Comprehensive functional annotation of 77 prostate cancer risk loci.
Genome-wide association studies (GWAS) have revolutionized the field of cancer genetics, but the causal links between increased genetic risk and onset/progression of disease processes remain to be identified. Here we report the first step in such an endeavor for prostate cancer. We provide a comprehensive annotation of the 77 known risk loci, based upon highly correlated variants in biologically relevant chromatin annotations--we identified 727 such potentially functional SNPs. We also provide a detailed account of possible protein disruption, microRNA target sequence disruption and regulatory response element disruption of all correlated SNPs at r(2) ≥ 0.88%. 88% of the 727 SNPs fall within putative enhancers, and many alter critical residues in the response elements of transcription factors known to be involved in prostate biology. We define as risk enhancers those regions with enhancer chromatin biofeatures in prostate-derived cell lines with prostate-cancer correlated SNPs. To aid the identification of these enhancers, we performed genomewide ChIP-seq for H3K27-acetylation, a mark of actively engaged enhancers, as well as the transcription factor TCF7L2. We analyzed in depth three variants in risk enhancers, two of which show significantly altered androgen sensitivity in LNCaP cells. This includes rs4907792, that is in linkage disequilibrium (r(2) = 0.91) with an eQTL for NUDT11 (on the X chromosome) in prostate tissue, and rs10486567, the index SNP in intron 3 of the JAZF1 gene on chromosome 7. Rs4907792 is within a critical residue of a strong consensus androgen response element that is interrupted in the protective allele, resulting in a 56% decrease in its androgen sensitivity, whereas rs10486567 affects both NKX3-1 and FOXA-AR motifs where the risk allele results in a 39% increase in basal activity and a 28% fold-increase in androgen stimulated enhancer activity. Identification of such enhancer variants and their potential target genes represents a preliminary step in connecting risk to disease process.


Upregulation of KSRP by miR-27b provides IFN-γ-induced post-transcriptional regulation of CX3CL1 in liver epithelial cells.
Aberrant cellular responses to pro-inflammatory cytokines, such as IFN-γ, are pathogenic features in many chronic inflammatory diseases. A variety of feedback regulatory pathways have evolved to prevent an inappropriate cellular reaction to these pro-inflammatory cytokines. CX3CL1 is a unique chemokine and plays an important role in chronic liver diseases. We report here that IFN-γ stimulation induces a transient CX3CL1 production in liver epithelial cells (i.e., hepatocytes and biliary epithelial cells). This transient CX3CL1 production is accompanied with a destabilization of CX3CL1 mRNA associated with the induction of the KH-type splicing regulatory protein (KSRP). IFN-γ treatment of liver epithelial cells decreases expression level of miR-27b, a miRNA that targets the 3' untranslated region of KSRP mRNA resulting in translational suppression. Induction of KSRP following IFN-γ stimulation depends on the downregulation of miR-27b. Functional manipulation of KSRP or miR-27b caused reciprocal alterations in CX3CL1 mRNA stability in liver epithelial cells. Moreover, transfection of miR-27b precursor influences CX3CL1-associated chemotaxis effects of biliary epithelial cells to Jurkat T cells. These findings suggest that miR-27b-mediated post-transcriptional suppression controls the expression of KSRP in liver epithelial cells, and upregulation of KSRP destabilizes CX3CL1 mRNA, providing fine-tuning of cellular inflammatory reactions in response to IFN-γ stimulation.


The dynamic three-dimensional organization of the diploid yeast genome.
The budding yeast Saccharomyces cerevisiae is a long-standing model for the three-dimensional organization of eukaryotic genomes. However, even in this well-studied model, it is unclear how homolog pairing in diploids or environmental conditions influence overall genome organization. Here, we performed high-throughput chromosome conformation capture on diverged Saccharomyces hybrid diploids to obtain the first global view of chromosome conformation in diploid yeasts. After controlling for the Rabl-like orientation using a polymer model, we observe significant homolog proximity that increases in saturated culture conditions. Surprisingly, we observe a localized increase in homologous interactions between the HAS1-TDA1 alleles specifically under galactose induction and saturated growth. This pairing is accompanied by relocalization to the nuclear periphery and requires Nup2, suggesting a role for nuclear pore complexes. Together, these results reveal that the diploid yeast genome has a dynamic and complex 3D organization.


Child Nutrition Programs: Flexibilities for Milk, Whole Grains, and Sodium Requirements. Interim final rule.
This interim final rule extends through school year 2018-2019 three menu planning flexibilities currently available to many Child Nutrition Program operators, giving them near-term certainty about Program requirements and more local control to serve nutritious and appealing meals to millions of children nationwide. These flexibilities include: Providing operators the option to offer flavored, low-fat (1 percent fat) milk in the Child Nutrition Programs; extending the State agencies' option to allow individual school food authorities to include grains that are not whole grain-rich in the weekly menu offered under the National School Lunch Program (NSLP) and School Breakfast Program (SBP); and retaining Sodium Target 1 in the NSLP and SBP. This interim final rule addresses significant challenges faced by local operators regarding milk, whole grains and sodium requirements and their impact on food development and reformulation, menu planning, and school food service procurement and contract decisions. The comments from the public on the long-term availability of these three flexibilities will help inform the development of a final rule, which is expected to be published in fall 2018 and implemented in school year 2019-2020.


Adaptation of lymphadenopathy associated virus (LAV) to replication in EBV-transformed B lymphoblastoid cell lines.
A strain of lymphadenopathy associated retrovirus ( LAV ) passaged in vitro was used to infect a lymphoblastoid cell line obtained by transformation with Epstein-Barr virus of B lymphocytes from a healthy donor. The virus produced from this line (B- LAV ) was also able to grow at a high rate in some other lymphoblastoid lines and in a Burkitt lymphoma line. This adapted strain retained the biochemical, ultrastructural, and antigenic characteristics of the original strain, as well as its tropism for normal T4+ lymphocytes. It is thus possible to grow LAV in large quantities that can be used for the preparation of diagnostic reagents. The interaction between such a human retrovirus and Epstein-Barr virus, a DNA virus, may have some implication for the pathology of the acquired immunodeficiency syndrome and related diseases.


Carbon Nanomaterials Interfacing with Neurons: An In vivo Perspective.
Developing new tools that outperform current state of the art technologies for imaging, drug delivery or electrical sensing in neuronal tissues is one of the great challenges in neurosciences. Investigations into the potential use of carbon nanomaterials for such applications started about two decades ago. Since then, numerous in vitro studies have examined interactions between these nanomaterials and neurons, either by evaluating their compatibility, as vectors for drug delivery, or for their potential use in electric activity sensing and manipulation. The results obtained indicate that carbon nanomaterials may be suitable for medical therapies. However, a relatively small number of in vivo studies have been carried out to date. In order to facilitate the transformation of carbon nanomaterial into practical neurobiomedical applications, it is essential to identify and highlight in the existing literature the strengths and weakness that different carbon nanomaterials have displayed when probed in vivo. Unfortunately the current literature is sometimes sparse and confusing. To offer a clearer picture of the in vivo studies on carbon nanomaterials in the central nervous system, we provide a systematic and critical review. Hereby we identify properties and behavior of carbon nanomaterials in vivo inside the neural tissues, and we examine key achievements and potentially problematic toxicological issues.


Maimonides: an early but accurate view on the treatment of haemorrhoids.
Moses Maimonides was not only one of the most influential religious figures of the middle ages, but also a pioneer in a wide variety of medical practices. A brief history of his life, and what is known about his medical education, is given here. His paper on haemorrhoids is summarised, as well as a review of the current understanding of the pathogenesis, prevention and treatment of this common condition. The comparison of Maimonides' writings to modern understanding of not only the prevention and treatment of haemorrhoids, but also his approach to the patient as a whole in terms of pre- and postoperative care, demonstrate how ahead of his time this great philosopher was.


Anisotropy of light propagation in human skin.
Using spatially resolved, steady state diffuse reflectometry, a directional dependence was found in the propagation of visible and near infrared light through human skin in vivo. The skin's reduced scattering coefficient mu(s)' varies by up to a factor of two between different directions of propagation at the same position. This anisotropy is believed to be caused by the preferential orientation of collagen fibres in the dermis, as described by Langer's skin tension lines. Monte Carlo simulations that examine the effect of partial collagen fibre orientation support this hypothesis. The observation has consequences for non-invasive diagnostic methods relying on skin optical properties, and it could be used non-invasively to determine the direction of lines of cleavage in order to minimize scars due to surgical incisions.


Diabetic control and the renin-angiotensin system, catecholamines, and blood pressure.
Diabetic ketoacidosis is usually associated with marked secondary hyperaldosteronism. Plasma levels of renin, angiotensin II, and aldosterone are markedly raised before treatment in most patients, with values falling rapidly toward normal as metabolic control is restored. In a few patients, mostly those with long-term complications of diabetes, plasma levels of renin, angiotensin II, and aldosterone before treatment remain within the normal range. In moderately hyperglycemic patients who have glycosuria but not ketonuria, plasma levels of all three substances are significantly higher than when control is improved. Occasionally, moderately hyperglycemic patients have mild secondary hyperaldosteronism. Improved metabolic control in such patients causes a rise in plasma volume and a rise in total exchangeable sodium, the latter to levels significantly above normal. Plasma catecholamine levels are markedly elevated in diabetic ketoacidosis, probably as a consequence of the ketoacidotic state. In nonketotic patients with moderate hyperglycemia, basal plasma norepinephrine levels are normal; catecholamine responses to exercise may be exaggerated, however. Epidemiological and animal studies suggest a relationship between blood pressure and blood glucose levels. There are few clinical studies of the effects of altering metabolic control of diabetes on blood pressure, and this is an important area for further study.


Current legal responses to elder abuse.
Abstract. In this paper, legal provisions for dealing with elder abuse in social or healthcare settings will be considered. The need for such measures for detection and management of elder abuse will be highlighted. Interventions should take into account elderly victims' vulnerability, which may be due to physical and/or psychological dependence. The effectiveness of current interventions is limited due to a lack of cohesion and the absence of a specific legal framework. The term 'vulnerable adults' is broader than 'elder abuse'. It is difficult to estimate the extent of the problem of elder abuse because of the usually intimate settings within which it takes place - this may affect the strength of the evidence. The purpose of this paper is to encourage a debate on the effectiveness of current legal responses to elder abuse. The true figures of the incidence of abuse may never be known. Consequently, in criminal law trials, providing evidence on abuse can be very traumatic and intimidating for a victim because of the circumstances in which it generally takes place. The irony is that the likelihood of a criminal prosecution may be higher in more serious types of abuse where the evidence is overwhelming, on the basis of the res ipsa loquitor (facts speak for themselves) principle. The abuse in question may be domestic or institutional; the effects nevertheless are the same.


