and how to live with pain is provided. The objective of this review is to determine the level of evidence for education to facilitate knowledge about chronic pain, delivered as a stand-alone intervention for adults, to reduce pain and disability. We identified randomised controlled trials of educational intervention for chronic pain by searching CENTRAL, MEDLINE, EMBASE and ongoing trials registries (inception to December 2013). Main inclusion criteria were (1) pain >3 months; (2) study design that allowed isolation of effects of education and (3) measures of pain or disability. Two reviewers independently screened and appraised each study. Nine studies were analysed. Pooled data from five studies, where the comparator group was usual care, showed no improvement in pain or disability. In the other four studies, comparing different types of education, there was no evidence for an improvement in pain; although, there was evidence (from one study) of a decrease in disability with a particular form of education-pain neurophysiology education (PNE). Post-hoc analysis of psychosocial outcomes reported in the studies showed evidence of a reduction in catastrophising and an increase of knowledge about pain following PNE. The evidence base is limited by the small numbers of studies, their relatively small sample sizes, and the diversity in types of education studied. From that limited evidence, the only support for this type of education is for PNE, though it is insufficiently strong to recommend conclusively that PNE should be delivered as a stand-alone intervention. It therefore remains sensible to recommend that education be delivered in conjunction with other pain management approaches as we cannot confidently conclude that education alone is effective in reducing pain intensity or related disability in chronic pain in adults.


Review article: the role of antibiotics vs. conventional pharmacotherapy in treating symptoms of irritable bowel syndrome.
The concept of augmenting the management of irritable bowel syndrome with antibiotics is evolving, and many questions remain regarding this therapy relative to known and hypothesized irritable bowel syndrome pathophysiology. The clinical evidence of small intestinal bacterial overgrowth as an important aetiology of irritable bowel syndrome continues to accumulate. Clinical symptoms of bacterial overgrowth and irritable bowel syndrome are similar; however, a definitive cause-and-effect relationship remains unproven. It is unclear whether motility dysfunction causes bacterial overgrowth or gas products of enteric bacteria affect intestinal motility in irritable bowel syndrome. To discusses the efficacy and tolerability of current symptom-directed pharmacotherapies and of antibiotics in the treatment of irritable bowel syndrome. A computerized search of PubMed was performed with search terms "IBS", "pharmacotherapy" and "antibiotics". Relevant articles were selected, and the reference list of selected articles was reviewed to identify additional references. Antibiotic treatment benefits a subset of irritable bowel syndrome patients. The non-absorbed antibiotic rifaximin has a favourable safety and tolerability profile compared with systemic antibiotics and demonstrates a therapeutic efficacy comparable with symptom-based irritable bowel syndrome pharmacotherapies. Rifaximin is the only antibiotic with demonstrated sustained benefit beyond therapy cessation in irritable bowel syndrome patients in a placebo-controlled trial. Whether antibiotics can improve quality of life in patients with irritable bowel syndrome warrants further research.


Effects of chronic lithium treatment on dopamine receptors in the rat corpus striatum. II. No effect on denervation or neuroleptic-induced supersensitivity.
The influence of chronic dietary lithium administration was evaluated on dopamine receptor supersensitivity in the rat corpus striatum. Supersensitivity was induced with either unilateral destruction of dopamine-containing fibers in the nigrostriatal pathway or with 3 weeks of treatment with haloperidol (HAL). Both treatments elevated [3H]spiroperidol binding sites, but in neither case was this increase in ligand binding affected by chronic dietary Li (brain levels 0.8 to 1.2 mEq/1 tissue). Our rats receiving 21 daily injections of HAL did show a behavioral supersensitivity to the dopamine agonist, apomorphine, and this effect was attenuated by concurrent treatment with dietary Li (accompanying paper). However, in contrast to previous data, this behavioral attenuation could not be linked to the prevention of increased [3H]spiroperidol binding in the corpus striatum. Furthermore, co-administration of dietary Li to subjects injected with HAL for 3 weeks did not reverse the increased density of [3H]spiroperidol binding sites which developed in the corpus striatum. Neither HAL nor Li treatment altered the affinity of the radioligand for its binding site. In the same animals, neostriatal dopamine-sensitive adenylate cyclase was not affected by either long-term dietary Li or chronic neuroleptic treatment, supporting the view that membrane antagonist and agonist sites differentially adapt to chronic alterations of synaptic input. Taken together, the results are incompatible with the hypothesis that the anti-manic action of Li is related to its ability to prevent dopamine receptor supersensitivity.


An empirical study using permutation-based resampling in meta-regression.
In meta-regression, as the number of trials in the analyses decreases, the risk of false positives or false negatives increases. This is partly due to the assumption of normality that may not hold in small samples. Creation of a distribution from the observed trials using permutation methods to calculate P values may allow for less spurious findings. Permutation has not been empirically tested in meta-regression. The objective of this study was to perform an empirical investigation to explore the differences in results for meta-analyses on a small number of trials using standard large sample approaches verses permutation-based methods for meta-regression. We isolated a sample of randomized controlled clinical trials (RCTs) for interventions that have a small number of trials (herbal medicine trials). Trials were then grouped by herbal species and condition and assessed for methodological quality using the Jadad scale, and data were extracted for each outcome. Finally, we performed meta-analyses on the primary outcome of each group of trials and meta-regression for methodological quality subgroups within each meta-analysis. We used large sample methods and permutation methods in our meta-regression modeling. We then compared final models and final P values between methods. We collected 110 trials across 5 intervention/outcome pairings and 5 to 10 trials per covariate. When applying large sample methods and permutation-based methods in our backwards stepwise regression the covariates in the final models were identical in all cases. The P values for the covariates in the final model were larger in 78% (7/9) of the cases for permutation and identical for 22% (2/9) of the cases. We present empirical evidence that permutation-based resampling may not change final models when using backwards stepwise regression, but may increase P values in meta-regression of multiple covariates for relatively small amount of trials.


Identifying causal gateways and mediators in complex spatio-temporal systems.
Identifying regions important for spreading and mediating perturbations is crucial to assess the susceptibilities of spatio-temporal complex systems such as the Earth's climate to volcanic eruptions, extreme events or geoengineering. Here a data-driven approach is introduced based on a dimension reduction, causal reconstruction, and novel network measures based on causal effect theory that go beyond standard complex network tools by distinguishing direct from indirect pathways. Applied to a data set of atmospheric dynamics, the method identifies several strongly uplifting regions acting as major gateways of perturbations spreading in the atmosphere. Additionally, the method provides a stricter statistical approach to pathways of atmospheric teleconnections, yielding insights into the Pacific-Indian Ocean interaction relevant for monsoonal dynamics. Also for neuroscience or power grids, the novel causal interaction perspective provides a complementary approach to simulations or experiments for understanding the functioning of complex spatio-temporal systems with potential applications in increasing their resilience to shocks or extreme events.


'Codeine is my companion': misuse and dependence on codeine containing medicines in Ireland.
Global concern around over the counter availability of codeine containing products and risk of misuse, dependence and related harms are evident. A phenomenological study of lived experiences of codeine misuse and dependence was undertaken in Ireland, following the Pharmaceutical Society of Ireland's 2010 guidelines for restricted supply of non-prescription codeine containing products. In-depth interviews were conducted with a purposive sample of adult codeine misusers and dependents (n=21), both actively using, in treatment and in recovery. The narratives were analysed using the Empirical Phenomenological Psychological five-step method (Karlsson, 1995). A total of 10 themes with 82 categories were identified. Two concepts at a higher level of abstraction above the theme-level emerged during the final stage of analysis. The concepts identified were 'emotional pain and user self-legitimization of use' and 'entrapment into habit-forming and invisible dependent use'. These concepts were reported in different ways by a majority of participants. Findings are presented under the following themes: (1) profile and product preferences; (2) awareness of habit forming use and harm; (3) negotiating pharmacy sales; (4) alternative sourcing routes; (5) the codeine feeling; (6) the daily routine; (7) acute and chronic side effects; (8) social isolation; (9) withdrawal and dependence and (10) help-seeking and treatment experiences. There is a public health and regulatory imperative to develop proactive responses tackling public availability of codeine containing medicines, risk minimisation in consumer self-treatment for pain, enhanced patient awareness of potential for habit forming use and its consequences and continued health professional pharmacovigilence.


