Epididymo-testicular anomalies in the undescended testis. How important is their effect on infertility?
Infertility in subjects affected by undesceded testis occurs in from 25 to 100% of the cases according to the various Authors. This depends on whether one or both gonads are concerned and on the age when medical and/or surgical treatment of the condition was begun. Our study was made in two successive periods: first we studied the secretory causes of infertility by examining some histological parameters, (MTD, IFT), and endocrinological ones, (basal testosterone and then after HCG, basal gonadotropin and then after GnRH) in 43 subjects of ages ranging between 2 and 13 years who are affected by uni- or bilateral maldescension. We then analysed the excretory causes of infertility and classified and interpreted 108 epididymo-testicular malformations out of a total of 144 undescended testis observed during our last year of work. We conclude that from a histological point of view, from the third year after birth there is a steady progressive reduction in the tubular fertility index in undescended testis compared to scrotal testis. There is no significant reduction, however, in the mean tubular diameter up to the prepubertal phase. From the endocrinological point of view, we found a normal LH secretion and hypertonia in the pituitary secretion of FSH in bilateral maldescension, not found, however, in unilateral maldescension. Finally, anomalies concerning the epididymo-testicular relationship are found in 75% of undescended testis and of these 36% have a definite effect on infertility of the excretory type.


Bond strength durability of a resin composite on a reinforced ceramic using various repair systems.
This study compared the durability of repair bond strength of a resin composite to a reinforced ceramic after three repair systems. Alumina-reinforced feldspathic ceramic blocks (Vitadur-alpha) (N=30) were randomly divided into three groups according to the repair method: PR-Porcelain Repair Kit (Bisco) [etching with 9.5% hydrofluoric acid+silanization+adhesive]; CJ-CoJet Repair Kit (3M ESPE) [(chairside silica coating with 30microm SiO(2)+silanization (ESPE)-Sil)+adhesive (Visio-Bond)]; CL-Clearfil Repair Kit [diamond surface roughening, etching with 40% H(3)PO(4)+Clearfil Porcelain Bond Activator+Clearfil SE Bond)]. Resin composite was photo-polymerized on each conditioned ceramic block. Non-trimmed beam specimens were produced for the microtensile bond strength (microTBS) tests. In order to study the hydrolytic durability of the repair methods, the beam specimens obtained from each block were randomly assigned to two conditions. Half of the specimens were tested either immediately after beam production (Dry) or after long-term water storage (37 degrees C, 150 days) followed by thermocyling (12,000 cycles, 5-55 degrees C) in a universal testing machine (1mm/min). Failure types were analyzed under an optical microscope and SEM. microTBS results were significantly affected by the repair method (p=0.0001) and the aging conditions (p=0.0001) (two-way ANOVA, Tukey's test). In dry testing conditions, PR method showed significantly higher (p<0.001) repair bond strength (19.8+/-3.8MPa) than those of CJ and CL (12.4+/-4.7 and 9.9+/-2.9, respectively). After long-term water storage and thermocycling, CJ revealed significantly higher results (14.5+/-3.1MPa) than those of PR (12.1+/-2.6MPa) (p<0.01) and CL (4.2+/-2.1MPa) (p<0.001). In all groups when tested in dry conditions, cohesive failure in the composite accompanied with adhesive failure at the interface (mixed failures), was frequently observed (76%, 80%, 65% for PR, CJ and CL, respectively). After aging conditions, while the specimens treated with PR and CJ presented primarily mixed failure types (52% and 87%, respectively), CL group presented mainly complete adhesive failures at the interface (70%). Hydrolytic stability of the repair method based on silica coating and silanization was superior to the other repair strategies for the ceramic tested.


Mechanisms of mitogenic and anti-apoptotic signaling by glucose-dependent insulinotropic polypeptide in beta(INS-1)-cells.
Glucose-dependent insulinotropic polypeptide (GIP) acts as a glucose-dependent growth factor for beta-cells. Here we show that GIP and glucose also act synergistically as anti-apoptotic factors for beta-cells, using the well-differentiated beta-cell line, INS-1. Mitogenic and anti-apoptotic signaling of GIP were dependent upon pleiotropic activation of protein kinase A (PKA)/cAMP regulatory element binder (CREB), mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3-kinase)/PKB signaling modules. The signaling modules activated by GIP were dependent on glucose metabolism and calcium influx and were tightly linked by multiple activating and inhibiting cross-talk. These interactions included: (i) a central role of tyrosine phosphorylation for stimulation of PKA/CREB, MAPK and PI3-kinase/PKB, (ii) inhibition of PKA/CREB by the MAPK pathway at the level of MAPK kinase-1 or downstream, (iii) activation of MAPK signaling by PI3-kinase and PKA at the level of extracellular-signal regulated kinase 1/2 or upstream, and (iv) activation of PKB by MAPK and PKA signaling at the level of PKB or upstream. Furthermore, we demonstrated inhibition of CREB signaling by Ca(2+)/calmodulin kinase I/IV. These results indicated that GIP acts as a mitogenic and anti-apoptotic factor for beta-cells by pleiotropic activation of tightly linked signaling pathways in beta-cells.