Identification of Monilinia fructigena, M. fructicola, M. laxa, and Monilia polystroma on Inoculated and Naturally Infected Fruit Using Multiplex PCR.
Monilinia fructigena, M. fructicola, M. laxa, and Monilia polystroma each have a different regulatory status. To monitor imported and exported fruit for the presence of quarantined Monilinia or Monilia species, a timely identification method is required. Random amplified polymorphic DNA analysis was used to generate an M. fructigena-specific band that was characterized by sequencing. Using the sequence obtained, primers were designed to amplify bands in the same genomic region of M. fructicola and M. laxa. These bands were also characterized by sequencing. From all three sequences, a multiplex polymerase chain reaction (PCR) method based on a common reverse primer (MO368-5) and three species-specific forward primers (MO368-8R, MO368-10R, and Laxa-R2) was established for the differentiation of the three Monilinia species. The multiplex PCR was tested with additional isolates and consistently produced a 402-bp PCR product for M. fructigena, a 535-bp product for M. fructicola, and a 351-bp product for M. laxa. The method was also used with isolates of the recently characterized Monilia polystroma, and all isolates amplified a 425-bp PCR product. The identification method was shown to amplify a PCR product directly from inoculated apples, and the PCR band produced was specific to the inoculated Monilinia or Monilia species. Furthermore, the multiplex PCR was used to identify Monilinia species on naturally infected stone fruits. The method correctly identified infections by both M. laxa and M. fructicola by successful amplification of corresponding PCR products for each species.


Objective Perfusion Assessment in Gracilis Muscle Interposition-A Novel Software-Based Approach to Indocyanine Green Derived Near-Infrared Fluorescence in Reconstructive Surgery.
Gracilis muscle interposition (GMI) is an established treatment option for complex perineal fistulas and reconstruction. The outcome is limited by complications such as necrosis, impaired wound healing and fistula persistence or recurrence. Quantifiable methods of assessing muscle flap perfusion intraoperatively are lacking. This study evaluates a novel and objective software-based assessment of indocyanine green near-infrared fluorescence (ICG-NIRF) in GMI. Intraoperative ICG-NIRF visualization data of five patients with inflammatory bowel disease (IBD) undergoing GMI for perineal fistula and reconstruction were analyzed retrospectively. A new software was utilized to generate perfusion curves for the specific regions of interest (ROIs) of each GMI by depicting the fluorescence intensity over time. Additionally, a pixel-to-pixel and perfusion zone analysis were performed. The findings were correlated with the clinical outcome. Four patients underwent GMI without postoperative complications within 3 months. The novel perfusion indicators identified here (shape of the perfusion curve, maximum slope value, distribution and range) indicated adequate perfusion. In one patient, GMI failed. In this case, the perfusion indicators suggested impaired perfusion. We present a novel, software-based approach for ICG-NIRF perfusion assessment, identifying previously unknown objective indicators of muscle flap perfusion. Ready for intraoperative real-time use, this method has considerable potential to optimize GMI surgery in the future.


Diversification versus specialization in complex ecosystems.
By analyzing the distribution of revenues across the production sectors of quoted firms we suggest a novel dimension that drives the firms diversification process at country level. Data show a non trivial macro regional clustering of the diversification process, which underlines the relevance of geopolitical environments in determining the microscopic dynamics of economic entities. These findings demonstrate the possibility of singling out in complex ecosystems those micro-features that emerge at macro-levels, which could be of particular relevance for decision-makers in selecting the appropriate parameters to be acted upon in order to achieve desirable results. The understanding of this micro-macro information exchange is further deepened through the introduction of a simplified dynamic model.


Female monopolization mediates the relationship between pre- and postcopulatory sexual traits.
Theory predicts a trade-off between investments in precopulatory (ornaments and armaments) and postcopulatory (testes and ejaculates) sexual traits due to the costs associated with their growth and maintenance within the finite energy resources available. Empirical studies, however, have revealed considerable inconsistency in the strength and direction of relationships among these sexual traits. Ambiguity may result from variance in the marginal benefits gained by increasing investments in either pre- or postcopulatory sexual traits. Here, in a broad comparative study, we test the prediction that the relationship between pre- and postcopulatory sexual traits differs among taxa relative to the importance of male-male contest competition within them. We find that covariance between pre- and postcopulatory sexual traits gradually shifts from strongly positive to strongly negative with increasing male-male contest competition. Thus, our findings reveal a potentially unifying explanation for the oftentimes inconsistent relationships in the strength and direction of covariance among sexual traits.


Evaluation of a Diabetes Education Call Center intervention.
Patients require education and information as they engage in self-help, self-care, and disease management activities. The purpose of this study was to determine how effective voice technologies are in diabetes patient education. A pretest-posttest study was conducted to evaluate the effectiveness of prerecorded educational messages delivered via the telephone to participants with diabetes. The intervention consisted of 24 four-minute messages on the topics of knowledge and prevention, glucose level, diet and activity, and management and coping. Eighteen persons with diabetes participated in the pretest-posttest trial. A total of 324 educational messages were listened to over a 12-week intervention period. The pretest-posttest trial demonstrated that a brief telephone-based diabetes education intervention can have a significant impact on increasing frequency of checking blood for glucose (p = 0.017), improving general diabetes knowledge (p = 0.048), and improving insulin-specific knowledge (p = 0.020). Automated educational interventions should be based on scientifically sound evidence and can be effectively delivered by telephone. Automated telephone-based diabetes education may be used alone or as a supplement to existing diabetes education. Automated education is a viable solution when healthcare organizations and regions that as a result of a lack of human and financial resources cannot afford a diabetes educator.


[Role of biomarkers of acute kidney damage during lithotripsy of high-density stones].
To obtain the information about functional state of kidneys in patients with urolithiasis before and after treatment, as well as to study the damaging effect of different types of energy used for fragmentation of high-density stones. A total of 105 patients aged from 25 to 62 years with high-density stones were undergone to lithotripsy. In Group 1 (n=38), Group 2 (n=32) and Group 3 (n=35) contact laser lithotripsy, contact ultrasound lithotripsy and extracorporeal shock-wave lithotripsy was used, respectively. In all cases the clinical and biochemical blood and urine tests were performed as well as leukocyte migration inhibition test, selective proteinuria, a urine level of inteleukin-18 (IL-18) and urine NGAL (lipocalin-2) were assessed. The first examination was done the day before lithotripsy and the next ones were performed after 3 hours, on the 1st and 5th day after the intervention. In all cases dense unilateral kidney stones of size 0.8-2 cm were detected. The stone-free rate after contact lithotripsy was 92.8%. After ESWL, the stone-free rate after two weeks was 94.9%. The average duration of lithotripsy in the Group 1, 2 and 3 was 40+/-3.8 min, 35+/-2.3 min and 32+/-3.6 min, respectively. Based on the level of biomarkers of AKI, laser lithotripsy allows to achieve stone fragmentation with the least damage. Our study proves that IL-18, NGAL, leukocyte migration inhibition test and selective proteinuria allows to diagnose AKI at early stages, as well as to objectively assess the functional state of the kidneys after lithotripsy. The obtained data proves that laser lithotripsy is the safest method as assessed by damaging effects on the kidney parenchyma.


Ocular attacks after phacoemulsification and intraocular lens implantation in patients with Behçet disease.
To elucidate factors related to ocular inflammatory attacks after cataract surgery, limited to a single procedure of phacoemulsification and intraocular lens implantation, in patients with Behçet disease. This retrospective study included 12 consecutive patients (16 eyes) with Behçet disease, who underwent phacoemulsification and intraocular lens implantation during 4 years from January 1995 to December 1998 at three institutions. Their medical records were reviewed, and factors related to the ocular attacks were analyzed. Four eyes of 3 patients experienced ocular attacks during 1 year before cataract surgery, while 4 eyes of 4 patients developed ocular attacks during 1 year after the surgery. The development of ocular attacks after cataract surgery was significantly related with the presence of ocular attacks during 1 year before the surgery (p = 0.0286, chi(2) test). The patients' age or gender, the duration of Behçet disease or oral medications for Behçet disease did not show any relationship with the presence or absence of ocular attacks after cataract surgery. The visual acuity improved in all patients after the surgery, including those who developed ocular attacks. The experience of ocular attacks during 1 year before cataract surgery is related to postoperative ocular attacks. Despite postoperative ocular attacks, phacoemulsification with intraocular lens implantation is a safe procedure to expect a good visual outcome in patients with Behçet disease.


Public long-term care insurance: a way to ensure sustainable continuity of care for frail older people.
A necessary component of the prescribed continuing care act should be the creation of a long-term care insurance program in Canada. This social innovation has been implemented in many countries in recent decades to address the aging of the population and associated pandemic of chronic diseases. A specific autonomy insurance fund would have to be created and funded through transfers of current funding for long-term care, tax credits for disabled individuals and substantial new investment to make the shift from institutional to home care. Following a standardized assessment by case managers, an autonomy support benefit would be determined to fund "in kind" public or private services or a "cash-for-care" allowance for older people to purchase services. Case managers would be responsible for coordinating the care, controlling the quality of services and reassessing needs according to changes in individuals' conditions. Funding reform of long-term care would prioritize home care and optimize the functioning of integrated care to improve support for frail older people.


Postnatal expression in hyaline cartilage of constitutively active human collagenase-3 (MMP-13) induces osteoarthritis in mice.
It has been suggested that increased collagenase-3 (MMP-13) activity plays a pivotal role in the pathogenesis of osteoarthritis (OA). We have used tetracycline-regulated transcription in conjunction with a cartilage-specific promoter to target a constitutively active human MMP-13 to the hyaline cartilages and joints of transgenic mice. Postnatal expression of this transgene resulted in pathological changes in articular cartilage of the mouse joints similar to those observed in human OA. These included characteristic erosion of the articular cartilage associated with loss of proteoglycan and excessive cleavage of type II collagen by collagenase, as well as synovial hyperplasia. These results demonstrate that excessive MMP-13 activity can result in articular cartilage degradation and joint pathology of the kind observed in OA, suggesting that excessive activity of this proteinase can lead to this disease.