Vocal fold wound healing after injection of human adipose-derived stem cells in a rabbit model.
Injection of injured rabbit vocal folds with human adipose-derived stem cells (hADSCs) led to improved wound healing and fewer signs of scarring as demonstrated by a decreased collagen content in the treated folds compared with the untreated folds. hADSCs remained viable for up to 12 weeks in rabbit vocal folds. The aim of this study was to investigate the morphologic and histologic properties of scarred rabbit vocal folds following injection of hADSCs. This was a randomized, controlled animal study. Twenty-four vocal folds from 12 New Zealand rabbits were scarred using a CO(2) laser and injected with either hADSCs (left vocal fold) or phosphate-buffered saline (right vocal fold). Every 4 weeks for the first 12 weeks after injection, an endoscopic examination was performed to assess the morphology of the vocal folds. Twelve weeks later the animals were euthanized and the tissues were stained for histology. In comparison with the right vocal folds, there was significantly less granulation tissue in the hADSCs-injected left vocal folds (p < 0.05). Histological examination revealed excessive collagen deposition and perichondral fibrosis in the right vocal folds, whereas the left vocal folds exhibited better wound healing and less collagen deposition (p < 0.05). Among the 12 specimens injected with hADSCs, 4 specimens demonstrated viable hADSCs under immunofluorescent cytochemistry.


Evolutionary insights into scleractinian corals using comparative genomic hybridizations.
Coral reefs belong to the most ecologically and economically important ecosystems on our planet. Yet, they are under steady decline worldwide due to rising sea surface temperatures, disease, and pollution. Understanding the molecular impact of these stressors on different coral species is imperative in order to predict how coral populations will respond to this continued disturbance. The use of molecular tools such as microarrays has provided deep insight into the molecular stress response of corals. Here, we have performed comparative genomic hybridizations (CGH) with different coral species to an Acropora palmata microarray platform containing 13,546 cDNA clones in order to identify potentially rapidly evolving genes and to determine the suitability of existing microarray platforms for use in gene expression studies (via heterologous hybridization). Our results showed that the current microarray platform for A. palmata is able to provide biological relevant information for a wide variety of coral species covering both the complex clade as well the robust clade. Analysis of the fraction of highly diverged genes showed a significantly higher amount of genes without annotation corroborating previous findings that point towards a higher rate of divergence for taxonomically restricted genes. Among the genes with annotation, we found many mitochondrial genes to be highly diverged in M. faveolata when compared to A. palmata, while the majority of nuclear encoded genes maintained an average divergence rate. The use of present microarray platforms for transcriptional analyses in different coral species will greatly enhance the understanding of the molecular basis of stress and health and highlight evolutionary differences between scleractinian coral species. On a genomic basis, we show that cDNA arrays can be used to identify patterns of divergence. Mitochondrion-encoded genes seem to have diverged faster than nuclear encoded genes in robust corals. Accordingly, this needs to be taken into account when using mitochondrial markers for scleractinian phylogenies.


Genetic variants in DNA repair genes and the risk of cutaneous malignant melanoma in melanoma-prone families with/without CDKN2A mutations.
Cutaneous malignant melanoma (CMM) is an etiologically heterogeneous disease with genetic, environmental (sun exposure) and host (pigmentation/nevi) factors and their interactions contributing to risk. Genetic variants in DNA repair genes may be particularly important since their altered function in response to sun exposure-related DNA damage maybe related to risk for CMM. However, systematic evaluations of genetic variants in DNA repair genes are limited, particularly in high-risk families. We comprehensively analyzed DNA repair gene polymorphisms and CMM risk in melanoma-prone families with/without CDKN2A mutations. A total of 586 individuals (183 CMM) from 53 families (23 CDKN2A (+), 30 CDKN2A (-)) were genotyped for 2964 tagSNPs in 131 DNA repair genes. Conditional logistic regression, conditioning on families, was used to estimate trend p-values, odds ratios and 95% confidence intervals for the association between CMM and each SNP separately, adjusted for age and sex. p-Values for SNPs in the same gene were combined to yield gene specific p-values. Two genes, POLN and PRKDC, were significantly associated with melanoma after Bonferroni correction for multiple testing (p = 0.0003 and 0.00035, respectively). DCLRE1B showed suggestive association (p = 0.0006). 28 ∼ 56% of genotyped SNPs in these genes had single SNP p < 0.05. The most significant SNPs in POLN and PRKDC had similar effects in CDKN2A (+) and CDKN2A (-) families. Our finding suggests that polymorphisms in DNA repair genes, POLN and PRKDC, were associated with increased melanoma risk in melanoma families with and without CDKN2A mutations.


A Simultaneous Analytical Method for Duplex Identification of Porcine and Horse in the Meat Products by EvaGreen based Real-time PCR.
A duplex real-time polymerase chain reaction (PCR) based assay for the detection of porcine and horse meat in sausages was designed by using EvaGreen fluorescent dye. Primers were selected from mitochondrial 12S rRNA and 16S rRNA genes which are powerful regions for identification of horse and porcine meat. DNA from reference samples and industrial products was successfully extracted using the GIDAGEN® Multi-Fast DNA Isolation Kit. Genomes were identified based on their specific melting peaks (Mp) which are 82.5℃ and 78℃ for horse and porcine, respectively. The assay used in this study allowed the detection of as little as 0.0001% level of horse meat and 0.001% level of porcine meat in the experimental admixtures. These findings indicate that EvaGreen based duplex real-time PCR is a potentially sensitive, reliable, rapid and accurate assay for the detection of meat species adulterated with porcine and horse meats.


Evidence-based chronic ulcer care and lower limb outcomes among Pacific Northwest veterans.
Evidence-based ulcer care guidelines detail optimal components of care for treatment of ulcers of different etiologies. We investigated the impact of providing specific evidence-based ulcer treatment components on healing outcomes for lower limb ulcers (LLU) among veterans in the Pacific Northwest. Components of evidence-based ulcer care for venous, arterial, diabetic foot ulcers/neuropathic ulcers were abstracted from medical records. The outcome was ulcer healing. Our analysis assessed the relationship between evidence-based ulcer care by etiology, components of care provided, and healing, while accounting for veteran characteristics. A minority of veterans in all three ulcer-etiology groups received the recommended components of evidence-based care in at least 80% of visits. The likelihood of healing improved when assessment for edema and infection were performed on at least 80% of visits (hazard ratio [HR] = 3.20, p = 0.009 and HR = 3.54, p = 0.006, respectively) in patients with venous ulcers. There was no significant association between frequency of care components provided and healing among patients with arterial ulcers. Among patients with diabetic/neuropathic ulcers, the chance of healing increased 2.5-fold when debridement was performed at 80% of visits (p = 0.03), and doubled when ischemia was assessed at the first visit (p = 0.045). Veterans in the Pacific Northwest did not uniformly receive evidence-based ulcer care. Not all evidence-based ulcer care components were significantly associated with healing. At a minimum, clinicians need to address components of ulcer care associated with improved ulcer healing.


Long-Term Economic and Employment Outcomes Among Partners of Women With Early-Stage Breast Cancer.
Work loss is one of many personal costs for patients with cancer and their families. Many women with breast cancer face long-term job loss that stems from their diagnoses. However, little is known about the economic and employment outcomes of partners of women with breast cancer. Women with nonmetastatic breast cancer identified by the Detroit and Los Angeles SEER registries between June 2005 and February 2007 were surveyed at both 9 months and 4 years after diagnosis. Partners were surveyed 4 years after patients' diagnoses. Nonretired partners were asked about employment and financial consequences that stemmed from the patients' breast cancer diagnoses and treatments. A total of 517 (67%) of 774 eligible partners completed the survey; 32% reported worsened financial status attributed to patients' breast cancers. Two hundred forty nonretired partners worked during the year after the patients' diagnoses; 90% were still working 4 years postdiagnosis. A total of 32% of partners decreased their work hours as a result of patients' breast cancer diagnoses and treatments; 64% of partners reported that, as a result of patients' breast cancer diagnoses and treatments, it was very/extremely important to keep working to keep health insurance. Overall, 51% of partners reported that it was very/extremely important to avoid changing jobs, because they were worried about loss of health insurance. Nearly one third of partners reported that their financial status was worse because of the patient's breast cancer, although most remained working 4 years after a diagnosis. Partners may continue to work longer than desired to compensate for a loss of financial resources in the family.