Osteomalacia with bone marrow fibrosis due to severe vitamin D deficiency after a gastrointestinal bypass operation for severe obesity.
To present 5 cases of bone biopsy-proven osteomalacia with marrow fibrosis (in 3 cases) after gastric bypass operation, review the relevant literature, and offer preventive strategies. We summarize the clinical presentation, pertinent biochemical and radiologic data, and bone histomorphometric findings in 5 patients, encountered during a period of 17 years, in whom severe vitamin D deficiency developed after a gastrointestinal bypass surgical procedure for morbid obesity. Five patients (39 to 60 years of age) were seen for evaluation of metabolic bone disease not responding to "usual" therapy after a gastric bypass surgical procedure. All had generalized bone pain and tenderness, muscle weakness, stooping posture, difficulty walking, and waddling gait due to severe proximal muscle weakness for a period of 2 to 5 years. Diagnoses before the referral varied from arthritis and gout to vitamin D deficiency and osteoporosis despite highly suggestive biochemical or radiologic findings (or both) of osteomalacia in each patient, which was confirmed by bone biopsy. After therapy with pharmacologic doses of ergocalciferol (100,000 IU daily) and calcium carbonate (1 to 2.5 g daily), considerable improvements occurred in clinical symptoms and functional status, biochemical indices, bone mineral density, and bone histomorphometric features. Gastric bypass operations predispose patients to severe vitamin D deficiency and osteomalacia in the absence of pharmacologic doses of vitamin D therapy. In general, the current recommendations are grossly inadequate in this high-risk population, and the clinical presentation is both nonspecific and often misleading. Prospective long-term studies are needed to determine the appropriate vitamin D dose required to prevent osteomalacia in such patients.


A novel microtubule-associated protein-2 expressed in oligodendrocytes in multiple sclerosis lesions.
Elucidation of the mechanisms involved in the regeneration of oligodendrocytes and remyelination is a central issue in multiple sclerosis (MS) research. We recently identified a novel alternatively spliced, developmentally regulated oligodendrocyte-specific protein designated microtubule-associated protein-2+13 [microtubule-associated protein-2 expressing exon 13 (MAP-2+13)]. MAP-2+13 is expressed in human fetal oligodendrocytes during process extension and myelination but is minimally expressed in normal mature CNS. To test the hypothesis that MAP-2+13 is reexpressed in regenerating oligodendrocytes in MS lesions, we examined the brains of MS patients for the expression of this protein. By immunocytochemistry using a series of monoclonal antibodies specific for MAP-2+13, we determined that MAP-2+13 expression was up-regulated in all 31 lesions from 10 different MS brains. MAP-2+13 was expressed in regenerating oligodendrocytes associated with demyelinated lesions, with the highest counts found in regions of extensive remyelination. By electron microscopy, MAP-2+13 was localized to oligodendrocytes engaged in remyelination, evident by their process extension and association with thinly myelinated (remyelinated) and demyelinated axons. These results suggest a hitherto unsuspected role for this microtubule-associated protein in oligodendrocyte function during development and myelin repair.


Resveratrol inhibits hypoxia-induced accumulation of hypoxia-inducible factor-1alpha and VEGF expression in human tongue squamous cell carcinoma and hepatoma cells.
Hypoxia-inducible factor-1alpha (HIF-1alpha) is overexpressed in many human tumors and their metastases, and is closely associated with a more aggressive tumor phenotype. In this study, we investigated the effect of resveratrol, a natural product commonly found in grapes and various other fruits, on hypoxia-induced HIF-1alpha protein accumulation and vascular endothelial growth factor (VEGF) expression in human tongue squamous cell carcinomas and hepatoma cells. Our results showed that resveratrol significantly inhibited both basal level and hypoxia-induced HIF-1alpha protein accumulation in cancer cells, but did not affect HIF-1alpha mRNA levels. Pretreatment of cells with resveratrol significantly reduced hypoxia-induced VEGF promoter activities and VEGF expression at both mRNA and protein levels. The mechanism of resveratrol inhibition of hypoxia-induced HIF-1alpha accumulation seems to involve a gradually shortened half-life of HIF-1alpha protein caused by an enhanced protein degradation through the 26S proteasome system. In addition, resveratrol remarkably inhibited hypoxia-mediated activation of extracellular signal-regulated kinase 1/2 and Akt, leading to a marked decrease in hypoxia-induced HIF-1alpha protein accumulation and VEGF transcriptional activation. Functionally, we observed that resveratrol also significantly inhibited the hypoxia-stimulated invasiveness of cancer cells. These data suggested that HIF-1alpha/VEGF could be a promising drug target for resveratrol in the development of an effective chemopreventive and anticancer therapy in human cancers.