Chemoprevention of 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster cheek pouch carcinogenesis by a 5-lipoxygenase inhibitor, garcinol.
Our previous studies have shown that aberrant arachidonic acid metabolism, especially the 5-lipoxygenase (5-Lox) pathway, is involved in oral carcinogenesis and can be targeted for cancer prevention. To develop potent topical agents for oral cancer chemoprevention, 5 known 5-Lox inhibitors from dietary and synthetic sources (Zileuton, ABT-761, licofelone, curcumin, and garcinol) were evaluated in silico for their potential efficacy. Garcinol, a polyisoprenylated benzophenone from the fruit rind of Garcinia spp., was found to be a promising agent based on the calculation of a theoretical activity index. Computer modeling showed that garcinol well fit the active site of 5-Lox, and potentially inhibited enzyme activity through interactions between the phenolic hydroxyl groups and the non-heme catalytic iron. In a short-term study on 7,12-dimethylbenz[a]anthracene (DMBA)-treated hamster cheek pouch, topical garcinol suppressed leukotriene B4 (LTB4) biosynthesis and inhibited inflammation and cell proliferation in the oral epithelium. In a long-term carcinogenesis study, topical garcinol significantly reduced the size of visible tumors, the number of cancer lesions, cell proliferation, and LTB4 biosynthesis. These results demonstrated that topical application of a 5-Lox inhibitor, garcinol, had chemopreventive effect on DMBA-induced hamster cheek pouch carcinogenesis.


Impact of Ebola experiences and risk perceptions on mental health in Sierra Leone, July 2015.
The mental health impact of the 2014-2016 Ebola epidemic has been described among survivors, family members and healthcare workers, but little is known about its impact on the general population of affected countries. We assessed symptoms of anxiety, depression and post-traumatic stress disorder (PTSD) in the general population in Sierra Leone after over a year of outbreak response. We administered a cross-sectional survey in July 2015 to a national sample of 3564 consenting participants selected through multistaged cluster sampling. Symptoms of anxiety and depression were measured by Patient Health Questionnaire-4. PTSD symptoms were measured by six items from the Impact of Events Scale-revised. Relationships among Ebola experience, perceived Ebola threat and mental health symptoms were examined through binary logistic regression. Prevalence of any anxiety-depression symptom was 48% (95% CI 46.8% to 50.0%), and of any PTSD symptom 76% (95% CI 75.0% to 77.8%). In addition, 6% (95% CI 5.4% to 7.0%) met the clinical cut-off for anxiety-depression, 27% (95% CI 25.8% to 28.8%) met levels of clinical concern for PTSD and 16% (95% CI 14.7% to 17.1%) met levels of probable PTSD diagnosis. Factors associated with higher reporting of any symptoms in bivariate analysis included region of residence, experiences with Ebola and perceived Ebola threat. Knowing someone quarantined for Ebola was independently associated with anxiety-depression (adjusted OR (AOR) 2.3, 95% CI 1.7 to 2.9) and PTSD (AOR 2.095% CI 1.5 to 2.8) symptoms. Perceiving Ebola as a threat was independently associated with anxiety-depression (AOR 1.69 95% CI 1.44 to 1.98) and PTSD (AOR 1.86 95% CI 1.56 to 2.21) symptoms. Symptoms of PTSD and anxiety-depression were common after one year of Ebola response; psychosocial support may be needed for people with Ebola-related experiences. Preventing, detecting, and responding to mental health conditions should be an important component of global health security efforts.


Myocardial Hypertrophic Preconditioning Attenuates Cardiomyocyte Hypertrophy and Slows Progression to Heart Failure Through Upregulation of S100A8/A9.
Transient preceding brief ischemia provides potent cardioprotection against subsequent long ischemia, termed ischemic preconditioning. Here, we hypothesized that transient short-term hypertrophic stimulation would induce the expression of hypertrophy regression genes and render the heart resistant to subsequent hypertrophic stress, and slow the progression to heart failure, as well. Cardiomyocyte hypertrophy was induced in mice by either transverse aortic constriction or an infusion of phenylephrine, and in neonatal rat ventricular cardiomyocytes by norepinephrine exposures. In the preconditioning groups, hypertrophic stimulation was provided for 1 to 7 days and then withdrawn for several days by either aortic debanding or discontinuing phenylephrine or norepinephrine treatment, followed by subsequent reexposure to the hypertrophic stimulus for the same period as in the control group. One or 6 weeks after transverse aortic constriction, the heart weight/body weight ratio was lower in the preconditioning group than in the control group, whereas the lung weight/body weight ratio was significantly decreased 6 weeks after transverse aortic constriction. Similar results were obtained in mice receiving phenylephrine infusion and neonatal rat ventricular cardiomyocytes stimulated with norepinephrine. Both mRNA and protein expression of S100A8 and S100A9 showed significant upregulation after the removal of hypertrophic stimulation and persisted for 6 weeks in response to reimposition of transverse aortic constriction. The treatment with recombinant S100A8/A9 inhibited norepinephrine-induced myocyte hypertrophy and reduced the expression of calcineurin and NFATc3, but the silencing of S100A8/A9 prevented such changes. Preconditioning with prohypertrophic factors exerts an antihypertrophic effect and slows the progression of heart failure, indicating the existence of the phenomenon for hypertrophic preconditioning.


An approach to supervision of students on fieldwork.
This paper describes a method for supervision of occupational therapy students on clinical affiliation. The approach is based on the assumption that students behave in ways that are consistent with Levels 3 through 4 of Jane Loevinger's stages of personality development. The most effective supervisory approach for students at each level of behavior is described. The model assumes that the level of student can be matched with a supervisory approach to result in a more satisfying experience for both student and supervisor.


Potential for Biocontrol of Hairy Root Disease by a Paenibacillus Clade.
Rhizogenic Agrobacterium biovar 1 is the causative agent of hairy root disease (HRD) in the hydroponic cultivation of tomato and cucumber causing significant losses in marketable yield. In order to prevent and control the disease chemical disinfectants such as hydrogen peroxide or hypochlorite are generally applied to sanitize the hydroponic system and/or hydroponic solution. However, effective control of HRD sometimes requires high disinfectant doses that may have phytotoxic effects. Moreover, several of these chemicals may be converted to unwanted by-products with human health hazards. Here we explored the potential of beneficial bacteria as a sustainable means to control HRD. A large collection of diverse bacterial genera was screened for antagonistic activity against rhizogenic Agrobacterium biovar 1 using the agar overlay assay. Out of more than 150 strains tested, only closely related Paenibacillus strains belonging to a particular clade showed antagonistic activity, representing the species P. illinoisensis, P. pabuli, P. taichungensis, P. tundrae, P. tylopili, P. xylanexedens, and P. xylanilyticus. Assessment of the spectrum of activity revealed that some strains were able to inhibit the growth of all 35 rhizogenic agrobacteria strains tested, while others were only active against part of the collection, suggesting a different mode of action. Preliminary characterization of the compounds involved in the antagonistic activity of two closely related Paenibacillus strains, tentatively identified as P. xylanexedens, revealed that they are water-soluble and have low molecular weight. Application of a combination of these strains in greenhouse conditions resulted in a significant reduction of HRD, indicating the great potential of these strains to control HRD.


Unexpected common mechanistic pathways for embryotoxicity of warfarin and lovastatin.
Novel molecular content for fast in vitro strategies in the context of safety tests concerning developmental toxicity has a potential to substantially reduce animal experiments according to the "3R" concept (Reduce/Refine/Replace). Here we present and discuss data from a differential proteomic profiling of samples generated using embryonic stem cell derived in vitro models treated with a set of model substances. Among substance-dependent proteomic changes, potential surrogate markers were some isoforms of heat shock proteins and a component of the Ras pathway, present in several redundant isoforms due to posttranslational modifications. Both proteins are implicated in cell migration, cell survival, growth and embryonic development. Using the examples of warfarin and lovastatin, two substances with entirely different primary targets, the surrogate marker signature nevertheless indicates a common embryotoxic mode of action. We discuss these findings observed in in vitro toxicity tests, in a context of clinical validation and evidence-based toxicology.


Transmission Dynamics of Hyper-Endemic Multi-Drug Resistant Klebsiella pneumoniae in a Southeast Asian Neonatal Unit: A Longitudinal Study With Whole Genome Sequencing.
Background:Klebsiella pneumoniae is an important and increasing cause of life-threatening disease in hospitalized neonates. Third generation cephalosporin resistance (3GC-R) is frequently a marker of multi-drug resistance, and can complicate management of infections. 3GC-R K. pneumoniae is hyper-endemic in many developing country settings, but its epidemiology is poorly understood and prospective studies of endemic transmission are lacking. We aimed to determine the transmission dynamics of 3GC-R K. pneumoniae in a newly opened neonatal unit (NU) in Cambodia and to address the following questions: what is the diversity of 3GC-R K. pneumoniae both within- and between-host; to what extent is high carriage prevalence driven by ward-based transmission; and to what extent can environmental contamination explain patterns of patient acquisition. Methods: We performed a prospective longitudinal study between September and November 2013. Rectal swabs from consented patients were collected upon NU admission and every 3 days thereafter. Morphologically different colonies from swabs growing cefpodoxime-resistant K. pneumoniae were selected for whole-genome sequencing (WGS). Results: One hundred and fifty-eight samples from 37 patients and 7 environmental sites were collected. 32/37 (86%) patients screened positive for 3GC-R K. pneumoniae and 93 colonies from 119 swabs were successfully sequenced. Isolates were resistant to a median of six (range 3-9) antimicrobials. WGS revealed high diversity; pairwise distances between isolates from the same patient were either 0-1 SNV or >1,000 SNVs; 19/32 colonized patients harbored K. pneumoniae colonies differing by >1000 SNVs. Diverse lineages accounted for 18 probable importations to the NU and nine probable transmission clusters involving 19/37 (51%) of screened patients. Median cluster size was five patients (range 3-9). Seven out of 46 environmental swabs (15%) were positive for 3GC-R K. pneumoniae. Environmental sources were plausible sources for acquisitions in 2/9 transmission clusters, though in both cases other patients were also plausible sources. Conclusion: The epidemiology of 3GC-R K. pneumoniae was characterized by multiple introductions, high within- and between host diversity and a dense network of cross-infection, with half of screened neonates part of a transmission cluster. We found no evidence to suggest that environmental contamination was playing a dominant role in transmission.