Suicide following self-harm: findings from the Multicentre Study of self-harm in England, 2000-2012.
Self-harm is a key risk factor for suicide and it is important to have contemporary information on the extent of risk. Mortality follow-up to 2012 of 40,346 self-harm patients identified in the three centres of the Multicentre Study of Self-harm in England between 2000 and 2010. Nineteen per cent of deaths during the study period (N=2704) were by suicide, which occurred in 1.6% of patients (2.6% of males and 0.9% of females), during which time the risk was 49 times greater than the general population risk. Overall, 0.5% of individuals died by suicide in the first year, including 0.82% of males and 0.27% of females. While the absolute risk of suicide was greater in males, the risk relative to that in the general population was higher in females. Risk of suicide increased with age. While self-poisoning had been the most frequent method of self-harm, hanging was the most common method of subsequent suicide, particularly in males. The number of suicides was probably a considerable underestimate as there were also a large number of deaths recorded as accidents, the majority of which were poisonings, these often involving psychotropic drugs. The study was focussed entirely on hospital-presenting self-harm. The findings underline the importance of prevention initiatives focused on the self-harm population, especially during the initial months following an episode of self-harm. Estimates using suicide and open verdicts may underestimate the true risk of suicide following self-harm; inclusion of accidental poisonings may be warranted in future risk estimates.


fMRI retinotopic mapping at 3 T: benefits gained from correcting the spatial distortions due to static field inhomogeneity.
fMRI retinotopic mapping usually relies upon Fourier analysis of functional responses to periodic visual stimuli that encode eccentricity or polar angle in the visual field. Generally, phase estimations are assigned to a surface model of the cerebral cortex and borders between retinotopic areas are eventually determined following ad hoc phase analysis on the surface model. Assigning functional responses to a surface model of the cortex is particularly sensitive to geometric distortions of the 3D functional data due to static field inhomogeneity. Here, we assess and document the benefits gained from correcting the fMRI data for these effects, under standard experimental conditions (echo-planar imaging, 3.0-T field strength) and with well-chosen acquisition parameters (regarding slice orientation and phase-encoding direction). While it appears that, in the absence of correction, errors in the estimates of the borders between low-order visual areas do not then significantly exceed the variance of statistical origin, about half of the functional responses in a retinotopic experiment are misassigned to neighboring functional areas. Therefore, correction of the effects due to geometric distortions is important in any retinotopic mapping experiment and by extension in any fMRI experiment on the visual system.


Immunophenotypic and immunoelectron microscopic characterization of major constituent cells in malignant fibrous histiocytoma using human cell lines and their transplanted tumors in immunodeficient mice.
Malignant fibrous histiocytoma (MFH) is the most common soft tissue sarcoma of adult life. Its histogenesis, however, is still a matter of debate because of its various cellular components. To elucidate the nature of tumor cells of MFH, six human MFH cell lines were compared immunohistochemically and immunoelectron microscopically with fibrosarcoma and fibroblast cell lines. In vitro differentiation of tumor cells was evaluated after the treatment with 12-O-tetradecanoylphorbol 13-acetate (TPA), IL-3, macrophage CSF, and granulocyte-macrophage CSF. Heterotransplantation of MFH tumor cells into nude mice and severe combined immunodeficient mice was performed to examine whether the tumor cells differentiate toward histiocytes or not. Gene expression of monocyte chemoattractant protein-1 in cultured tumor cells and transplanted tumors was also evaluated. All MFH cell lines examined were positive for collagen type I and prolyl hydroxylase. They failed to react with most anti-myeloid mAb such as CD11c, CD14, CD15, CD33, CD35, CD45, anti-HLA-DR, and PM-2K. Several Ab, including CD13, CD32, CD68, CD71, and HAM56, were reactive with MFH cells. However, all of these Ab were also reactive with fibrosarcoma and/or fibroblastic cell lines. These data indicate that MFH cell lines possess immunophenotypic characteristics very similar to those of fibrosarcoma and fibroblastic cell lines. In vitro treatment of tumor cells with TPA, IL-3, macrophage CSF, granulocyte-macrophage CSF, or their combination did not change their immunophenotypic characteristics and did not induce differentiation toward histiocytes (macrophages). Transplantation of tumor cells into nude mice and severe combined immunodeficient mice produced tumors similar in histology to human MFH. Tumor cells in the transplanted tumors revealed the same immunophenotypic characterization that they did in vitro. No in vivo differentiation of tumor cells toward histiocytes was observed. Immunohistochemical staining with anti-mouse macrophage mAb revealed marked infiltration of macrophages of mouse origin. Quantitative immunoelectron microscopy disclosed that these cells coincided with histiocyte-like cells. Gene expression of monocyte chemoattractant protein-1 was detected in all MFH cell lines as well as in transplanted tumors. The histiocyte-like cells in malignant fibrous histiocytoma are not a neoplastic component but rather infiltrated macrophages attracted by tumor-derived monocyte chemoattractant(s), and the tumor cells belong to a fibroblastic lineage differentiated from mesenchymal cells.


Lactic acidosis not hyperlactatemia as a predictor of in hospital mortality in septic emergency patients.
To compare the prognostic abilities of lactic acidosis and hyperlactatemia and determine the influence of metabolic acidosis when attempting to predict the outcome of septic patients using serum lactate levels. 126 patients with severe sepsis or septic shock were prospectively included in this study at an emergency department. Haemodynamic variables, simplified acute physiology score (SAPS) II, arterial blood gas studies and serum lactate levels were obtained at the time of presentation (0 h) and 4 h after presentation. The probability of mortality was calculated using SAPS II scores and compared with actual inhospital mortality at different serum lactate levels and arterial pH. Survival curves for lactic acidosis and severe hyperlactatemia were analysed using the Kaplan-Meier method. The calculated probability of mortality decreased from 35.7% (95% CI 30.2 to 41.2) at presentation to 29.3% (95% CI 24.2 to 34.4) at 4 h post-presentation. 27.0% of patients (34) died in hospital. Arterial pH and SAPS II scores were independent factors for predicting mortality of septic patients, at 0 h and 4 h. Serum lactate levels were closely related to severity of illness and metabolic acidosis in septic patients. Patients with lactic acidosis had significantly higher inhospital mortality than patients with normal pH and normal lactate levels by Kaplan-Meier survival analysis as determined based on measurements made at 0 h and 4 h (p<0.001, p<0.001 by the log-rank test, respectively). No significant difference in survival was found between patients with hyperlactatemia and those with normal pH and serum lactate levels. Lactic acidosis not hyperlactatemia was found to predict inhospital mortality more exactly in severe sepsis and septic shock patients. The acid-base state should be considered when attempting to predict the outcome of septic patients using serum lactate levels.


Lead effects on corn mitochondrial respiration.
Oxidation of exogenous nicotinamide-adenine dinucleotide and succinate by corn mitochondria was measured as a function of lead chloride concentration. Lead chloride (50 to 62 micromoles per liter) stimulated oxidation of exogenous reduced nicotinamide-adenine dinucleotide by 174 to 640 percent depending on the reaction mediums, whereas lead chloride (12.5 micromoles per liter) inhibited succinate oxidation by more than 80 percent. When inorganic phosphate was included in reaction mediums the subsequent addition of lead was without effect due to the low solubility of lead phosphate. If addition of lead was followed by addition of phosphate the inhibition of succinate oxidation by lead was released, but there was no reduction in the stimulation of oxidation of reduced nicotinamide-adenine dinucleotide by lead. The effects of lead on plant growth might be accentuated under conditions of phosphate deficiency.


Superfamilies SDR and MDR: from early ancestry to present forms. Emergence of three lines, a Zn-metalloenzyme, and distinct variabilities.
Two large gene and protein superfamilies, SDR and MDR (short- and medium-chain dehydrogenases/reductases), were originally defined from analysis of alcohol and polyol dehydrogenases. The superfamilies contain minimally 82 and 25 genes, respectively, in humans, minimally 324 and 86 enzyme families when known lines in other organisms are also included, and over 47,000 and 15,000 variants in existing sequence data bank entries. SDR enzymes have one-domain subunits without metal and MDR two-domain subunits without or with zinc, and these three lines appear to have emerged in that order from the universal cellular ancestor. This is compatible with their molecular architectures, present multiplicity, and overall distribution in the kingdoms of life, with SDR also of viral occurrence. An MDR-zinc, when present, is often, but not always, catalytic. It appears also to have a structural role in inter-domain interactions, coenzyme binding and substrate pocket formation, as supported by domain variability ratios and ligand positions. Differences among structural and catalytic zinc ions may be relative and involve several states. Combined, the comparisons trace evolutionary properties of huge superfamilies, with partially redundant enzymes in cellular redox functions.


Does it fit okay? Problems with condom use as a function of self-reported poor fit.
To identify associations between men's self-reports of ill-fitting condoms and selected condom use problems, using an event-specific analysis. A convenience sample of men was recruited via advertisements in newspapers (two urban and one small town) and a blog on the website of a condom sales company. Men completed a questionnaire posted on the website of The Kinsey Institute for Research in Sex, Gender, and Reproduction. Inclusion criteria were: at least 18 years old, used condoms for penile-vaginal intercourse in the past 3 months and the ability to read English. In controlled, event-specific, analyses of 436 men, those reporting ill-fitting condoms (44.7%) were significantly more likely to report breakage (adjusted odds ratio (AOR 2.6), slippage (AOR 2.7), difficulty reaching orgasm, both for their female partners (AOR 1.9) and for themselves (AOR 2.3). In addition, they were more likely to report irritation of the penis (AOR 5.0) and reduced sexual pleasure, both for their female partner (AOR 1.6) and for themselves (AOR 2.4). Furthermore, they were more likely to report that condoms interfered with erection (AOR 2.0), caused erection loss (AOR 2.3), or became dry during sex (AOR 1.9). Finally, they were more likely to report removing condoms before penile-vaginal sex ended (AOR 2.0). Men and their female sex partners may benefit from public health efforts designed to promote the improved fit of condoms.