Sports injury, occupational physical activity, joint laxity, and meniscal damage.
To investigate the risk factors for meniscal damage, an important determinant of knee osteoarthritis. We studied 243 men and women aged 20-59 years in whom the diagnosis of a meniscal tear was confirmed for the first time at arthroscopy, over a 25 month period, in 2 British hospitals. Each case was compared with one or 2 community controls, matched by age and sex, who were registered with the same general practitioner. Information on exposure to risk factors was obtained by a structured questionnaire and physical examination. Meniscal tear was strongly associated with participation in sports during the 12 months preceding the onset of symptoms; the risk was particularly high for soccer (OR 3.7; 95% CI 2.1-6.6). Higher body mass index and occupational kneeling (OR 3.8; 95% CI 1.3-11.0) and squatting (OR 2.9; 95% CI 1.0-8.0) were associated with an increased risk of degenerative meniscal lesions, after adjustment for social class, joint laxity, and sports participation. Joint laxity was associated with degenerative meniscal lesions independently of occupational physical activity, sports, and obesity. Our results confirm the importance of sporting activities entailing knee torsion in acute meniscal tear. They also point to a role for occupational activity, adiposity, and joint laxity in the pathogenesis of degenerative meniscal lesions. Modifying these mechanical risk factors may serve to reduce the risk of meniscal injury and may also help to prevent later knee osteoarthritis.


Psychological distress of cancer and clinical trial participation: a review of the literature.
The House of Commons Select Committee on Science and Technology 2000 state that currently less than 5% of adult patients with solid tumours are entered into clinical trials. They recommend that increasing the number of adult cancer patients entering clinical trials must become a high priority. Health-care providers need to prepare themselves for this proposed increase in trial participants by assessing the current status of care and implementing changes within the current infrastructure to provide optimal holistic care. Cancer can change a patient's life either for better or for worse. At one extreme, having cancer leads to enhanced appreciation of life and closer bonds with others. However, at the other extreme, cancer combined with its treatment is viewed as an event that evokes distress and emotional anguish taxing the individual's ability to cope. In the last 25 years, owing to the advent of clinical trials, progress has been made in cancer treatment. Clinical trials may be hailed as the saviour to many therapeutic dilemmas. Treatments are now available which can offer patients hope of cure. Nevertheless, many participants may fear, for the purpose of research, that they may be assigned to less than optimal therapy or that their care will be carried out in a sterile scientific atmosphere devoid of humane and personal consideration. These and other reasons may cause unacceptable personal distress that overrides the potential therapeutic gain. Cancer diagnosis coupled with the ramifications of clinical trial involvement can have significant psychological implications. They may trigger the onset of a mood disorder or exacerbate a present symptom. This article will identify mood disorders in the cancer population, focus on the participants' needs in the clinical trial arena and investigate the influence trial participation has on psychological status.


Upper respiratory tract infection in children, domestic temperatures, and humidity.
The aim of the study was to seek for a possible association between the incidence of upper respiratory tract infections and air temperature and humidity in the home. Recordings of temperature and relative humidity were made in living rooms and children's bedrooms over a six month period and related to incidence of upper respiratory tract infection. The study was carried out in one general practice of 10,000 patients. 297 children aged 24-59 months were studied, selected in random order from the practice age-sex register. Temperature and humidity recordings were made with thermohygrograph recorders over six days. Upper respiratory tract infections were recorded (a) retrospectively over the previous 12 months, and (b) during the study period. Past history of acute otitis media and recent family history of respiratory infection were also obtained. No significant association was found between the variables, although the bedrooms of children with reported upper respiratory tract infections were cooler overnight than those of non-infected children (mean difference 0.8 degrees C, 95% confidence limits 0.7 degrees C). No association was found between reported or recorded upper respiratory tract infections and age or type of home, family size, level of occupancy, social class, or smoking habits. Only 15 children (5%) were identified by their parents as having had asthma, but 58 (19.5%) had had a "wheezy chest". A greater proportion of children who wheezed slept in cooler bedrooms, had gas fires rather than central heating, and had more smokers in the house. No association between upper respiratory tract infection and domestic temperature or humidity levels could be shown in this study. Since dampness is repeatedly presented as a health risk, further study is required.


The temperature optima of enzymes: a new perspective on an old phenomenon.
Careful analysis of the dependence of enzyme activity on assay temperature has revealed that some enzymes might have real temperature optima in which the decrease in catalytic rate at temperatures above the optimum is not primarily a result of irreversible thermal inactivation. The 'equilibrium model' has been formulated to describe genuine temperature optima, and to suggest a simple experimental method by which to distinguish these cases from those in which enzyme instability is the major determinant of temperature optima.


Pharmacological Modulators of Tau Aggregation and Spreading.
Tauopathies are neurodegenerative disorders characterized by the deposition of aggregates composed of abnormal tau protein in the brain. Additionally, misfolded forms of tau can propagate from cell to cell and throughout the brain. This process is thought to lead to the templated misfolding of the native forms of tau, and thereby, to the formation of newer toxic aggregates, thereby propagating the disease. Therefore, modulation of the processes that lead to tau aggregation and spreading is of utmost importance in the fight against tauopathies. In recent years, several molecules have been developed for the modulation of tau aggregation and spreading. In this review, we discuss the processes of tau aggregation and spreading and highlight selected chemicals developed for the modulation of these processes, their usefulness, and putative mechanisms of action. Ultimately, a stronger understanding of the molecular mechanisms involved, and the properties of the substances developed to modulate them, will lead to the development of safer and better strategies for the treatment of tauopathies.