Hormonal status and ICU-acquired paresis in critically ill patients.
The pathogenesis of intensive care unit-acquired paresis (ICUAP), a frequent and severe complication of critical illness, is poorly understood. Since ICUAP has been associated with female gender in some studies, we hypothesized that hormonal dysfunction might contribute to ICUAP. To determine the relationship between hormonal status, ICUAP and mortality in patients with protracted critical illness. Prospective observational study. Four medical and surgical ICUs. ICU patients mechanically ventilated for >7 days. None. Plasma levels of insulin growth factor-1 (IgF1), prolactin, thyroid stimulating hormone (TSH), follicular stimulating hormone (FSH), luteinizing hormone (LH), estradiol, progesterone, testosterone, dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulphate (DHEAS) and cortisol were measured on the first day patients were awake (day 1). Mean blood glucose from admission to day 1 was calculated. ICUAP was defined as Medical Research Council sum score <48/60 on day 7. We studied 102 patients (65 men and 37 women, 29 post-menopausal), of whom 24 (24%) died during hospitalization. Among the 86 patients tested at day 7, 39 (49%) had ICUAP, which was more frequent in women (63% versus men 36%, p = 0.02). Mean blood glucose was higher in patients with ICUAP. Estradiol/testosterone ratio was greater in men with ICUAP. ICUAP 7 days after awakening was associated with increased blood glucose and with biological evidence of hypogonadism in men, while an association with hormonal dysfunction was not detected in women.


Delineating the psychiatric and behavioral phenotype of recurrent 2q13 deletions and duplications.
Recurrent deletions and duplications at the 2q13 locus have been associated with developmental delay (DD) and dysmorphisms. We aimed to undertake detailed clinical characterization of individuals with 2q13 copy number variations (CNVs), with a focus on behavioral and psychiatric phenotypes. Participants were recruited via the Unique chromosomal disorder support group, U.K. National Health Service Regional Genetics Centres, and the DatabasE of genomiC varIation and Phenotype in Humans using Ensembl Resources (DECIPHER) database. A review of published 2q13 patient case reports was undertaken to enable combined phenotypic analysis. We present a new case series of 2q13 CNV carriers (21 deletion, 4 duplication) and the largest ever combined analysis with data from published studies, making a total of 54 deletion and 23 duplication carriers. DD/intellectual disabilities was identified in the majority of carriers (79% deletion, 70% duplication), although in the new cases 52% had an IQ in the borderline or normal range. Despite the median age of the new cases being only 9 years, 64% had a clinical psychiatric diagnosis. Combined analysis found attention deficit hyperactivity disorder (ADHD) to be the most frequent diagnosis (48% deletion, 60% duplication), followed by autism spectrum disorders (33% deletion, 17% duplication). Aggressive (33%) and self-injurious behaviors (33%) were also identified in the new cases. CNVs at 2q13 are typically associated with DD with mildly impaired intelligence, and a high rate of childhood psychiatric diagnoses-particularly ADHD. We have further characterized the clinical phenotype related to imbalances of the 2q13 region and identified it as a region of interest for the neurobiological investigation of ADHD.


Single-cell Profiles and Prognostic Impact of Tumor-Infiltrating Lymphocytes Coexpressing CD39, CD103, and PD-1 in Ovarian Cancer.
Tumor-infiltrating lymphocytes (TIL) are strongly associated with survival in most cancers; however, the tumor-reactive subset that drives this prognostic effect remains poorly defined. CD39, CD103, and PD-1 have been independently proposed as markers of tumor-reactive CD8+ TIL in various cancers. We evaluated the phenotype, clonality, and prognostic significance of TIL expressing various combinations of these markers in high-grade serous ovarian cancer (HGSC), a malignancy in need of more effective immunotherapeutic approaches. Expression of CD39, CD103, PD-1, and other immune markers was assessed by high-dimensional flow cytometry, single-cell sequencing, and multiplex immunofluorescence of primary and matched pre/post-chemotherapy HGSC specimens. Coexpression of CD39, CD103, and PD-1 ("triple-positive" phenotype) demarcated subsets of CD8+ TIL and CD4+ regulatory T cells (Treg) with a highly activated/exhausted phenotype. Triple-positive CD8+ TIL exhibited reduced T-cell receptor (TCR) diversity and expressed genes involved in both cytolytic and humoral immunity. Triple-positive Tregs exhibited higher TCR diversity and a tumor-resident phenotype. Triple-positive TIL showed superior prognostic impact relative to TIL expressing other combinations of these markers. TIGIT was uniquely upregulated on triple-positive CD8+ effector cells relative to their CD4+ Treg counterparts. Coexpression of CD39, CD103, and PD-1 demarcates highly activated CD8+ and CD4+ TIL with inferred roles in cytolytic, humoral, and regulatory immune functions. Triple-positive TIL demonstrate exceptional prognostic significance and express compelling targets for combination immunotherapy, including PD-1, CD39, and TIGIT.


Edge detecting for range data using Laplacian operators.
Feature extraction in image data has been investigated for many years, and more recently the problem of processing images containing irregularly distributed data has become prominent. Range data are now commonly used in the areas of image processing and computer vision. However, due to the data irregularity found in range images that occurs with a variety of image sensors, direct image processing, in particular edge detection, is a non-trivial problem. Typically, irregular range data would require to be interpolated to a regular grid prior to processing. One example of an edge detection technique than can be directly applied to range images is the scan-line approximation, but this does not employ exact data locations. Therefore, we present novel Laplacian operators that can be applied directly to irregularly distributed data, and in particular we focus on application to irregularly distributed 3D range data for the purpose of edge detection. Within the data distribution framework commonly occurring in range data acquisition devices, our results illustrate that the approach works well over a range of levels of irregularity of data distribution. The use of Laplacian operators on range data is also found to be much less susceptible to noise than the traditional use of Laplacian operators on intensity images.


The LIM-only protein Lmo2 is a bridging molecule assembling an erythroid, DNA-binding complex which includes the TAL1, E47, GATA-1 and Ldb1/NLI proteins.
The LIM-only protein Lmo2, activated by chromosomal translocations in T-cell leukaemias, is normally expressed in haematopoiesis. It interacts with TAL1 and GATA-1 proteins, but the function of the interaction is unexplained. We now show that in erythroid cells Lmo2 forms a novel DNA-binding complex, with GATA-1, TAL1 and E2A, and the recently identified LIM-binding protein Ldb1/NLI. This oligomeric complex binds to a unique, bipartite DNA motif comprising an E-box, CAGGTG, followed approximately 9 bp downstream by a GATA site. In vivo assembly of the DNA-binding complex requires interaction of all five proteins and establishes a transcriptional transactivating complex. These data demonstrate one function for the LIM-binding protein Ldb1 and establish a function for the LIM-only protein Lmo2 as an obligatory component of an oligomeric, DNA-binding complex which may play a role in haematopoiesis.


Intravitreal Bevacizumab for Traumatic Choroidal Rupture.
To report a case of visual loss associated with traumatic choroidal rupture after blunt ocular trauma that was successfully treated with an early intravitreal bevacizumab injection despite the absence of choroidal neovascularization (CNV). A 14-year-old boy presented with visual disturbance in his left eye after sustaining an ocular contusion 4 weeks earlier. The best-corrected visual acuity (BCVA) in the left eye was 20/50. Funduscopic examination revealed macular choroidal rupture accompanied by subretinal hemorrhage. Optical coherence tomography (OCT) showed accumulation of subretinal fluid around a disrupted retinal pigment epithelium/Bruch membrane complex extending into the juxtafoveolar area, but there was no active leakage suggestive of CNV on fluorescein angiography. Intravitreal bevacizumab (1.25 mg) injection was performed to treat persistent serous retinal detachment at macula causing visual loss. There was a reduction of subretinal fluid and concomitant improvement of BCVA to 20/30 within 1 week after intravitreal bevacizumab injection. The BCVA recovered to 20/25 in the left eye after 4 weeks, and only a minimal amount of residual fluid remained according to OCT. Complete resolution of subretinal fluid was observed by OCT at the 6-week follow-up examination, and BCVA improved to 20/20. Good visual acuity (20/20) and stable macula were maintained in the left eye at 1 year of follow-up without recurrence of subretinal fluid accumulation or hemorrhage and CNV. There were no ocular or systemic complications associated with intravitreal bevacizumab injection. Early intravitreal bevacizumab injection could be an effective treatment option for patients with vision loss associated with traumatic choroidal rupture and subretinal fluid within the posterior pole before development of CNV.


Whose turn is it anyway? Latency and the organization of turn-taking in video-mediated interaction.
Latency in video-mediated interaction can frustrate smooth turn-taking: it may cause participants to perceive silence at points where talk should occur, it may cause them to talk in overlap, and it impedes their ability to return to one-speaker-at-a-time. Whilst potentially frustrating for participants, this makes video-mediated interaction a perspicuous setting for the study of social interaction: it is an environment that nurtures the occurrence of turn-taking problems. For this paper, we conducted secondary analysis of 25 video consultations recorded for heart failure, (antenatal) diabetes, and cancer services in the UK. By comparing video recordings of the patient's and clinician's side of the call, we provide a detailed analysis of how latency interferes with the turn-taking system, how participants understand problems, and how they address them. We conclude that in our data latency unnoticed until it becomes problematic: participants act as if they share the same reality.