Microglia-Secreted Galectin-3 Acts as a Toll-like Receptor 4 Ligand and Contributes to Microglial Activation.
Inflammatory response induced by microglia plays a critical role in the demise of neuronal populations in neuroinflammatory diseases. Although the role of toll-like receptor 4 (TLR4) in microglia's inflammatory response is fully acknowledged, little is known about endogenous ligands that trigger TLR4 activation. Here, we report that galectin-3 (Gal3) released by microglia acts as an endogenous paracrine TLR4 ligand. Gal3-TLR4 interaction was further confirmed in a murine neuroinflammatory model (intranigral lipopolysaccharide [LPS] injection) and in human stroke subjects. Depletion of Gal3 exerted neuroprotective and anti-inflammatory effects following global brain ischemia and in the neuroinflammatory LPS model. These results suggest that Gal3-dependent-TLR4 activation could contribute to sustained microglia activation, prolonging the inflammatory response in the brain.


Visual processing in Alzheimer's disease: surface detail and colour fail to aid object identification.
It has been suggested that object recognition in patients with Alzheimer's disease (AD) may be strongly influenced both by image format (e.g. colour vs. line-drawn) and by low-level visual impairments. To examine these notions, we tested basic visual functioning and picture naming in 41 AD patients and 40 healthy elderly controls. Picture naming was examined using 105 images representing a wide range of living and nonliving subcategories (from the Hatfield image test [HIT]: [Adlington, R. A., Laws, K. R., & Gale, T. M. (in press). The Hatfield image test (HIT): A new picture test and norms for experimental and clinical use. Journal of Clinical and Experimental Neuropsychology]), with each item presented in colour, greyscale, or line-drawn formats. Whilst naming for elderly controls improved linearly with the addition of surface detail and colour, AD patients showed no benefit from the addition of either surface information or colour. Additionally, controls showed a significant category by format interaction; however, the same profile did not emerge for AD patients. Finally, AD patients showed widespread and significant impairment on tasks of visual functioning, and low-level visual impairment was predictive of patient naming.


The result of treatment on vestibular and general pain thresholds in women with provoked vestibulodynia.
To correlate changes in vestibular pain thresholds to general pain thresholds in a subgroup of women with provoked vestibulodynia taking part in a treatment study. Thirty-five women with provoked vestibulodynia were randomized to 4 months' treatment with either electromyographic biofeedback (n=17) or topical lidocaine (n=18). Vestibular and general pressure pain thresholds (PPTs) were measured and the health survey Short Form-36 (SF-36) was filled out before treatment and at a 6-month follow-up. Subjective treatment outcome and bodily pain were analyzed. Thirty healthy women of the same age served as controls for general PPTs and SF-36. No differences in outcome measures were observed between the 2 treatments. Vestibular pain thresholds increased from median 30 g before to 70 g after treatment in the anterior vestibule (P<0.001) and from median 20 to 30 g in the posterior vestibule (P<0.001). PPTs on the leg and arm were lower in the patients as compared with controls both before and at the 6-month follow-up. Patients reporting total cure were 3/35; 25/35 were improved. The number of patients who frequently reported of other bodily pain was reduced after the treatment. The patients had lower scores for SF-36 (General Health, Vitality) before treatment, which was restored at the 6-month follow-up. Treating provoked vestibulodynia by either topical lidocaine or electromyographic biofeedback increased vestibular pain thresholds, reduced dyspareunia, and improved bodily pain. The patients showed a general hypersensitivity to pressure pain compared with controls and in this study the hypersensitivity did not seem to be affected by treating the superficial dyspareunia.


Standardization of Fluorescent Reporter Assays in Synthetic Biology across the Visible Light Spectrum.
In synthetic biology, Fluorescent reporters are frequently used to characterize the expression levels obtained from both genetic parts such as promoters and ribosome binding sites as well as from complex genetic circuits. To this end, plate readers offer an easy and high-throughput way of characterizing both the growth and fluorescence expression levels of cell cultures. However, despite the similar mode of action used in different devices, their output is not comparable due to intrinsic differences in their setup. Additionally, the generated output is expressed using arbitrary units, limiting reliable comparison of results to measurements taken within one single experiment using one specific plate reader, hampering the transferability of data across different plate readers and laboratories. This article presents an easy and accessible calibration method for transforming the device-specific output into a standardized output expressing the amount of fluorescence per well as a known equivalent fluorophore concentration per cell for fluorescent reporters spanning the visible light spectrum. This calibration method follows a 2-fold approach determining both the estimated number of cells and the equivalent chemical fluorophore concentration per well. It will contribute to the comparison of plate reader experiments between different laboratories across the world and will therefore greatly improve the reliability and exchange of both results and genetic parts between research groups.


Circle-to-circle amplification on a digital microfluidic chip for amplified single molecule detection.
We demonstrate a novel digital microfluidic nucleic acid amplification concept which is based on padlock probe mediated DNA detection and isothermal circle-to-circle amplification (C2CA). This assay platform combines two digital approaches. First, digital microfluidic manipulation of droplets which serve as micro-reaction chambers and shuttling magnetic particles between these droplets facilitates the integration of complex solid phase multistep assays. We demonstrate an optimized novel particle extraction and transfer protocol for superparamagnetic particles on a digital microfluidic chip that allows for nearly 100% extraction efficiencies securing high assay performance. Second, the compartmentalization required for digital single molecule detection is solved by simple molecular biological means, circumventing the need for complex microfabrication procedures necessary for most, if not all, other digital nucleic acid detection methods. For that purpose, padlock probes are circularized in a strictly target dependent ligation reaction and amplified through two rounds of rolling circle amplification, including an intermediate digestion step. The reaction results in hundreds of 500 nm sized individually countable DNA nanospheres per detected target molecule. We demonstrate that integrated miniaturized digital microfluidic C2CA results in equally high numbers of C2CA products μL(-1) as off-chip tube control experiments indicating high assay performance without signal loss. As low as 1 aM synthetic Pseudomonas aeruginosa DNA was detected with a linear dynamic range over 4 orders of magnitude up to 10 fM proving excellent suitability for infectious disease diagnostics.


Physical Activity, Global DNA Methylation, and Breast Cancer Risk: A Systematic Literature Review and Meta-analysis.
The extent to which physical activity reduces breast cancer risk through changes in global DNA methylation is unknown. We systematically identified studies that investigated the association between: (i) physical activity and global DNA methylation; or (ii) global DNA methylation and breast cancer risk. Associations were quantified using random-effects models. Heterogeneity was investigated through subgroup analyses and the Q-test and I 2 statistics. Twenty-four studies were reviewed. We observed a trend between higher levels of physical activity and higher levels of global DNA methylation [pooled standardized mean difference = 0.19; 95% confidence interval (CI), -0.03-0.40; P = 0.09] which, in turn, had a suggestive association with a reduced breast cancer risk (pooled relative risk = 0.70; 95% CI, 0.49-1.02; P = 0.06). In subgroup analyses, a positive association between physical activity and global DNA methylation was observed among studies assessing physical activity over long periods of time (P = 0.02). Similarly, the association between global DNA methylation and breast cancer was statistically significant for prospective cohort studies (P = 0.007). Despite the heterogeneous evidence base, the literature suggests that physical activity reduces the risk of breast cancer through increased global DNA methylation. This study is the first to systematically overview the complete biologic pathway between physical activity, global DNA methylation, and breast cancer. Cancer Epidemiol Biomarkers Prev; 27(11); 1320-31. ©2018 AACR.


Alpha adrenergic contributions to dysrhythmia during myocardial ischemia and reperfusion in cats.
Alpha compared to beta adrenergic contributions to dysrhythmias induced by left anterior descending coronary occlusion and by reperfusion were assessed in chloralose-anesthetized cats (n = 96). Alpha receptor blockade with either phentolamine or prazosin significantly reduced the number of premature ventricular complexes during coronary reperfusion (321 +/- 62-14 +/- 10 premature ventricular complexes, P less than 0.001), abolished early ventricular fibrillation (from 25% in controls to 0%), and prevented the increase in idioventricular rate seen with coronary reperfusion. However, beta-receptor blockade was without effect. Ventricular dysrhythmias induced by coronary occlusion alone (without reperfusion) were attenuated markedly by alpha-receptor blockade under conditions in which perfusion (measured with radiolabeled microspheres) within ischemic zones was not affected. Alternative sympatholytic interventions including pretreatment with 6-hydroxydopamine to deplete myocardial norepinephrine from 8.8 +/- 1.4 to 0.83 +/- 0.2 ng/mg protein and render the heart unresponsive to tyramine (120 microgram/kg) attenuated dysrhythmias induced by both coronary occlusion and reperfusion in a fashion identical to that seen with alpha-receptor blockade. Although efferent sympathetic activation induced by left stellate nerve stimulation increased idioventricular rate from 66 +/- 6 to 144+/- 7 beats/min (P less than 0.01) before coronary occlusion, this response was blocked by propranolol but not by phentolamine. In contrast, during reperfusion the increase in idioventricular rate induced by left stellate nerve stimulation (to 203 +/- 14) was not inhibited by propranolol but was abolished by phentolamine (79 +/- 10). Intracoronary methoxamine (0.1 microM) in animals depleted of myocardial catecholamines by 6-hydroxydopamine pretreatment did not affect idioventricular rate before coronary occlusion. However, early after coronary reperfusion, methoxamine increased idioventricular rate from 33 +/- 7 to 123 +/- 21 beats/min (P less than 0.01). Thus, enhanced alpha-adrenergic responsiveness occurs during myocardial ischemia and appears to be primary mediator of the electrophysiological derangements and resulting malignant dysrhythmias induced by catecholamines during myocardial ischemia and reperfusion.