Southern African guidelines on the safe use of pre-exposure prophylaxis in persons at risk of acquiring HIV-1 infection.
The Southern African HIV Clinicians Society published its first set of oral pre-exposure prophylaxis (PrEP) guidelines in June 2012 for men who have sex with men (MSM) who are at risk of HIV infection. With the flurry of data that has been generated in PrEP clinical research since the first guideline, it became evident that there was a need to revise and expand the PrEP guidelines with new evidence of safety and efficacy of PrEP in several populations, including MSM, transgender persons, heterosexual men and women, HIV-serodiscordant couples and people who inject drugs. This need is particularly relevant following the World Health Organization (WHO) Consolidated Treatment Guidelines released in September 2015. These guidelines advise that PrEP is a highly effective, safe, biomedical option for HIV prevention that can be incorporated with other combination prevention strategies in Southern Africa, given the high prevalence of HIV in the region. PrEP should be tailored to populations at highest risk of HIV acquisition, whilst further data from studies in the region accrue to guide optimal deployment to realise the greatest impact regionally. PrEP may be used intermittently during periods of perceived HIV acquisition risk, rather than continually and lifelong, as is the case with antiretroviral treatment. Recognition and accurate measurement of potential risk in individuals and populations also warrants discussion, but are not extensively covered in these guidelines.


Clinical manifestations and coronary artery disease risk factors at diagnosis of systemic lupus erythematosus: data from an international inception cohort.
Systemic Lupus International Collaborating Clinics (SLICC) comprises 27 centres from 11 countries. An inception cohort of 918 SLE patients has been assembled according to a standardized protocol between 2000 and 2006. Clinical features, classic coronary artery disease (CAD) risk factors, as well as other potential risk factors were collected. Of the 918 patients 89% were females, and of multi racial origin. Less than half the patients were living in a permanent relationship, 58% had post secondary education and 51% were employed. Eight percent had family history of SLE. At enrolment, with at mean age of diagnosis of 34.5 years, a significant number of patients already had CAD risk factors, such as hypertension (33%) and hypercholesterolemia (36%). Only 15% of the patients were postmenopausal, 16% were current smokers and 3.6% had diabetes at entry to the SLICC-RAS (Registry for Atherosclerosis). A number of patients in this multi-racial, multi-ethnic inception cohort of lupus patients have classic CAD risk factors within a mean of 5.4 months from diagnosis. This cohort will be increased to 1500 patients to be followed yearly for 10 years. This will provide a unique opportunity to evaluate risk factors for accelerated atherosclerosis in SLE.


Relationship of some risk factors with typical and atypical manifestations of coronary heart disease.
This analysis explores whether 'typical' clinical manifestations of coronary heart disease (CHD) such as myocardial infarction and sudden death, relate to major cardiovascular risk factors in the same way as the 'atypical' manifestations, e.g. heart failure and chronic arrhythmias. Sixteen cohorts of men aged 40-59 in seven countries were examined, risk factors measured (age, systolic blood pressure, serum cholesterol and smoking habits) and 25-year mortality data collected in a systematic way. Cohorts were located in the US (n = 1), Finland (n = 2), the Netherlands (n = 1), Italy (n = 3), former Yugoslavia (n = 5), Greece (n = 2) and Japan (n = 2), with a total of 12,763 individuals. Ecological analysis based on regression equations and correlation among cohorts, and individual analyses based on proportional hazard models in pools of cohorts were conducted with typical and atypical CHD deaths as dependent variables. The ecological analysis suggests a significant relationship of populational mean levels of serum cholesterol and of systolic blood pressure to age-adjusted death rates from typical CHD manifestations. The relationships for atypical CHD deaths were not statistically significant. In the ecological approach with multivariate analysis, none of the risk factors showed relevant associations with event rates, except serum cholesterol and typical CHD deaths. The ecological relationship of serum cholesterol to atypical CHD death rates was negative but not significant. On average, mean age at death was statistically higher among atypical CHD than typical CHD patients (70.2 vs. 65.8 years). In the individual multivariate analysis conducted on pools of countries, the relationship of risk factors with typical CHD deaths was direct and significant for age, systolic blood pressure, and smoking habits in Northern Europe and America and Southern Europe, but only for systolic blood pressure and smoking habits in Japan, whereas for atypical CHD, the predictive factors were age, systolic blood pressure and cigarette smoking in Northern Europe and America and Southern Europe, but only age in Japan. The usual relationship of blood pressure and smoking habits and the differential relationship of serum cholesterol with atypical CHD (negative or absent) versus typical CHD (direct and significant) could be explained by 'two different diseases' or by a mix of poorly classified conditions among the atypical cases.