An emotional processing writing intervention and heart rate variability: the role of emotional approach.
Expressing and understanding one's own emotional responses to negative events, particularly those that challenge the attainment of important life goals, is thought to confer physiological benefit. Individual preferences and/or abilities in approaching emotions might condition the efficacy of interventions designed to encourage written emotional processing (EP). This study examines the physiological impact (as indexed by heart rate variability (HRV)) of an emotional processing writing (EPW) task as well as the moderating influence of a dispositional preference for coping through emotional approach (EP and emotional expression (EE)), in response to a laboratory stress task designed to challenge an important life goal. Participants (n = 98) were randomly assigned to either EPW or fact control writing (FCW) following the stress task. Regression analyses revealed a significant dispositional EP by condition interaction, such that high EP participants in the EPW condition demonstrated higher HRV after writing compared to low EP participants. No significant main effects of condition or EE coping were observed. These findings suggest that EPW interventions may be best suited for those with preference or ability to process emotions related to a stressor or might require adaptation for those who less often cope through emotional approach.


Origin of human T-lymphotrophic virus I-positive T cell lines in adult T cell leukemia. Analysis of T cell receptor gene rearrangement.
Using the clone-specific rearrangement of the T cell receptor gene as the genetic marker of the clonotype, we analyzed the clonal origin of the interleukin 2 (IL-2)-dependent human T-lymphotrophic virus I (HTLV-I)-positive T cell lines established from various adult T cell leukemia (ATL) patients. From a patient with chronic ATL, whose leukemic cells proliferated in vitro in response to IL-2, we repeatedly established leukemic T cell clones having the same rearrangement profile of the T beta chain gene as the leukemic cells. By contrast, established cell lines from acute ATL patients had different beta chain gene rearrangements from those of the leukemic cells. These HTLV-I+ T cell lines might not be the direct progeny of the leukemic cells, but that of T cells infected either in vivo or in vitro. These IL-2-reactive nonleukemic T cells might have been selected in vitro, because their leukemic cells failed to respond to IL-2, despite the expression of IL-2 receptor. The analysis of the T cell receptor gene rearrangement may give a new approach for the elucidation of the mechanism of leukemogenesis and the origin of the HTLV-I+ T cell lines in ATL.


SKH-1 mouse skin painting: a short-term assay to evaluate the tumorigenic activity of cigarette smoke condensate.
The hairless mouse strain SKH-1 was investigated in a short-term assay to assess the tumorigenic activity of mainstream cigarette smoke condensate (CSC). The design chosen was the two-stage dermal tumorigenicity assay (skin painting), with tumor initiation using a single dermal application of the carcinogen/mutagen dimethylbenz(a)anthracene applied to the back at a non-tumorigenic dose and tumor promotion by repeated dermal applications of CSC at the same site. The mice reproducibly developed skin tumors at the application site after 15 weeks at a frequency that allowed comparison of the treatment groups. The histopathological examination revealed the benign nature of the tumors (keratotic papillomas). Prolongation of the application period to 25 weeks resulted in the development of malignant tumors (carcinomas), indicating that the benign tumors after 15 weeks can be taken as surrogate endpoints for malignancies progressing after further treatment. After 15 weeks, tumor incidence (the percentage of tumor-bearing mice), tumor multiplicity (the number of tumors per mouse), and tumor-onset (the duration of the application period that resulted in a meaningful tumor incidence) were positively correlated with the initiating and promoting dose. The reproducibility of this assay was at least comparable to that described for other skin painting models in the literature. When the assay system was used to compare CSCs to which different concentrations of benzo(a)pyrene were added, its ability to distinguish these different condensate preparations was comparable to that of other models. The results obtained with the SKH-1 mouse strain suggest its usefulness in the short-term skin painting assay for the comparative investigation of the tumorigenic activity of different CSCs.


Autochthonous Chagas Disease - Missouri, 2018.
On December 13, 2017, the Missouri Department of Health and Senior Services (MDHSS) was notified of a suspected case of Chagas disease in a Missouri woman. The patient had donated blood, and laboratory screening revealed antibodies to Trypanosoma cruzi, the parasite that causes Chagas disease. Evaluation by physicians found no clinical symptoms consistent with Chagas disease. The patient had no travel history that would have suggested a significant risk for Chagas disease risk and had no occupational exposure to the disease agent. She had never received a blood transfusion or organ transplant. Confirmatory testing of the patient's serum at CDC for T. cruzi antibody was consistent with infection. These findings raise the possibility that the exposure to T. cruzi occurred locally (autochthonously) in Missouri. Although the insect vector for the parasite T. cruzi, triatomines (commonly known as "kissing bugs"), has been identified previously in Missouri, no locally acquired human cases of Chagas disease have been identified in the state. Health care providers and public health professionals should be aware of the possibility of locally acquired Chagas disease in the southern United States.


The combination of bendamustine, bortezomib, and rituximab for patients with relapsed/refractory indolent and mantle cell non-Hodgkin lymphoma.
Given the significant activity and tolerability of bendamustine, rituximab, and bortezomib in patients with relapsed indolent and mantle cell non-Hodgkin lymphoma, and laboratory studies suggesting synergistic activity, we conducted a multicenter phase 2 study of the bendamustine/bortezomib/rituximab combination. Patients with relapsed or refractory indolent and mantle cell lymphoma with adequate organ function were treated with bendamustine 90 mg/m² days 1 and 4; rituximab 375 mg/m² day 1, and bortezomib 1.3 mg/m² days 1, 4, 8, 11. Six 28-day cycles were planned. Thirty patients (7 with mantle cell lymphoma) were enrolled and treated. Eight patients experienced serious adverse events, including one event of grade 5 sepsis. Common nonhematologic adverse events were generally grade 1 or grade 2 and included nausea (50%), neuropathy (47%), fatigue (47%), constipation (40%), and fever (40%). Of 29 patients evaluable for efficacy, 24 (83%) achieved an objective response (including 15 with complete response). With median follow-up of 24 months, 2-year progression-free survival is 47% (95% confidence interval, 25%-69%). On the basis of these promising results, the US cooperative groups have initiated randomized trials to evaluate this regimen in follicular and mantle cell lymphoma. This trial was registered at www.clinicaltrials.gov as #NCT00547534.


Assessing Potential Oligomerization Reaction Mechanisms of Isoprene Epoxydiols on Secondary Organic Aerosol.
Extensive studies of secondary organic aerosol (SOA) formation have identified isoprene epoxydiol (IEPOX) intermediates as key species in the formation of isoprene-derived SOA. Recent work has suggested that isoprene-derived dimers and oligomers may constitute a significant fraction of SOA, but a mechanism for the formation of such abundant SOA components has yet to be established. The potential for dimer formation from the nucleophilic addition of 2-methyltetrol to trans-β-IEPOX was assessed through a series of model epoxide-nucleophile experiments using nuclear magnetic resonance (NMR) spectroscopy. These experiments helped establish a rigorous understanding of structural, stereochemical, and NMR chemical shift trends, which were used along with nucleophilic strength calculations to interpret the results of the trans-β-IEPOX + 2-methyltetrol reaction and evaluate its relevance in the atmosphere. A preference for less sterically hindered nucleophiles was observed in all model systems. In all addition products, a significant increase in NMR chemical shift was observed directly adjacent to the epoxide-nucleophile linkage, with smaller decreases in chemical shift at all other sites. A partial NMR assignment of a single trans-β-IEPOX + 2-methyltetrol nucleophilic addition product was obtained, but nucleophilic strength calculations suggest that 2-methyltetrol is a poor nucleophile. Therefore, this reaction is unlikely to significantly contribute to dimer and oligomer formation on SOA. Nevertheless, the structural and stereochemical considerations, NMR assignments, and NMR chemical shift trends reported here will prove useful in future attempts to synthesize dimer and oligomer analytical standards.


Inflammatory response and genotoxicity of seven wood dusts in the human epithelial cell line A549.
Exposure to wood dust is common in many workplaces. Epidemiological studies indicate that occupational exposure to hardwood dusts is more harmful than to softwood dusts. In this study, human epithelial cell line A549 was incubated with well-characterized dusts from six commonly used wood species and from medium density fibreboard (MDF), at concentrations between 10 and 300microg/ml. After 3 and 6h of incubation, genotoxicity was assessed by measurement of DNA damage with the single-cell gel electrophoresis (comet) assay and inflammation was measured by the expression of IL-6 and IL-8 mRNA and by the amount of IL-8 protein. There was a 1.2-1.4-fold increase in DNA strand breaks after incubation with beech, teak, pine and MDF dusts compared with the levels in untreated cells, but after 6h only the increase induced by the MDF dust remained. Increased expression of cellular IL-6 and IL-8 mRNA was induced by all of the wood dusts at both times. Similar to IL-8 mRNA expression, the amounts of secreted IL-8 protein were elevated, except after incubation with oak dust, where a marginal reduction was seen. On the basis of the effects on IL-8 mRNA expression, the wood dusts could be divided into three groups, with teak dust being the most potent, MDF, birch, spruce and pine being intermediate, and beech and oak being the least potent. The induction of DNA strand breaks did not correlate well with the interleukin response. In conclusion, all wood dusts induced cytokine responses, and some dusts induced detectable DNA damage. The inflammatory potency seemed intermediate for dusts from the typical softwoods spruce and pine, whereas the dusts from species linked to cancer, beech and oak, were the least inflammatory. The variation of the effects induced by different wood dusts over time indicates that the DNA damage was not secondary to the cytokine response. Although hardwoods are often considered more harmful than softwoods by regulatory agencies, the current experiments do not provide evidence for a clear-cut distinction between toxicities of hardwood and softwood dust.


The functional interaction between Acyl-CoA synthetase 4, 5-lipooxygenase and cyclooxygenase-2 controls tumor growth: a novel therapeutic target.
The acyl-CoA synthetase 4 (ACSL4), which esterify mainly arachidonic acid (AA) into acyl-CoA, is increased in breast, colon and hepatocellular carcinoma. The transfection of MCF-7 cells with ACSL4 cDNA transforms the cells into a highly aggressive phenotype and controls both lipooxygenase-5 (LOX-5) and cyclooxygenase-2 (COX-2) metabolism of AA, suggesting a causal role of ACSL4 in tumorigenesis. We hypothesized that ACSL4, LOX-5 and COX-2 may constitute potential therapeutic targets for the control of tumor growth. Therefore, the aim of this study was to use a tetracycline Tet-Off system of MCF-7 xenograft model of breast cancer to confirm the effect of ACSL4 overexpression on tumor growth in vivo. We also aim to determine whether a combinatorial inhibition of the ACSL4-LOX-COX-2 pathway affects tumor growth in vivo using a xenograft model based on MDA-MB-231 cells, a highly aggressive breast cancer cell line naturally overexpressing ACSL4. The first novel finding is that stable transfection of MCF-7 cells with ACSL4 using the tetracycline Tet-Off system of MCF-7 cells resulted in development of growing tumors when injected into nude mice. Tumor xenograft development measured in animals that received doxycycline resulted in tumor growth inhibition. The tumors presented marked nuclear polymorphism, high mitotic index and low expression of estrogen and progesterone receptor. These results demonstrate the transformational capacity of ACSL4 overexpression. We examined the effect of a combination of inhibitors of ACSL4, LOX-5 and COX-2 on MDA-MB-231 tumor xenografts. This treatment markedly reduced tumor growth in doses of these inhibitors that were otherwise ineffective when used alone, indicating a synergistic effect of the compounds. Our results suggest that these enzymes interact functionally and form an integrated system that operates in a concerted manner to regulate tumor growth and consequently may be potential therapeutic targets for the control of proliferation as well as metastatic potential of cancer cells.