SNPinfo: integrating GWAS and candidate gene information into functional SNP selection for genetic association studies.
We have developed a set of web-based SNP selection tools (freely available at http://www.niehs.nih.gov/snpinfo) where investigators can specify genes or linkage regions and select SNPs based on GWAS results, linkage disequilibrium (LD), and predicted functional characteristics of both coding and non-coding SNPs. The algorithm uses GWAS SNP P-value data and finds all SNPs in high LD with GWAS SNPs, so that selection is from a much larger set of SNPs than the GWAS itself. The program can also identify and choose tag SNPs for SNPs not in high LD with any GWAS SNP. We incorporate functional predictions of protein structure, gene regulation, splicing and miRNA binding, and consider whether the alternative alleles of a SNP are likely to have differential effects on function. Users can assign weights for different functional categories of SNPs to further tailor SNP selection. The program accounts for LD structure of different populations so that a GWAS study from one ethnic group can be used to choose SNPs for one or more other ethnic groups. Finally, we provide an example using prostate cancer and demonstrate that this algorithm can select a small panel of SNPs that include many of the recently validated prostate cancer SNPs.


Anencephalic infant with cleft palate and natal teeth: a case report.
Natal/neonatal teeth are very common in children with complete unilateral and bilateral cleft lip and palate. This article outlines a patient with intrauterine growth retardation, anencephaly, atrial septal defect, ventricular septal defect, two maxillary first natal incisor teeth, cleft palate, short neck, low-set ears, hypertelorism, retrognathia, and simian-line on the right hand. There is no conclusive evidence of a correlation between these findings and a known syndrome, suggesting that this case may be a hitherto undefined clinical combination with neonatal teeth.


The Swiss cohort of elderly patients with venous thromboembolism (SWITCO65+): rationale and methodology.
Venous thromboembolism (VTE) is common and has a high impact on morbidity, mortality, and costs of care. Although most of the patients with VTE are aged ≥65 years, there is little data about the medical outcomes in the elderly with VTE. The Swiss Cohort of Elderly Patients with VTE (SWITCO65+) is a prospective multicenter cohort study of in- and outpatients aged ≥65 years with acute VTE from all five Swiss university and four high-volume non-university hospitals. The goal is to examine which clinical and biological factors and processes of care drive short- and long-term medical outcomes, health-related quality of life, and medical resource utilization in elderly patients with acute VTE. The cohort also includes a large biobank with biological material from each participant. From September 2009 to March 2012, 1,863 elderly patients with VTE were screened and 1003 (53.8%) were enrolled in the cohort. Overall, 51.7% of patients were aged ≥75 years and 52.7% were men. By October 16, 2012, after an average follow-up time of 512 days, 799 (79.7%) patients were still actively participating. SWITCO65+ is a unique opportunity to study short- and long-term outcomes in elderly patients with VTE. The Steering Committee encourages national and international collaborative research projects related to SWITCO65+, including sharing anonymized data and biological samples.


Comparison of patterns of expression of tumour necrosis factor, lymphotoxin and interleukin-6 mRNA.
The expression of the mRNA encoding tumour necrosis factor, lymphotoxin and interleukin-6 by peripheral blood mononuclear cells was analysed. Unstimulated cells contained no detectable mRNA for these cytokines, however each mRNA was transiently expressed after stimulation with either the combination of phytohaemagglutinin and phorbol ester or the single stimulus of lipopolysaccharide. The dual stimulus yielded the stronger signal. The cytokine mRNA's had short half lives, but were stabilised following protein synthesis inhibition. Cyclosporin A completely blocked induction of lymphotoxin and partially inhibited induction of TNF and IL-6 mRNA. The features of regulation described in this paper suggest these genes belong within the "early" set of genes expressed following immune cell activation.


A mouse to human search for plasma proteome changes associated with pancreatic tumor development.
The complexity and heterogeneity of the human plasma proteome have presented significant challenges in the identification of protein changes associated with tumor development. Refined genetically engineered mouse (GEM) models of human cancer have been shown to faithfully recapitulate the molecular, biological, and clinical features of human disease. Here, we sought to exploit the merits of a well-characterized GEM model of pancreatic cancer to determine whether proteomics technologies allow identification of protein changes associated with tumor development and whether such changes are relevant to human pancreatic cancer. Plasma was sampled from mice at early and advanced stages of tumor development and from matched controls. Using a proteomic approach based on extensive protein fractionation, we confidently identified 1,442 proteins that were distributed across seven orders of magnitude of abundance in plasma. Analysis of proteins chosen on the basis of increased levels in plasma from tumor-bearing mice and corroborating protein or RNA expression in tissue documented concordance in the blood from 30 newly diagnosed patients with pancreatic cancer relative to 30 control specimens. A panel of five proteins selected on the basis of their increased level at an early stage of tumor development in the mouse was tested in a blinded study in 26 humans from the CARET (Carotene and Retinol Efficacy Trial) cohort. The panel discriminated pancreatic cancer cases from matched controls in blood specimens obtained between 7 and 13 mo prior to the development of symptoms and clinical diagnosis of pancreatic cancer. Our findings indicate that GEM models of cancer, in combination with in-depth proteomic analysis, provide a useful strategy to identify candidate markers applicable to human cancer with potential utility for early detection.


Mars: retention of ice.
Water in the form of ice can exist on Mars as permafrost that is either in equilibrium with the water content of the atmosphere or gradually evaporating through a protective layer of soil. The latter situation is evaluated quantitatively, and the required thicknesses of the protective layers are estimated. The presence of subsurface ice may explain the higher radar reflectivity of the dark areas than of the bright areas. Observation of its seasonal variations is suggested.


Effect of intracoronary thrombectomy on 30-day mortality in patients with acute myocardial infarction.
Insertion of intracoronary thrombectomy (ICT) devices, as a precedent to percutaneous coronary intervention (PCI), theoretically could have a beneficial effect on the outcome in patients with acute myocardial infarction. To examine whether ICT was associated with a lower 30-day mortality rate in patients with acute myocardial infarction, we studied 3,913 patients who underwent PCI within 24 hours after onset. A total of 990 patients (25.3%) were treated with ICT before PCI. The 30-day mortality rate was lower in the patients receiving ICT than in those without (3.7% vs 6.2%, p = 0.004), but this beneficial effect disappeared after adjustment for baseline characteristics (hazard ratio [HR] 0.658, p = 0.166). We also divided the patients into tertiles according to the Thrombolysis In Myocardial Infarction (TIMI) risk score. After adjustment for baseline characteristics, ICT was associated with a lower 30-day mortality rate in patients from the highest TIMI risk score tertile (HR 0.407, p = 0.029), but not in patients from the lower 2 tertiles. ICT was also an independent predictor of a lower 30-day mortality risk in patients aged > or =70 years (HR 0.239, p = 0.007), patients with diabetes mellitus (HR 0.275, p = 0.039), and those with stent implantation (HR 0.437, p = 0.034). In conclusion, in selected patients with high TIMI risk scores, an age > or =70 years, diabetes mellitus, or stenting, ICT is associated with a lower 30-day mortality rate.