Small-for-gestational-age newborns of female refinery workers exposed to nickel.
It has not yet been established whether exposure to nickel (Ni) compounds may cause reproductive toxicity. The objective of this study was to investigate whether women employed under conditions of nickel exposure in early pregnancy were at elevated risk of delivering a newborn small-for-gestational-age (SGA). A register-based study of a well defined population. Data on pregnancy outcome and maternal occupation were obtained from the Kola Birth Registry. Each birth record was assigned a Ni exposure rating category according to maternal occupation at the time of becoming pregnant. Nickel exposure assessment was based on determining the water-soluble Ni subfraction of respirable aerosol fraction obtained by personal monitoring, and/or on measurements of urine Ni concentration. The reference population were the delivering women with background exposure level. The study population consisted of 22 836 births (>27 weeks of gestation) and the SGA infants were defined as below the 10th percentile birth weight for gestational age in the source population. Multiple logistic regression was used to analyze the association of the outcome with the assigned exposure rating category. The adjusted odds ratio for Ni-exposed women for giving birth to an SGA newborn was 0.84 (95% CI: 0.75-0.93). We found no adverse effect of maternal occupational exposure to water-soluble Ni in the first part of pregnancy on the risk of delivering an SGA newborn without trisomy. The finding does not exclude a possibility that exposure throughout pregnancy might produce such an effect.


Lesions of the human amygdala impair enhanced perception of emotionally salient events.
Commensurate with the importance of rapidly and efficiently evaluating motivationally significant stimuli, humans are probably endowed with distinct faculties and maintain specialized neural structures to enhance their detection. Here we consider that a critical function of the human amygdala is to enhance the perception of stimuli that have emotional significance. Under conditions of limited attention for normal perceptual awareness-that is, the attentional blink-we show that healthy observers demonstrate robust benefits for the perception of verbal stimuli of aversive content compared with stimuli of neutral content. In contrast, a patient with bilateral amygdala damage has no enhanced perception for such aversive stimulus events. Examination of patients with either left or right amygdala resections shows that the enhanced perception of aversive words depends specifically on the left amygdala. All patients comprehend normally the affective meaning of the stimulus events, despite the lack of evidence for enhanced perceptual encoding of these events in patients with left amygdala lesions. Our results reveal a neural substrate for affective influences on perception, indicating that similar neural mechanisms may underlie the affective modulation of both recollective and perceptual experience.


Kappa-opioid potentiation of tumor necrosis factor-alpha-induced anti-HIV-1 activity in acutely infected human brain cell cultures.
Opioids have been postulated to play an immunomodulatory role in the pathogenesis of HIV-1. Synthetic kappa-opioid receptor (KOR) ligands have been found to inhibit HIV-1 expression in acutely infected microglial cell cultures. We recently found that interleukin(IL)-1beta and tumor necrosis factor(TNF)-alpha have antiviral effects in acutely infected mixed glial/neuronal cell cultures. In the present study, we investigated whether selective KOR ligands would exert antiviral effects in acutely infected brain cell cultures. While the KOR ligand trans-3,4-dichloro-N-methyl-N[2-(1-pyrolidinyl)cyclohexyl]benze neaceamide methanesulfonate (U50,488) alone had little anti-HIV-1 activity, this opioid potentiated in a concentration-dependent manner the antiviral activity of TNF-alpha, but not of IL-1beta. The potentiating effect of U50,488 was detected after a 6-hr pretreatment and peaked at 24 hr. The KOR antagonist nor-binaltorphimine completely blocked the potentiating effect of U50,488, suggesting the involvement of a KOR-mediated mechanism. Antibodies to TNF-alpha completely blocked the potentiating effect of U50,488, suggesting a critical role for TNF-alpha. Antibodies to IL-1beta blocked the potentiating effect of U50,488, suggesting that IL-1beta was released following U50,488 treatment, which might contribute to the potentiating effect of U50,488. These in vitro findings support the notion that synthetic kappa-opioids could be considered as potential adjunctive therapeutic agents in HIV-1-related brain disease.


EMEA and the evaluation of health-related quality of life data in the drug regulatory process.
This study reviewed the European guidelines of the Committee for Proprietary Medicinal Products (CPMP) on how health-related quality of life (HRQOL) research should be conducted in clinical trials. Published product-level information was also reviewed to investigate the actual role of HRQOL data in the European regulatory process. All disease-specific notes for guidance and concept papers on clinical investigations, development and evaluation of human medicinal products, as well as the European Public Assessment Reports (EPAR) of all approved drugs published on the European Agency for the Evaluation of Medicinal Products (EMEA) Web site were evaluated for their HRQOL recommendations. Only twenty of the fifty CPMP guidance notes for clinical investigation of pharmaceutical products in specific disease areas included a reference to HRQOL. Most of the recommendations were generic and vague, and the terminology used was inconsistent across documents. The EPAR provided nonspecific information about HRQOL and contradictory conclusions on the effect of a drug on HRQOL sometimes occurred in different documents. The criteria used by the CPMP to assess the HRQOL data could not be identified due to an ad hoc approach to the inclusion of data in the EPAR. A more systematic approach is needed on the way health outcomes data are considered, reviewed, and interpreted by the regulatory authorities. For this to be achieved, CPMP should develop general guidelines on the importance of HRQOL and how research should be conducted if data are to be included in the registration process.