Smart polymers for controlled delivery of proteins and peptides: a review of patents.
Protein and peptide-based therapeutic agents have unique physiochemical properties such as high molecular weight, short half life, requirement of a sustained plasma level for the desired therapeutic effect, liable to physical and chemical instability by gastric enzymes and harsh acidic environment as well as first pass metabolism, which makes their delivery a challenge. The delivery of proteins and peptides using various routes of administration like oral, nasal, rectal, pulmonary, buccal, vaginal and transdermal route is found to exhibit limitations, poor permeability and degradation being major limitations. Use of parenteral route is found to overcome these problems but patient compliance is poor due to the need for frequent administration. Use of control delivery for these drugs using smart polymers seems promising as they overcome the limitations posed by other routes of delivery. Smart polymers increase patient compliance, maintain stability of the drug, and maintain drug level in therapeutic window and are easy to manufacture. Different types of smart polymer-based delivery systems, such as sensitive to temperature, phase, pH, electric charge, light, and biochemicals, and their application in controlling the release of the incorporated drug to obtain a sustained plasma level has been discussed. Smart polymers, however, face challenges with regard to high burst release, unpredictable behavior in later part of biphasic release profile, overall drug release kinetics, conformational stability during processing, and preserving biological activity after getting released. Several patents overcoming these inherent problems associated with smart polymers have been reviewed. At the end, the future direction and potential of smart polymer-based delivery system for drug delivery has been presented in brief.


CXCL12 expression within the CNS contributes to the resistance against experimental autoimmune encephalomyelitis in Albino Oxford rats.
Experimental autoimmune encephalomyelitis (EAE) is an animal model of multiple sclerosis, a chronic inflammatory and demyelinating disease of the CNS. Albino Oxford (AO) rats are resistant to the induction of EAE, while the disease can be readily induced in Dark Agouti (DA) rats. Here we investigated a potential contribution of the CNS milieu in the limitation of the encephalitogenic autoimmune response. EAE was induced by immunization of the respective rat strains with spinal cord homogenate emulsified in complete Freund's adjuvant. AO rats did not exhibit clinical signs after immunization while DA rats developed severe neurologic deficits. Infiltration of immune cells into spinal cords (SC) was evident in both strains 12-14 days after the immunization. EAE lesions of AO rats contained substantially lower numbers of CD4+ T cells and CD11b+ cells compared to those in DA rats. This went together with lower levels of interferon (IFN)-γ and interleukin (IL)-17 in the cells isolated from SC. We found a dramatic increase of CXCL12 expression in SC tissue and microvessels of AO rats, whereas DA rats markedly decreased the expression of this chemokine within their CNS. Administration of the CXCL12 antagonist AMD3100 to a substrain of AO rats that developed a weak EAE led to earlier onset and exacerbation of the disease. These results suggest a role of CXCL12 in down-regulating autoimmune processes in AO rats during EAE. Therapeutic modulation of CXCL12 could be a promising strategy for the treatment of CNS autoimmunity.


Ultrasound for the evaluation of femoroacetabular impingement of the cam type. Diagnostic performance of qualitative criteria and alpha angle measurements.
To develop and assess a technique to evaluate cam type femoroacetabular impingement (FAI) using ultrasound (US). Fifty patients (24 women, 26 men) were included (mean age: 39.1 years; age range: 16-59). US images of the anterior and anterosuperior contour of the femoral neck were obtained and analysed in 50 patients. Non-spherical shape of the head-neck junction (cam deformity), bony protuberances at the femoral neck, shape of the femoral neck (waist deficiency) and alpha angle were assessed. Magnetic resonance (MR) arthrography served as the standard of reference. Diagnostic performance and receiver operating characteristics (ROC) curves were calculated. Based on MR arthrography 28 patients had cam-type FAI. On US, an anterosuperior cam deformity was seen in 40/44 patients (Reader 1/Reader 2; sensitivity 93%/89%, specificity 36%/14%). A bony protuberance anterosuperiorly in 23/13 patients (sensitivity 71%/32%, specificity 86%/82%) and an anterosuperior waist deficiency in 19/35 patients (sensitivity 25%/54%, specificity 100%/54%). Sensitivity and specificity of the other criteria were lower than 70% (average of Reader 1 & 2). A technique to evaluate cam type FAI using US is presented. The detection of an anterosuperior cam deformity is sensitive, and presence of an anterosuperior bony protuberance is specific for cam FAI. Alpha angle measurements are not helpful in establishing the diagnosis.


Diversity of Cryptosporidium in brush-tailed rock-wallabies (Petrogale penicillata) managed within a species recovery programme.
Host-parasite relationships are likely to be impacted by conservation management practices, potentially increasing the susceptibility of wildlife to emerging disease. Cryptosporidium, a parasitic protozoan genus comprising host-adapted and host-specific species, was used as an indicator of parasite movement between populations of a threatened marsupial, the brush-tailed rock-wallaby (Petrogale penicillata). PCR screening of faecal samples (n = 324) from seven wallaby populations across New South Wales, identified Cryptosporidium in 7.1% of samples. The sampled populations were characterised as captive, supplemented and wild populations. No significant difference was found in Cryptosporidium detection between each of the three population categories. The positive samples, detected using 18S rRNA screening, were amplified using the actin and gp60 loci. Multi-locus sequence analysis revealed the presence of Cryptosporidium fayeri, a marsupial-specific species, and C. meleagridis, which has a broad host range, in samples from the three population categories. Cryptosporidium meleagridis has not been previously reported in marsupials and hence the pathogenicity of this species to brush-tailed rock-wallabies is unknown. Based on these findings, we recommend further study into Cryptosporidium in animals undergoing conservation management, as well as surveying wild animals in release areas, to further understand the diversity and epidemiology of this parasite in threatened wildlife.


Postoperative patient education: a systematic review.
Knowledge of the effects of the specific approach, mode of delivery, and dose of educational interventions is essential to develop and implement effective postoperative educational interventions. Understanding the relationships of patient characteristics to outcomes is important for educational interventions. The purpose of this systematic review was to examine who would most benefit from postoperative education, given in what type of approach and mode, and at what dose? The sample included 58 studies involving 5,271 participants. Findings indicate that delivery of postoperative patient education through the individualization of content, use of combined media for delivery, provision of education on a one-on-one basis, and in multiple sessions is associated with improvement in educational/health outcomes. Samples that contained individuals younger than 50 years and higher percentages of males showed benefits in outcomes of moderate magnitude. The results highlight the importance of attending to the characteristics of both the elements of postoperative educational interventions and the individual patients in the design and delivery of patient education.


The Contribution of Chemoattractant GPCRs, Formylpeptide Receptors, to Inflammation and Cancer.
A hallmark of inflammatory responses is leukocyte mobilization, which is mediated by pathogen and host released chemotactic factors that activate Gi-protein-coupled seven-transmembrane receptors (GPCRs) on host cell surface. Formylpeptide receptors (FPRs, Fprs in mice) are members of the chemoattractant GPCR family, shown to be critical in myeloid cell trafficking during infection, inflammation, immune responses, and cancer progression. Accumulating evidence demonstrates that both human FPRs and murine Fprs are involved in a number of patho-physiological processes because of their expression on a wide variety of cell types in addition to myeloid cells. The unique capacity of FPRs (Fprs) to interact with numerous structurally unrelated chemotactic ligands enables these receptors to participate in orchestrated disease initiation, progression, and resolution. One murine Fpr member, Fpr2, and its endogenous agonist peptide, Cathelicidin-related antimicrobial peptide (CRAMP), have been demonstrated as key mediators of colon mucosal homeostasis and protection from inflammation and associated tumorigenesis. Recent availability of genetically engineered mouse models greatly expanded the understanding of the role of FPRs (Fprs) in pathophysiology that places these molecules in the list of potential targets for therapeutic intervention of diseases.


Relationship between prion propensity and the rates of individual molecular steps of fibril assembly.
Peptides and proteins possess an inherent propensity to self-assemble into generic fibrillar nanostructures known as amyloid fibrils, some of which are involved in medical conditions such as Alzheimer disease. In certain cases, such structures can self-propagate in living systems as prions and transmit characteristic traits to the host organism. The mechanisms that allow certain amyloid species but not others to function as prions are not fully understood. Much progress in understanding the prion phenomenon has been achieved through the study of prions in yeast as this system has proved to be experimentally highly tractable; but quantitative understanding of the biophysics and kinetics of the assembly process has remained challenging. Here, we explore the assembly of two closely related homologues of the Ure2p protein from Saccharomyces cerevisiae and Saccharomyces paradoxus, and by using a combination of kinetic theory with solution and biosensor assays, we are able to compare the rates of the individual microscopic steps of prion fibril assembly. We find that for these proteins the fragmentation rate is encoded in the structure of the seed fibrils, whereas the elongation rate is principally determined by the nature of the soluble precursor protein. Our results further reveal that fibrils that elongate faster but fracture less frequently can lose their ability to propagate as prions. These findings illuminate the connections between the in vitro aggregation of proteins and the in vivo proliferation of prions, and provide a framework for the quantitative understanding of the parameters governing the behavior of amyloid fibrils in normal and aberrant biological pathways.