Vision in elasmobranchs and their relatives: 21st century advances.
This review identifies a number of exciting new developments in the understanding of vision in cartilaginous fishes that have been made since the turn of the century. These include the results of studies on various aspects of the visual system including eye size, visual fields, eye design and the optical system, retinal topography and spatial resolving power, visual pigments, spectral sensitivity and the potential for colour vision. A number of these studies have covered a broad range of species, thereby providing valuable information on how the visual systems of these fishes are adapted to different environmental conditions. For example, oceanic and deep-sea sharks have the largest eyes amongst elasmobranchs and presumably rely more heavily on vision than coastal and benthic species, while interspecific variation in the ratio of rod and cone photoreceptors, the topographic distribution of the photoreceptors and retinal ganglion cells in the retina and the spatial resolving power of the eye all appear to be closely related to differences in habitat and lifestyle. Multiple, spectrally distinct cone photoreceptor visual pigments have been found in some batoid species, raising the possibility that at least some elasmobranchs are capable of seeing colour, and there is some evidence that multiple cone visual pigments may also be present in holocephalans. In contrast, sharks appear to have only one cone visual pigment. There is evidence that ontogenetic changes in the visual system, such as changes in the spectral transmission properties of the lens, lens shape, focal ratio, visual pigments and spatial resolving power, allow elasmobranchs to adapt to environmental changes imposed by habitat shifts and niche expansion. There are, however, many aspects of vision in these fishes that are not well understood, particularly in the holocephalans. Therefore, this review also serves to highlight and stimulate new research in areas that still require significant attention.


Antioxidant and Antisenescence Effects of Bergamot Juice.
Aging is one of the main risk factor for the onset of cardiovascular diseases; one of the possible explanations could be linked to the age-associated overproduction of free radicals. This increase of oxidative stress can be overcome with a high intake of food antioxidants. In this context, a number of studies have been addressed to assess the antiaging potential of natural antioxidant compounds. Recently, it has been shown that the juice of bergamot (Citrus bergamia Risso et Poiteau), a fruit mostly produced in the Ionian coastal areas of Southern Italy (Calabria), is a valuable source of health-promoting constituents with, among other, antioxidant properties. In order to investigate the potential antiaging effects of this Mediterranean natural antioxidant source, bergamot juices of three different cultivars ("fantastico," "femminello," and "castagnaro") were herein characterized by the mean of high-performance liquid chromatography-photodiode array-electrospray ionization-tandem mass spectrometry. Then, juices were investigated for the evaluation of total polyphenolic and flavonoid contents, cell-free model antioxidant activities, and in vitro antiaging properties on two different cellular models of induced myocardial senescence. The best performing juice was also assessed in vivo. The phytochemical profiles confirmed that juices were rich in flavonoids, both flavone and flavanone glycosides. In addition, two limonoid glycosides were also identified in all cultivars. Each cultivar showed different phenolic and flavonoid contents. In tube results showed the juice robust antioxidant activities that correlate with their phenolic and flavonoid contents. Moreover, for the first time, the ability of juice to counteract the chemical-induced senescence was here demonstrated in both cellular models. Lastly, the in vivo data obtained from mouse hearts evidenced an increase in transcription of genes involved in antiaging and antioxidant responses. The overall results suggest that bergamot juice exerts antioxidant and antisenescence effects, making it useful for nutraceutical purposes.


Rotavirus vaccination and intussusception - Science, surveillance, and safety: A review of evidence and recommendations for future research priorities in low and middle income countries.
As of January 2016, 80 countries have introduced rotavirus vaccines into their national immunization programs. Many have documented significant declines in rotavirus-specific and all-cause diarrheal illnesses following vaccine introduction. Two globally licensed rotavirus vaccines have been associated with a low risk of intussusception in several studies. In July 2014, the Rotavirus Organization of Technical Allies Council convened a meeting of research and advocacy organizations, public health experts, funders, and vaccine manufacturers to discuss post-marketing intussusception surveillance and rotavirus vaccine impact data. Meeting objectives were to evaluate updated data, identify and prioritize research gaps, discuss best practices for intussusception monitoring in lower-income settings and risk communication, and provide insight to country-level stakeholders on best practices for intussusception monitoring and communication. Meeting participants agreed with statements from expert bodies that the benefits of vaccination with currently available rotavirus vaccines outweigh the low risk of vaccination-associated intussusception. However, further research is needed to better understand the relationship of intussusception to wild-type rotavirus and rotavirus vaccines and delineate potential etiologies and mechanisms of intussusception. Additionally, evidence from research and post-licensure evaluations should be presented with evidence of the benefits of vaccination to best inform policymakers deciding on vaccine introduction or vaccination program sustainability.


Epigenetic modifications of the immune system in health and disease.
Vertebrate animals have developed sophisticated host defense mechanisms against potentially hostile antigens. These mechanisms mainly involve the immune system and the epithelial cells that cover the body surface. Accumulating studies have revealed that epigenetic mechanisms in collaboration with signal transduction networks regulate gene expression over the course of differentiation, proliferation and function of immune and epithelial cells. The epigenetic status of these cells is fine-tuned under physiological conditions; however, its disturbance often results in the development of immunological disorders, namely inflammation. Certain environmental factors influence the differentiation and function of immune cells through epigenetic alterations. For example, commensal microbiota-derived metabolites inhibit histone deacetylases to induce regulatory T cells, whereas some infectious agents induce DNA methylation, resulting in the development of cancer. These data imply that epigenetic regulation of host defense cells, which are usually the first to encounter external antigens, is implicated in disease development. Here, we highlight recent advances in our understanding of the molecular mechanisms by which the epigenetic status of immune and epithelial cells is controlled.


Hydroxyl radical production by H2O2-mediated oxidation of Fe(II) complexed by Suwannee River fulvic acid under circumneutral freshwater conditions.
The Fenton reaction, the oxidation of ferrous iron by hydrogen peroxide (H(2)O(2)), is typically assumed to be a source of hydroxyl radical (HO(•)) in natural systems, however, formation of HO(•) in this process is strongly dependent upon solution pH and the ligand environment, with HO(•) only formed when Fe(II) is organically complexed. In this study we examine the formation of HO(•) when Fe(II)-NOM complexes are oxidized by H(2)O(2) using phthalhydrazide as a probe for HO(•). We demonstrate that HO(•) formation can be quantitatively described using a kinetic model that assumes HO(•) formation occurs solely from the reaction of Fe(II)-NOM complexes with H(2)O(2), even though this reaction is sufficiently slow to play only a negligible role in the overall oxidation rate of total Fe(II). As such, NOM is seen to play a dual role in circumneutral natural systems in stabilizing Fe(II) toward oxidation by H(2)O(2) while enabling the formation of HO(•) through this oxidation process.


The -283C/T polymorphism of the DNMT3B gene influences the progression of joint destruction in rheumatoid arthritis.
The objective of this study is to investigate the association between the -283C/T polymorphism at the promotor of DNMT3B gene and susceptibility to rheumatoid arthritis (RA) and to evaluate the effect of the polymorphism on clinical features such as progression of joint destruction in RA. A total of 309 patients with RA were compared with 297 control subjects. Genotyping of the -283C/T polymorphism was performed by real-time sequencing using Pyrosequencer. The genotype frequencies of the polymorphism at position -283 were not significantly different between patients with RA and controls. There were significantly positive correlations between the modified Sharp score and the disease duration for carriers of each genotype (y = 9.546x + 19.998, p < 0.001, for T allele carriers, y = 6.185x + 34.424, p < 0.001 for CC homozygotes). The slope of regression line of the T allele carriers was significantly steeper than that of the CC homozygotes (p = 0.014). In conclusion, our results suggest that the -283C/T polymorphism of the DNMT3B gene is a genetic marker related to the joint destruction of RA.


Toward reliable characterization of energetic materials: interplay of theory and thermal analysis in the study of the thermal stability of tetranitroacetimidic acid (TNAA).
The thermal stability of energetic materials, being of the utmost importance for safety issues, is often considered in terms of kinetics, e.g., the Arrhenius parameters of the decomposition rate constant. The latter, in turn, are commonly determined using conventional thermoanalytical procedures with the use of simple Kissinger or Ozawa methods for kinetic data processing. However, thermal decomposition of energetic materials typically occurs via numerous exo- and endothermal processes including fast parallel reactions, phase transitions, autocatalysis, etc. This leads to numerous drawbacks of simple approaches. In this paper, we proposed a new methodology for characterization of the thermochemistry and thermal stability of melt-cast energetic materials, which is comprised of a complementary set of experimental and theoretical techniques in conjunction with a suitable kinetic model. With the aid of the proposed methodology, we studied in detail a novel green oxidizer, tetranitroacetimidic acid (TNAA). The experimental mass loss kinetics in the melt was perfectly fitted with a model comprised of zero-order reaction (sublimation or evaporation) and first-order thermal decomposition of TNAA with the effective Arrhenius parameters Ea = 41.0 ± 0.2 kcal mol-1 and log(A/s-1) = 20.2 ± 0.1. We rationalized the experimental findings on the basis of highly accurate CCSD(T)-F12 quantum chemical calculations. Computations predict that thermolysis of TNAA involves an intricate interplay of multiple decomposition channels of the three tautomers, which are equilibrated via either monomolecular reactions or concerted double hydrogen atom transfer in the H-bonded dimers; the calculated Arrhenius parameters of the effective rate constant coincide well with experiment. Most importantly, calculations provide detailed mechanistic evidence missing in the thermoanalytical experiment and explain formation of the experimentally observed primary products N2O and NO2. Along with the kinetics and mechanism of decomposition, the proposed approach yields accurate thermochemistry and phase change data of TNAA.