Enhancement of graphene visibility on transparent substrates by refractive index optimization.
Optical reflection microscopy is one of the main imaging tools to visualize graphene microstructures. Here is reported a novel method that employs refractive index optimization in an optical reflection microscope, which greatly improves the visibility of graphene flakes. To this end, an immersion liquid with a refractive index that is close to that of the glass support is used in-between the microscope lens and the support improving the contrast and resolution of the sample image. Results show that the contrast of single and few layer graphene crystals and structures can be enhanced by a factor of 4 compared to values commonly achieved with transparent substrates using optical reflection microscopy lacking refractive index optimization.


Immunopeptidomic analyses of colorectal cancers with and without microsatellite instability.
Colorectal cancer is the second leading cause of cancer death worldwide, and the incidence of this disease is expected to increase as global socioeconomic changes occur. Immune checkpoint inhibition therapy is effective in treating a minority of colorectal cancer tumors; however, microsatellite stable tumors do not respond well to this treatment. Emerging cancer immunotherapeutic strategies aim to activate a cytotoxic T cell response against tumor-specific antigens, presented exclusively at the cell surface of cancer cells. These antigens are rare and are most effectively identified with a mass spectrometry-based approach, which allows the direct sampling and sequencing of these peptides. Although the few tumor-specific antigens identified to date are derived from coding regions of the genome, recent findings indicate that a large proportion of tumor-specific antigens originate from allegedly noncoding regions. Here, we employed a novel proteogenomic approach to identify tumor antigens in a collection of colorectal cancer-derived cell lines and biopsy samples consisting of matched tumor and normal adjacent tissue. The generation of personalized cancer databases paired with mass spectrometry analyses permitted the identification of more than 30,000 unique MHC I-associated peptides. We identified 19 tumor-specific antigens in both microsatellite stable and unstable tumors, over two-thirds of which were derived from noncoding regions. Many of these peptides were derived from source genes known to be involved in colorectal cancer progression, suggesting that antigens from these genes could have therapeutic potential in a wide range of tumors. These findings could benefit the development of T cell-based vaccines, in which T cells are primed against these antigens to target and eradicate tumors. Such a vaccine could be used in tandem with existing immune checkpoint inhibition therapies, to bridge the gap in treatment efficacy across subtypes of colorectal cancer with varying prognoses. Data are available via ProteomeXchange with identifier PXD028309.


When to conduct probabilistic linkage vs. deterministic linkage? A simulation study.
When unique identifiers are unavailable, successful record linkage depends greatly on data quality and types of variables available. While probabilistic linkage theoretically captures more true matches than deterministic linkage by allowing imperfection in identifiers, studies have shown inconclusive results likely due to variations in data quality, implementation of linkage methodology and validation method. The simulation study aimed to understand data characteristics that affect the performance of probabilistic vs. deterministic linkage. We created ninety-six scenarios that represent real-life situations using non-unique identifiers. We systematically introduced a range of discriminative power, rate of missing and error, and file size to increase linkage patterns and difficulties. We assessed the performance difference of linkage methods using standard validity measures and computation time. Across scenarios, deterministic linkage showed advantage in PPV while probabilistic linkage showed advantage in sensitivity. Probabilistic linkage uniformly outperformed deterministic linkage as the former generated linkages with better trade-off between sensitivity and PPV regardless of data quality. However, with low rate of missing and error in data, deterministic linkage performed not significantly worse. The implementation of deterministic linkage in SAS took less than 1min, and probabilistic linkage took 2min to 2h depending on file size. Our simulation study demonstrated that the intrinsic rate of missing and error of linkage variables was key to choosing between linkage methods. In general, probabilistic linkage was a better choice, but for exceptionally good quality data (<5% error), deterministic linkage was a more resource efficient choice.


Independent and interactive effects of apolipoprotein E phenotype and cardiorespiratory fitness on plasma lipids.
To examine whether the association Apolipoprotein E (Apo E) phenotype with plasma lipids is influenced by physical fitness level. Also, to explore the interactive and independent relative contributions of Apo E phenotype, fitness (or physical activity), and other modifiable factors to variation in plasma low density lipoprotein (LDL-C) and high density lipoprotein (HDL-C) levels at baseline and over a seven-year follow-up. Physical fitness (duration of a graded treadmill test), Apo E phenotype, plasma LDL-C and HDL-C, and covariates were measured at baseline and seven years later in a bi-racial cohort of young adults, aged 18-30 years at baseline in 1985-86, from the Coronary Artery Risk Development In Young Adults (CARDIA) study. Fitness did not influence the associations of Apo E and LDL-C or HDL-C. The independent effects of several modifiable variables (changes in Keys' score, smoking, oral contraceptive use, education, body weight, alcohol intake, and fitness), when combined, contributed considerably more than Apo E to the variance in LDL-C changes (6.74% or 8.71% for combined modifiable variables vs. 1.27% or 0.90% for ApoE, in women or men, respectively) and HDL-C changes (13.11% or 12.66% for combined modifiable variables vs. 0.12% or 0.02% for ApoE, in women or men, respectively). The pattern of findings was similar when self-reported physical activity was substituted for fitness. Changes in modifiable factors, including fitness, may be stronger correlates of changes in LDL-C and HDL-C over time than the immutable factor, Apo E phenotype.