How to conduct External Quality Assessment Schemes for the pre-analytical phase?
In laboratory medicine, several studies have described the most frequent errors in the different phases of the total testing process, and a large proportion of these errors occur in the pre-analytical phase. Schemes for registration of errors and subsequent feedback to the participants have been conducted for decades concerning the analytical phase by External Quality Assessment (EQA) organizations operating in most countries. The aim of the paper is to present an overview of different types of EQA schemes for the pre-analytical phase, and give examples of some existing schemes. So far, very few EQA organizations have focused on the pre-analytical phase, and most EQA organizations do not offer pre-analytical EQA schemes (EQAS). It is more difficult to perform and standardize pre-analytical EQAS and also, accreditation bodies do not ask the laboratories for results from such schemes. However, some ongoing EQA programs for the pre-analytical phase do exist, and some examples are given in this paper. The methods used can be divided into three different types; collecting information about pre-analytical laboratory procedures, circulating real samples to collect information about interferences that might affect the measurement procedure, or register actual laboratory errors and relate these to quality indicators. These three types have different focus and different challenges regarding implementation, and a combination of the three is probably necessary to be able to detect and monitor the wide range of errors occurring in the pre-analytical phase.


Identification of drought-responsive microRNAs in Medicago truncatula by genome-wide high-throughput sequencing.
MicroRNAs (miRNAs) are small, endogenous RNAs that play important regulatory roles in development and stress response in plants by negatively affecting gene expression post-transcriptionally. Identification of miRNAs at the global genome-level by high-throughout sequencing is essential to functionally characterize miRNAs in plants. Drought is one of the common environmental stresses limiting plant growth and development. To understand the role of miRNAs in response of plants to drought stress, drought-responsive miRNAs were identified by high-throughput sequencing in a legume model plant, Medicago truncatula. Two hundreds eighty three and 293 known miRNAs were identified from the control and drought stress libraries, respectively. In addition, 238 potential candidate miRNAs were identified, and among them 14 new miRNAs and 15 new members of known miRNA families whose complementary miRNA*s were also detected. Both high-throughput sequencing and RT-qPCR confirmed that 22 members of 4 miRNA families were up-regulated and 10 members of 6 miRNA families were down-regulated in response to drought stress. Among the 29 new miRNAs/new members of known miRNA families, 8 miRNAs were responsive to drought stress with both 4 miRNAs being up- and down-regulated, respectively. The known and predicted targets of the drought-responsive miRNAs were found to be involved in diverse cellular processes in plants, including development, transcription, protein degradation, detoxification, nutrient status and cross adaptation. We identified 32 known members of 10 miRNA families and 8 new miRNAs/new members of known miRNA families that were responsive to drought stress by high-throughput sequencing of small RNAs from M. truncatula. These findings are of importance for our understanding of the roles played by miRNAs in response of plants to abiotic stress in general and drought stress in particular.


Prognostic capacity of automated quantification of suppression time in the EEG of post-asphyctic full-term neonates.
To evaluate the prognostic capacity of a new method for automatic quantification of the length of suppression time in the electroencephalogram (EEG) of a group of asphyxiated newborn infants. Twenty-one full-term newborn infants who had been resuscitated for severe birth asphyxia were studied. Eight channel continuous EEG was recorded for prolonged time periods during the first days of life. Artefact detection or rejection was not applied to the signals. The signals were fed through a pretrained classifier and then segmented into burst and suppression periods. Total suppression length per hour was calculated. All surviving patients were followed with structured neurodevelopmental assessments to at least 18 months of age. The patients who developed neurodevelopmental disability or died had significant suppression periods in their EEG during the first days of life while the patients who had a normal follow-up had no or negligible amount of suppression. This new method for automatic quantification of suppression periods in the raw, neonatal EEG discriminates infants with good from those with poor outcome.


Wavelength reuse in a bidirectional radio-over-fiber link based on cross-gain and cross-polarization modulation in a semiconductor optical amplifier.
Wavelength reuse in a bidirectional radio-over-fiber link is proposed and demonstrated based on the cross-gain modulation (XGM) and the cross-polarization modulation in a semiconductor optical amplifier (SOA). With a polarization beam splitter placed at the remote antenna unit, the polarization-modulated signal generated at the SOA is converted into noninverted and inverted intensity-modulated signals. The noninverted signal is used to cancel the inverted XGM-induced intensity-modulated signal to form a clean optical carrier for wavelength reuse in upstream signal transmission, while the inverted intensity-modulated signal is combined constructively to enhance the XGM-induced signal, providing robust downlink service. A bidirectional transmission of 5 GHz RF signal carrying a 50 MBaud 16 quadrature amplitude modulation baseband signal is experimentally implemented. The error vector magnitude degradation due to the fiber transmission for both the downlink and uplink signal is about 0.2%.


Bullying in schools and exposure to domestic violence.
The study aimed to investigate the relationship between bullying and victimization in school and exposure to interparental violence in a nonclinical sample of Italian youngsters. A cross-sectional study was conducted with a sample of 1059 Italian elementary and middle school students. Participants completed a self-report anonymous questionnaire measuring bullying and victimization and exposure to interparental violence. The questionnaire also included measures on parental child abuse and socio-demographic variables. Almost half of all boys and girls reported different types of bullying and victimization in the previous 3 months, with boys more involved than girls in bullying others. Exposure to interparental physical violence and direct bullying were significantly associated especially for girls: girls exposed to father's violence against the mother and those exposed to mother's violence against the father were among the most likely to bully directly others compared with girls who had not been exposed to any interparental violence. Hierarchical multiple regression analyses revealed that bullying and victimization were predicted by exposure to interparental violence, especially mother-to-father violence, over and above age, gender, and child abuse by the father. Exposure to interparental violence is associated with bullying and victimization in school, even after controlling for direct child abuse. Violence within the family has detrimental effects on the child's behavior; schools, in this regard, can play a fundamental role in early detection of maladjustment.


High density p-type Bi0.5Sb1.5Te3 nanowires by electrochemical templating through ion-track lithography.
High density p-type Bi0.5Sb1.5Te3 nanowire arrays are produced by a combination of electrodeposition and ion-track lithography technology. Initially, the electrodeposition of p-type Bi0.5Sb1.5Te3 films is investigated to find out the optimal conditions for the deposition of nanowires. Polyimide-based Kapton foils are chosen as a polymer for ion track irradiation and nanotemplating Bi0.5Sb1.5Te3 nanowires. The obtained nanowires have average diameters of 80 nm and lengths of 20 microm, which are equivalent to the pore size and thickness of Kapton foils. The nanowires exhibit a preferential orientation along the {110} plane with a composition of 11.26 at.% Bi, 26.23 at.% Sb, and 62.51 at.% Te. Temperature dependence studies of the electrical resistance show the semiconducting nature of the nanowires with a negative temperature coefficient of resistance and band gap energy of 0.089+/-0.006 eV.


Altered balance of proteolytic isoforms of pro-brain-derived neurotrophic factor in autism.
Defects in synaptic development and plasticity may lead to autism. Brain-derived neurotrophic factor (BDNF) plays a critical role in synaptogenesis and synaptic plasticity. BDNF is synthesized as a precursor, pro-BDNF, which can be processed into either a truncated form or into mature BDNF. Previous studies reported increased BDNF-immunoreactive protein in autism, but the mechanism of this increase has not been investigated. We examined BDNF mRNA by real-time reverse transcription-polymerase chain reaction and BDNF protein by Western blotting and enzyme-linked immunosorbent assay in postmortem fusiform gyrus tissue from 11 patients with autism and 14 controls. BDNF mRNA levels were not different in the autism versus control samples, but total BDNF-like immunoreactive protein, measured by enzyme-linked immunosorbent assay, was greater in autism than in controls. Western blotting revealed greater pro-BDNF and less truncated BDNF in autism compared with controls. These data demonstrate that increased levels of BDNF-immunoreactive protein in autism are not transcriptionally driven. Increased pro-BDNF and reduced truncated BDNF are consistent with defective processing of pro-BDNF to its truncated form. Distortion of the balance among the 3 BDNF isoforms, each of which may exhibit different biological activities, could lead to changes in connectivity and synaptic plasticity and, hence, behavior. Thus, imbalance in proteolytic isoforms is a possible new mechanism for altered synaptic plasticity leading to autism.


Elevated expression of caspase-3 inhibitors, survivin and xIAP correlates with low levels of apoptosis in active rheumatoid synovium.
Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) is a tumour necrosis factor (TNF) family member capable of inducing apoptosis in many cell types. Using immunohistochemistry, terminal deoxynucleotidyl transferase biotin-dUTP nick end labelling (TUNEL) and real-time PCR we investigated the expression of TRAIL, TRAIL receptors and several key molecules of the intracellular apoptotic pathway in human synovial tissues from various types of arthritis and normal controls. Synovial tissues from patients with active rheumatoid arthritis (RA), inactive RA, osteoarthritis (OA) or spondyloarthritis (SpA) and normal individuals were studied. Significantly higher levels of TRAIL, TRAIL R1, TRAIL R2 and TRAIL R4 were observed in synovial tissues from patients with active RA compared with normal controls (p < 0.05). TRAIL, TRAIL R1 and TRAIL R4 were expressed by many of the cells expressing CD68 (macrophages). Lower levels of TUNEL but higher levels of cleaved caspase-3 staining were detected in tissue from active RA compared with inactive RA patients (p < 0.05). Higher levels of survivin and x-linked inhibitor of apoptosis protein (xIAP) were expressed in active RA synovial tissues compared with inactive RA observed at both the protein and mRNA levels. This study indicates that the induction of apoptosis in active RA synovial tissues is inhibited despite stimulation of the intracellular pathway(s) that lead to apoptosis. This inhibition of apoptosis was observed downstream of caspase-3 and may involve the caspase-3 inhibitors, survivin and xIAP.