Ocular Morbidity in the Correction of Orbital Hypertelorism and Dystopia: A 15-Year Experience.
Complex surgery for appearance change is controversial. Correction of orbital hypertelorism risks diplopia and loss of stereopsis for aesthetic gain. The risk-to-benefit ratio remains ill-defined. The aim of this study was to define specific ocular morbidity following orbital translocation. The authors compared stable preoperative and postoperative orthoptic indices (i.e., angle of strabismus, ocular motility, and acuity) for 23 consecutive patients who underwent orbital translocation between 2000 and 2015 and noted the requirement for corrective surgery. Eighteen patients underwent 33 box osteotomies (15 bilateral and three unilateral). Five patients underwent facial bipartition. Diagnosis was craniofrontonasal dysplasia in 11, frontonasal dysplasia in six, facial cleft in four, and Saethre-Chotzen syndrome in two cases. Median grade of orbital hypertelorism was III. Median age at surgery was 13 years (range, 5 to 17 years). Nontransient change (favorable or unfavorable) in angle of strabismus was noted in 14 patients. Ocular motility was altered in 12. Six patients had stereopsis preoperatively that was disrupted in two cases because of changes in ocular alignment and/or motility. There was one perioperative ophthalmic emergency (superior orbital fissure syndrome) and three acute reexplorations for external ocular muscle entrapment. Corrective surgery for strabismus was performed for four patients. Three required secondary canthopexy and three required surgery for blepharoptosis. Orbital translocation is associated with ocular risks including changes to angle of strabismus, ocular motility, and (when present) loss of stereopsis. Secondary surgery to correct strabismus or eyelid malposition is common. Orbital translocation is an operative process, not a single procedure. Therapeutic, IV.


Social capital in an outdoor recreation context.
This study examined social capital development in three all-terrain vehicles (ATV) clubs in Maine using an adapted version of Lin's (2001) social capital theory model. The structural components of social capital identified included collective assets and individual assets in the form of normative behavior and trust relationships. Also identified were counter-norms for individual ATV riders identified as having divergent norms from club members. The second component of social capital is access to and mobilization of network contacts and resources. Access networks in the context of the ATV clubs studied were identified as community and landowner relations while mobilization of resources was existent in club membership attempts toward self-governance and efforts of the statewide "umbrella" organization. Instrumental outcomes benefit society and expressive outcomes benefit the individual. Both types of returns are present in the data suggesting that ATV clubs are creating social capital. This is important information to clubs who desire to market themselves, improve their reputations, and enhance their volunteer association. It is of further interest to state governments who fund clubs through trail grants as proof that a return on investment is being realized. Theoretical and applied implications for these and other types of recreation-based volunteer associations (e.g., clubs, friends groups, advocacy groups) are presented.


Heparanase activates the syndecan-syntenin-ALIX exosome pathway.
Exosomes are secreted vesicles of endosomal origin involved in signaling processes. We recently showed that the syndecan heparan sulfate proteoglycans control the biogenesis of exosomes through their interaction with syntenin-1 and the endosomal-sorting complex required for transport accessory component ALIX. Here we investigated the role of heparanase, the only mammalian enzyme able to cleave heparan sulfate internally, in the syndecan-syntenin-ALIX exosome biogenesis pathway. We show that heparanase stimulates the exosomal secretion of syntenin-1, syndecan and certain other exosomal cargo, such as CD63, in a concentration-dependent manner. In contrast, exosomal CD9, CD81 and flotillin-1 are not affected. Conversely, reduction of endogenous heparanase reduces the secretion of syntenin-1-containing exosomes. The ability of heparanase to stimulate exosome production depends on syntenin-1 and ALIX. Syndecans, but not glypicans, support exosome biogenesis in heparanase-exposed cells. Finally, heparanase stimulates intraluminal budding of syndecan and syntenin-1 in endosomes, depending on the syntenin-ALIX interaction. Taken together, our findings identify heparanase as a modulator of the syndecan-syntenin-ALIX pathway, fostering endosomal membrane budding and the biogenesis of exosomes by trimming the heparan sulfate chains on syndecans. In addition, our data suggest that this mechanism controls the selection of specific cargo to exosomes.


Atrial natriuretic factor. Possible implications in liver disease.
The discovery of the first well-defined natriuretic hormone, the Atrial Natriuretic Factor (ANF), has prompted research on its impact on volume regulation in health and disease. The natriuretic, diuretic, and smooth muscle-relaxing properties suggest an important role of this novel hormone in pathophysiological states with sodium or volume retention, such as congestive heart failure or cirrhosis of the liver. Investigations on the implications of ANF in liver disease have been performed for little more than 1 year, and results are still controversial in many respects. At present, it seems very likely that there is no absolute deficiency of plasma ANF in patients with cirrhosis. Moreover, elevated plasma levels in cirrhotics with ascites have been reported by several groups. However, as yet, a molecular characterization of this increased immunoreactivity is still lacking. There is disagreement on the reduced release of and renal response to ANF in subgroups of cirrhotics; however, stimulus-response-coupling might be impaired. Further studies are needed to elucidate the pathophysiological implications and therapeutical potential of ANF in patients with chronic liver disease.


Neutrophils promote alveolar epithelial regeneration by enhancing type II pneumocyte proliferation in a model of acid-induced acute lung injury.
Alveolar epithelial regeneration is essential for resolution of the acute respiratory distress syndrome (ARDS). Although neutrophils have traditionally been considered mediators of epithelial damage, recent studies suggest they promote type II pneumocyte (AT2) proliferation, which is essential for regenerating alveolar epithelium. These studies did not, however, evaluate this relationship in an in vivo model of alveolar epithelial repair following injury. To determine whether neutrophils influence alveolar epithelial repair in vivo, we developed a unilateral acid injury model that creates a severe yet survivable injury with features similar to ARDS. Mice that received injections of the neutrophil-depleting Ly6G antibody had impaired AT2 proliferation 24 and 72 h after acid instillation, which was associated with decreased reepithelialization and increased alveolar protein concentration 72 h after injury. As neutrophil depletion itself may alter the cytokine response, we questioned the contribution of neutrophils to alveolar epithelial repair in neutropenic granulocyte-colony stimulating factor (G-CSF)-/- mice. We found that the loss of G-CSF recapitulated the neutrophil response of Ly6G-treated mice and was associated with defective alveolar epithelial repair, similar to neutrophil-depleted mice, and was reversed by administration of exogenous G-CSF. To approach the mechanisms, we employed an unbiased protein analysis of bronchoalveolar lavage fluid from neutrophil-depleted and neutrophil-replete mice 12 h after inducing lung injury. Pathway analysis identified significant differences in multiple signaling pathways that may explain the differences in epithelial repair. These data emphasize an important link between the innate immune response and tissue repair in which neutrophils promote alveolar epithelial regeneration.


Minimal Change in the cytoplasmic calcium dynamics in striatal GABAergic neurons of a DYT1 dystonia knock-in mouse model.
DYT1 dystonia is the most common hereditary form of primary torsion dystonia. This autosomal-dominant disorder is characterized by involuntary muscle contractions that cause sustained twisting and repetitive movements. It is caused by an in-frame deletion in the TOR1A gene, leading to the deletion of a glutamic acid residue in the torsinA protein. Heterozygous knock-in mice, which reproduce the genetic mutation in human patients, have abnormalities in synaptic transmission at the principal GABAergic neurons in the striatum, a brain structure that is involved in the execution and modulation of motor activity. However, whether this mutation affects the excitability of striatal GABAergic neurons has not been investigated in this animal model. Here, we examined the excitability of cultured striatal neurons obtained from heterozygous knock-in mice, using calcium imaging as indirect readout. Immunofluorescence revealed that more than 97% of these neurons are positive for a marker of GABAergic neurons, and that more than 92% are also positive for a marker of medium spiny neurons, indicating that these are mixed cultures of mostly medium spiny neurons and a few (~5%) GABAergic interneurons. When these neurons were depolarized by field stimulation, the calcium concentration in the dendrites increased rapidly and then decayed slowly. The amplitudes of calcium transients were larger in heterozygous neurons than in wild-type neurons, resulting in ~15% increase in cumulative calcium transients during a train of stimuli. However, there was no change in other parameters of calcium dynamics. Given that calcium dynamics reflect neuronal excitability, these results suggest that the mutation only slightly increases the excitability of striatal GABAergic neurons in DYT1 dystonia.