Long-term effects of brief substance use interventions for mandated college students: sleeper effects of an in-person personal feedback intervention.
It is known that brief interventions for mandated college students decrease alcohol use and/or related problems in the short term. However, none of the existing studies has followed students' past 6 months. Therefore, we compared the long-term efficacy of 2 brief substance use feedback interventions for mandated college students. We followed up mandated students (N=348) who were randomly assigned to either a brief motivational interview (BMI; n=180) or a written feedback-only (WF; n=168) intervention at 4 months and 15 months postintervention. Long-term follow-up data revealed that students, at the aggregate level, decreased their peak blood alcohol concentration (BAC) levels, number of drinks per week, and number of alcohol-related problems at 15 months postintervention compared with their baseline levels. With the exception of peak BAC, the observed long-term reduction was mainly due to the positive change among students who received the BMI intervention. Students in the BMI intervention showed significantly lower levels of alcohol-related problems at 15 months than those in the WF intervention. The BMI intervention more effectively reduced within-individual alcohol-related problems during the initial 4 months, and more successfully curbed the subsequent increase in alcohol use frequency and number of drinks per week during the 11 months between the 2 follow-up assessments. The results suggest that brief substance use interventions reduce the riskiest type of alcohol use (e.g., peak BAC) among mandated college students over the long term, and that sleeper effects of in-person personal feedback interventions (PFIs) exist. In-person PFIs in the context of a motivational interview may be more efficacious in the long term than written feedback-only interventions for mandated students. Future studies comparing interventions for college students should extend follow-up for longer periods of time.


Purinergic receptors in the brainstem mediate hypotension and bradycardia.
Adenosine acts at many sites to modulate neuronal activity. The purpose of this study was to investigate a possible role for adenosine as a neuromodulator of brainstem cardiovascular control. Microinjections of adenosine (0-2.3 nmol) were made stereotaxically into various brainstem sites. Injection of adenosine into the nucleus tractus solitarii (NTS) produced dose-related decreases in heart rate and systolic and diastolic blood pressures. Maximal changes occurred 90 seconds after injection. Injection into the area postrema also produced decreased heart rate and systolic and diastolic blood pressures. No significant effect occurred following injection into the C1 area. Adenosine 5'-triphosphate and its analogue, beta, gamma-methylene adenosine 5'-triphosphate also produced dose-related and potent vasodepressor and bradycardia effects in the NTS. Injection of 1,3-dipropyl-8-p-sulfophenylxanthine (0.92 nmol), a potent adenosine receptor antagonist, produced no effect itself, but abolished for 45 minutes the actions of further injections of adenosine and adenosine 5'-triphosphate (but not L-glutamate) in both the NTS and area postrema. Thus, NTS and area postrema injections of adenosine decrease blood pressure and heart rate in anesthetized normotensive rats through adenosine receptors located in these areas. These findings support a role for endogenous adenosine as a central modulator in cardiovascular control.


An assessment of quality of home-based HIV counseling and testing performed by lay counselors in a rural sub-district of KwaZulu-Natal, South Africa.
HIV counseling and testing (HCT) has been prioritized as one of the prevention strategies for HIV/AIDS, and promoted as an essential tool in scaling up and improving access to treatment, care and support especially in community settings. Home-based HCT (HBHCT) is a model that has consistently been found to be highly acceptable and has improved HCT coverage and uptake in low- and middle-income countries since 2002. It involves trained lay counselors going door-to-door offering pre-test counseling and providing HCT services to consenting eligible household members. Currently, there are few studies reporting on the quality of HBHCT services offered by lay counselors especially in Sub-Saharan Africa, including South Africa. This is a quantitative descriptive sub-study of a community randomized trial (Good Start HBHCT trial) which describes the quality of HBHCT provided by lay counselors. Quality of HBHCT was measured as scores comparing observed practice to prescribed protocols using direct observation. Data were collected through periodic observations of HCT sessions and exit interviews with clients. Counselor quality scores for pre-test counseling and post-test counseling sessions were created to determine the level of quality. For the client exit interviews a continuous score was created to assess how satisfied the clients were with the counseling session. A total of 196 (3%) observational assessments and 406 (6%) client exit interviews were completed. Overall, median scores for quality of counseling and testing were high for both HIV-negative and HIV-positive clients. For exit interviews all 406 (100%) clients had overall satisfaction with the counseling and testing services they received, however 11% were concerned about the counselor keeping their discussion confidential. Of all 406 clients, 393 (96.8%) intended to recommend the service to other people. In ensuring good quality HCT services, ongoing quality assessments are important to monitor quality of HCT after training.