A transient expression assay for the in planta efficacy screening of an antimicrobial peptide against grapevine bacterial pathogens.
Natural and synthetic antimicrobial peptides (AMPs) are of increasing interest as potential resistance conferring elements in plants against pathogen infection. The efficacy of AMPs against pathogens is prescreened by in vitro assays, and promising AMP candidates are introduced as transgenes into plants. As in vitro and in planta environments differ, a prescreening procedure of the AMP efficacy in the plant environment is desired. Here, we report the efficacy of the purified synthetic peptide D4E1 against the grapevine-infecting bacterial pathogens Agrobacterium vitis and Xylophilus ampelinus in vitro and describe for the first time an in planta prescreening procedure based on transiently expressed D4E1. The antimicrobial effect of D4E1 against Ag. vitis and X. ampelinus was shown by a reduction in colony-forming units in vitro in a traditional plate-based assay and by a reduction in bacterial titres in planta as measured by quantitative real-time PCR (qPCR) in grapevine leaves transiently expressing D4E1. A statistically significant reduction in titre was shown for X. ampelinus, but for Ag. vitis, a significant reduction in titre was only observed in a subset of plants. The titres of both grapevine-infecting bacterial pathogens were reduced in an in vitro assay and for X. ampelinus in an in planta assay by D4E1 application. This widens the applicability of D4E1 as a potential resistance-enhancing element to additional pathogens and in a novel plant species. D4E1 is a promising candidate to confer enhanced resistance against the two tested grapevine bacterial pathogens, and the applied transient expression system proved to be a valuable tool for prescreening of D4E1 efficacy in an in planta environment. The described prescreening procedure can be used for other AMPs and might be adapted to other plant species and pathogens before the expensive and tedious development of stably transgenic lines is started.


The influence of smoking on the pattern of disability and relapse risk in AQP4-positive Neuromyelitis Optica Spectrum Disorder, MOG-Ab Disease and Multiple Sclerosis.
the role of smoking on clinical outcomes of central nervous system (CNS) inflammatory disorders is unclear. To assess the effect of smoking on relapses and disability in neuromyelitis optica with aquaporin-4-antibodies (NMOSD-AQP4-Ab), Myelin Oligodendrocyte Glycoprotein-antibodies associated disease (MOGAD) and relapsing remitting Multiple Sclerosis (MS) patients. in a UK cohort of 101 NMOSD-AQP4-Ab, 70 MOGAD and 159 MS, and a Korean cohort of 97 NMOSD-AQ4-Ab, time to first relapse, annualised relapse rate, onset relapse severity and recovery, time to Expanded Disability Status Score(EDSS)/secondary progressive MS (SPMS) were compared between never-smokers and ever-smokers. All clinical data were collected under the local ethics between January 2017 and January 2019. Smoking did not affect the risk of relapse in any of the diseases. The risk of reaching EDSS 6.0 in the UK NMOSD-AQP4-Ab cohort was higher in ever smokers but this did not achieve significance (HR 2.12, p=0.068). When combining the UK and Korea NMOSD-AQP4-Ab cohorts, poorer recovery from the onset attack was significantly more frequent in the ever-smokers versus the never smokers (55% vs 38%, p=0.04). In the MS cohort the risk of reaching EDSS 6 and SPMS was significantly higher in the ever-smokers (HR=2.67, p=0.01 and HR=3.18, p=0.001). In MOGAD similar patterns were seen without reaching significance. In NMOSD-AQP4-Ab smoking associates with worse disability not through an increased risk of relapses but through poor relapse recovery. As in MS, smoking cessation should be encouraged in NMOSD-AQP4-Ab.


Observation of an oxonium ion intermediate in ethanol dehydration to ethene on zeolite.
Zeolite-catalyzed dehydration of ethanol offers promising perspectives for the sustainable production of ethene. Complex parallel-consecutive pathways are proposed to be involved in the reaction network of ethanol dehydration on zeolites, where the initial step of ethanol dehydration is still unclear particularly for the favorable production of ethene at lower temperature. Here we report the observation of a triethyloxonium ion (TEO) in the dehydration of ethanol on zeolite H-ZSM-5 by using ex situ and in situ solid-state NMR spectroscopy. TEO is identified as a stable surface species on the working catalyst, which shows high reactivity during reaction. Ethylation of the zeolite by TEO occurs at lower temperature, leading to the formation of surface ethoxy species and then ethene. The TEO-ethoxide pathway is found to be energetically preferable for the dehydration of ethanol to ethene in the initial stage, which is also supported by theoretical calculations.


Soft occlusal splint therapy in the management of myofascial pain dysfunction syndrome: a follow-up study.
Myofascial Pain Dysfunction Syndrome (MPDS) has been recognized as the most common, nontooth-related chronic orofacial pain condition that confronts dentists. A variety of therapies has been described in literature for its management. The present study is a prospective study carried out to evaluate the efficacy of occlusal splint therapy and compare it with pharmacotherapy (using analgesics and muscle relaxants) in the management of Myofascial Pain Dysfunction Syndrome. Forty patients in the age range of 17-55 years were included in the study and randomly assigned to one of two equally sized groups, A and B. Group A patients received a combination of muscle relaxants and analgesics while Group B patients received soft occlusal splint therapy. All the patients were evaluated for GPI, VAS, maximum comfortable mouth opening, TMJ clicking and tenderness during rest and movement as well as for the number of tender muscles at the time of diagnosis, after the 1 st week of initiation of therapy and every month for three months of follow-up. There was a progressive decrease in GPI scores, number of tender muscles, TMJ clicking and tenderness with various jaw movements and significant improvement in mouth opening in patients on occlusal splint therapy during the follow-up period as compared to the pharmacotherapy group. Occlusal splint therapy has better long-term results in reducing the symptoms of MPDS. It has better patient compliance, fewer side effects, and is more cost-effective than pharmacotherapy; hence, it can be chosen for the treatment of patients with MPDS.


A test of factors mediating the relationship between unwanted sexual activity during childhood and risky sexual practices among women enrolled in the NIMH Multisite HIV Prevention Trial.
This study examined both the direct and indirect associations between unwanted sexual activity during childhood and HIV-related sexual practices of adult women. The sample consisted of 3,346 women recruited from sexually transmitted disease (STD) clinics and health service organizations. The findings demonstrated that participants who reported unwanted sexual activity as a child (USC) were more likely than women who did not report such experience to indicate that they had problems with alcohol, used drugs, received money or drugs in exchange for sex, had unwanted sex, and used mental health services. The women reporting USC also noted a greater number of unprotected sex acts, a greater number of partners, and a greater proportion of sex acts accompanied by drugs or alcohol in the past 90 days. Mediated analyses showed that drug use, exchange of sex for money/drugs, unwanted sex, and to a lesser extent, problems with alcohol mediated the relationship between USC and unprotected sex acts, number of partners, and sex under the influence of drugs and alcohol. These findings suggest that participation in non-sexual risky behaviors among women who report USC may be a bridge to participation in sexual behaviors that increase their risk of HIV infection.


Senescent human periodontal ligament fibroblasts after replicative exhaustion or ionizing radiation have a decreased capacity towards osteoblastic differentiation.
Loss of teeth increases with age or after genotoxic treatments, like head and neck radiotherapy, due to periodontium breakdown. Periodontal ligament fibroblasts represent the main cell type in this tissue and are crucial for the maintenance of homeodynamics and for its regeneration. Here, we have studied the characteristics of human periodontal ligament fibroblasts (hPDLF) that became senescent after replicative exhaustion or after exposure to ionizing radiation, as well as their ability for osteoblastic differentiation. We found that senescent hPDLF express classical markers of senescence, as well as a catabolic phenotype, as shown by the decrease in collagen type I and the increase of MMP-2 expression. In addition, we observed a considerably decreased expression of the major transcription factor for osteoblastic differentiation, i.e. Runx2, a down-regulation which was found to be p53-dependent. In accordance to the above, senescent cells have a significantly decreased alkaline phosphatase gene expression and activity, as well as a reduced ability for osteoblastic differentiation, as found by Alizarin Red staining. Interestingly, cells from both type of senescence express similar characteristics, implying analogous functions in vivo. In conclusion, senescent hPDLF express a catabolic phenotype and express a significantly decreased ability towards an osteoblastic differentiation, thus probably affecting tissue development and integrity.


Mouse embryonic stem cells give rise to gut-like morphogenesis, including intestinal stem cells, in the embryoid body model.
Embryonic stem (ES) cells have been proposed as candidates for cell replacement therapy in patients with intestinal failure because these cells can be expanded indefinitely without losing their pluripotent phenotype. We investigated the differentiation capacity of mouse ES cells into gut-like structures, including intestinal stem cells, and defined culture conditions for efficient induction of formation of these structures. ES cell-derived gut-like structures (ES-guts) were reproducibly induced in developing embryoid bodies (EBs) by day 21 of differentiation culture. ES-guts contained an endodermal epithelium, a smooth muscle layer, interstitial cells of Cajal, and enteric neurons and showed spontaneous contraction. Transplantation of ES-guts under the kidney capsules of immunodeficient mice induced formation of highly differentiated epithelium composed of absorptive cells and goblet cells in the grafts. Immunoreactivity for Musashi-1 (Msi-1), a marker of intestinal stem cells, was detected in 1.9% of the columnar epithelial cells in the graft. Culture with 0.1% dimethyl sulfoxide increased the numbers of ES-guts in EBs, and serum-replacement (SR) culture, in comparison to standard ES culture containing 15% serum, increased the area ratio of ES-guts to EBs. SR culture also promoted maturation of epithelium to form a single layer of columnar epithelial cells, including absorptive cells and goblet cells. Expression of Msi-1 mRNA and protein was significantly enhanced when EBs were cultured under SR conditions. In conclusion, SR conditions efficiently induce formation of ES-guts and promote differentiation of epithelium, including intestinal stem cells. These results suggest the feasibility of cell-based therapy for intestinal failure based on ES cell culture systems.


Economic issues in the development and use of practice guidelines: an application to resource allocation in dentistry.
In this paper, the extent to which practice guidelines using cost-effectiveness data can be used to inform programme decisions is analysed. In particular it is shown that guidelines aimed at informing individual patient-provider decisions are unable to reflect the economic concepts that are required to inform public decisions concerned with making best (i.e., most productive) use of the resources available to serve defined populations. The research on which practice guidelines are based represents an important but incomplete source of information for taking decisions about which clients to serve, with which services, and when in the disease process, in the context of provision of services to groups or populations. Moreover, the inappropriate use of 'individually focused' guidelines to inform 'collectively-focused' decisions can lead to more harm than good. An alternative approach for dealing with the difficult choices faced by decision makers involved in public programmes is identified. An illustration of the proposed approach is presented concerning the provision of pit and fissure sealants to children served by a public health clinic.