Microarray gene expression data with linked survival phenotypes: diffuse large-B-cell lymphoma revisited.
Diffuse large-B-cell lymphoma (DLBCL) is an aggressive malignancy of mature B lymphocytes and is the most common type of lymphoma in adults. While treatment advances have been substantial in what was formerly a fatal disease, less than 50% of patients achieve lasting remission. In an effort to predict treatment success and explain disease heterogeneity clinical features have been employed for prognostic purposes, but have yielded only modest predictive performance. This has spawned a series of high-profile microarray-based gene expression studies of DLBCL, in the hope that molecular-level information could be used to refine prognosis. The intent of this paper is to reevaluate these microarray-based prognostic assessments, and extend the statistical methodology that has been used in this context. Methodological challenges arise in using patients' gene expression profiles to predict survival endpoints on account of the large number of genes and their complex interdependence. We initially focus on the Lymphochip data and analysis of Rosenwald et al. (2002). After describing relationships between the analyses performed and gene harvesting (Hastie et al., 2001a), we argue for the utility of penalized approaches, in particular least angle regression-least absolute shrinkage and selection operator (Efron et al., 2004). While these techniques have been extended to the proportional hazards/partial likelihood framework, the resultant algorithms are computationally burdensome. We develop residual-based approximations that eliminate this burden yet perform similarly. Comparisons of predictive accuracy across both methods and studies are effected using time-dependent receiver operating characteristic curves. These indicate that gene expression data, in turn, only delivers modest predictions of posttherapy DLBCL survival. We conclude by outlining possibilities for further work.


The effects of trust in authority and procedural fairness on cooperation.
The present research examined the effect of procedural fairness and trust in an authority on people's willingness to cooperate with the authority across a wide range of social situations. Prior research has shown that the presence of information about whether an authority can be trusted moderates the effect of procedural fairness. If no trust information is available, procedural fairness influences people's reactions. This is not the case when information about the trustworthiness of the authority is present. In the present article, it is argued that information about whether the authority can or cannot be trusted may also moderate the effect of procedural fairness in predicting levels of cooperation. Assuming that the use of fair procedures by authorities that cannot be trusted is less influential than is the enactment of procedures by trustworthy authorities, it is predicted that trust in authority moderates the influence of procedural fairness on cooperation in such a way that procedural fairness has a positive effect on cooperation primarily when trust in authority is high. Results from 4 studies (2 experimental studies and 2 field studies) provide supportive evidence for this interaction.


The Pandolf equation under-predicts the metabolic rate of contemporary military load carriage.
This investigation assessed the accuracy of error of the Pandolf load carriage energy expenditure equation when simulating contemporary military conditions (load distribution, external load and walking speed). Within-participant design. Sixteen male participants completed 10 trials comprised of five walking speeds (2.5, 3.5, 4.5, 5.5 and 6.5km·h-1) and two external loads (22.7 and 38.4kg). The Pandolf equation demonstrated poor predictive precision, with a mean bias of 124.9W and -48.7 to 298.5W 95% limits of agreement. Furthermore, the Pandolf equation systematically under-predicted metabolic rate (p<0.05) across the 10 speed-load combinations. Predicted metabolic rate error ranged from 12-33% across all conditions with the 'moderate' walking speeds (i.e. 4.5-5.5km·h-1) yielding less prediction error (12-17%) when compared to the slower and faster walking speeds (21-33%). Factors such as mechanical efficiency and load distribution contribute to the impaired predictive accuracy. The authors suggest the Pandolf equation should be applied to military load carriage with caution.


Emotional reactivity and parenting sensitivity interact to predict cortisol output in toddlers.
Cortisol output in response to emotion induction procedures was examined at child age 24 months in a prospective longitudinal sample of 1,292 children and families in predominantly low-income and nonurban communities in two regions of high poverty in the United States. Multilevel analysis indicated that observed emotional reactivity to a mask presentation but not a toy removal procedure interacted with sensitive parenting to predict cortisol levels in children. For children experiencing high levels of sensitive parenting, cortisol output was high among children exhibiting high emotional reactivity and low among children exhibiting low emotional reactivity. For children experiencing low levels of sensitive parenting, cortisol output was unrelated to emotional reactivity.


Minitags for small molecules: detecting targets of reactive small molecules in living plant tissues using 'click chemistry'.
Small molecules offer unprecedented opportunities for plant research since plants respond to, metabolize, and react with a diverse range of endogenous and exogenous small molecules. Many of these small molecules become covalently attached to proteins. To display these small molecule targets in plants, we introduce a two-step labelling method for minitagged small molecules. Minitags are small chemical moieties (azide or alkyne) that are inert under biological conditions and have little influence on the membrane permeability and specificity of the small molecule. After labelling, proteomes are extracted under denaturing conditions and minitagged proteins are coupled to reporter tags through a 'click chemistry' reaction. We introduce this two-step labelling procedure in plants by studying the well-characterized targets of E-64, a small molecule cysteine protease inhibitor. In contrast to biotinylated E-64, minitagged E-64 efficiently labels vacuolar proteases in vivo. We displayed, purified and identified targets of a minitagged inhibitor that targets the proteasome and cysteine proteases in living plant cells. Chemical interference assays with inhibitors showed that MG132, a frequently used proteasome inhibitor, preferentially inhibits cysteine proteases in vivo. The two-step labelling procedure can be applied on detached leaves, cell cultures, seedlings and other living plant tissues and, when combined with photoreactive groups, can be used to identify targets of herbicides, phytohormones and reactive small molecules selected from chemical genetic screens.


Synthesis and transdermal permeation of novel N4-methoxypoly(ethylene glycol) carbamates of cytarabine.
Cytarabine is a deoxycytidine analogue commonly used in the treatment of hematological malignant diseases. Its clinical utility, however, is severely limited by its short plasma half-life because of the catabolic action of nucleoside deaminases. In this study, N(4)-carbamate derivatives of cytarabine (1) were synthesized and evaluated for transdermal penetration because this mode of administration may circumvent its limitations. The synthesis of these compounds was achieved in a two-step process. First, the methoxypoly(ethylene glycol) was activated by p-nitrophenyl chloroformate. Second, the activated intermediates were reacted with cytarabine in the presence of N-hydroxysuccinamide to give the N(4)-methoxypoly(ethylene glycol) carbamate derivatives. The transdermal flux values of the N(4)-carbamates of cytarabine were determined in vitro by Franz diffusion cell methodology. Aqueous solubility and log D (pH 7.4) values were determined and assessed for correlation with transdermal flux values. The synthesized carbamates, particularly, (9)-(13), showed increased solubility in both aqueous and lipid media. Log D values decreased as the oxyethylene chain lengthened. Although none of the derivatives showed significantly higher transdermal penetration than cytarabine (1), it should be mentioned that the mean for cytarabine N(4)-methoxyethyleneoxycarbamate (8) was 10 times higher and the median was 2 times higher.


Perturbed connectivity of the amygdala and its subregions with the central executive and default mode networks in chronic pain.
Maladaptive responses to pain-related distress, such as pain catastrophizing, amplify the impairments associated with chronic pain. Many of these aspects of chronic pain are similar to affective distress in clinical anxiety disorders. In light of the role of the amygdala in pain and affective distress, disruption of amygdalar functional connectivity in anxiety states, and its implication in the response to noxious stimuli, we investigated amygdala functional connectivity in 17 patients with chronic low back pain and 17 healthy comparison subjects, with respect to normal targets of amygdala subregions (basolateral vs centromedial nuclei), and connectivity to large-scale cognitive-emotional networks, including the default mode network, central executive network, and salience network. We found that patients with chronic pain had exaggerated and abnormal amygdala connectivity with central executive network, which was most exaggerated in patients with the greatest pain catastrophizing. We also found that the normally basolateral-predominant amygdala connectivity to the default mode network was blunted in patients with chronic pain. Our results therefore highlight the importance of the amygdala and its network-level interaction with large-scale cognitive/affective cortical networks in chronic pain, and help link the neurobiological mechanisms of cognitive theories for pain with other clinical states of affective distress.


The effect of different suture removal time intervals on surgical wound healing.
This study was carried out to compare the effect of different suture removal time on surgical wound healing. Twenty-one male albino rabbits were used. Under general and local anesthesia a moucoperiosteal rectangular flap was raised in each animal and then repositioned and sutured. The animals were randomly divided into three experimental groups of seven animals each. In group I and II the sutures were removed after 3 and 5 days respectively and were followed up for 7 and 14 days after surgery. In group III the sutures were removed after 7 days and were followed up for 14 days after surgery. Tissue reactions were observed and recorded using inflammation and gingival indexes at 7 and 14 days after surgery in all three groups. Inflammation and gingival indexes were analyzed by Kurskal Wallis, Firedman and Wilcoxone tests. Results showed that inflammation index was significantly different with two other groups at the day 7 after surgery (P<0.008). Gingival index in group II was significantly different from two other groups at the day 14 (P<0.028); however, there was no significant difference between group II and III at the same interval. Based on result of this study, 5 days was recognized to the best time interval for suture removal in comparison with two other time intervals.