[Investigation of herpes simplex virus in viral meningoencephalitis suspected cases using molecular and serological methods].
Herpes simplex virus (HSV) meningoencephalitis has a high mortality rate if proper antiviral therapy is not applied. Thus rapid diagnosis is of peculiar importance in such cases. In this study we aimed to evaluate the use of polymerase chain reaction (PCR) and detection of intrathecally synthesized antibodies by serological methods in viral meningoencephalitis suspected cases to determine HSV as the causative agent. Seventeen cases with cerebrospinal fluid (CSF) samples with microscopical and biochemical findings compatible with viral encephalitis were included in this study. CSF samples yielded no bacterial growth. Cell cultures propagated in Vero cell line and PCR with different primer sets against HSV-1 and HSV-2 (specific for US7 and US2 gene regions, respectively) were used to investigate the presence of HSV in CSF. Serum samples were taken simultaneously with the CSF sampling and "Reibergram" graphics and antibody index (AI) calculations were used for the evaluation of intrathecal antibody synthesis. Albumin and total IgG levels in serum and CSF samples measured with nephelometry (Dade-Behring, Germany) for Reibergram graphics, albumin and IgG ratios were calculated. Quantitative levels of HSV 1+2 IgG were measured in serum and CSF samples using ELISA (HSV-1/2 Pool; Antibody Determination in CSF, Euroimmun, Germany) for AI determination. One of the CSF samples obtained one week after the neurological symptoms had started, yielded HSV-2 in cell culture and also HSV-2 DNA was detected by PCR. Intrathecal antibody synthesis was not detected in this case. In two cases with symptoms lasting for more than three weeks, intrathecal IgG synthesis in Reibergrams and pathological intrathecal HSV AI (27.9 and 7.9, respectively) were detected, however, virus isolation and PCR detection were not successful. For the other 14 cases, HSV-DNA were found negative and no intrathecal antibody synthesis were detected. HSV meningoencephalitis can be diagnosed via using PCR which has been accepted as gold standard in recent years, with 24 hours turn-around time. However, if CSF samples were taken later than the first week following the beginning of symptoms, possibility of HSV detection by PCR is lowered. According to the data obtained from this limited study, it may be suggested that PCR may be used for the detection of HSV-DNA in CSF samples during the early phase of meningoencephalitis cases, however, consideration must be taken to detect the intrathecal antibodies during the later phases of the infection where intrathecal antibody synthesis starts.


RhoB is dispensable for mouse development, but it modifies susceptibility to tumor formation as well as cell adhesion and growth factor signaling in transformed cells.
RhoB is an endosomal small GTPase that is implicated in the response to growth factors, genotoxic stress, and farnesyltransferase inhibitors. To gain insight into its physiological functions we examined the consequences of homozygous gene deletion in the mouse. Loss of RhoB did not adversely affect mouse development, fertility, or wound healing. However, embryo fibroblasts cultured in vitro exhibited a defect in motility, suggesting that RhoB has a role in this process that is conditional on cell stress. Neoplastic transformation by adenovirus E1A and mutant Ras yielded differences in cell attachment and spreading that were not apparent in primary cells. In addition, transformed -/- cells displayed altered actin and proliferative responses to transforming growth factor beta. A negative modifier role in transformation was suggested by the increased susceptibility of -/- mice to 7,12-dimethylbenz[a]anthracene-induced skin carcinogenesis and by the increased efficiency of intraperitoneal tumor formation by -/- cells. Our findings suggest that RhoB is a negative regulator of integrin and growth factor signals that are involved in neoplastic transformation and possibly other stress or disease states.


Subpopulations of cholinergic, GABAergic and glutamatergic neurons in the pedunculopontine nucleus contain calcium-binding proteins and are heterogeneously distributed.
Neurons in the pedunculopontine nucleus (PPN) are highly heterogeneous in their discharge properties, their neurochemical markers, their pattern of connectivity and the behavioural processes in which they participate. Three main transmitter phenotypes have been described, cholinergic, GABAergic and glutamatergic, and yet electrophysiological evidence suggests heterogeneity within these subtypes. To gain further insight into the molecular composition of these three populations in the rat, we investigated the pattern of expression of calcium binding proteins (CBPs) across distinct regions of the PPN and in relation to the presence of other neurochemical markers. Calbindin- and calretinin-positive neurons are as abundant as cholinergic neurons, and their expression follows a rostro-caudal gradient, whereas parvalbumin is expressed by a low number of neurons. We observed a high degree of expression of CBPs by GABAergic and glutamatergic neurons, with a large majority of calbindin- and calretinin-positive neurons expressing GAD or VGluT2 mRNA. Notably, CBP-positive neurons expressing GAD mRNA were more concentrated in the rostral PPN, whereas the caudal PPN was characterized by a higher density of CBP-positive neurons expressing VGluT2 mRNA. In contrast to these two large populations, in cholinergic neurons expression of calretinin is observed only in low numbers and expression of calbindin is virtually non-existent. These findings thus identify novel subtypes of cholinergic, GABAergic and glutamatergic neurons based on their expression of CBPs, and further contribute to the notion of the PPN as a highly heterogeneous structure, an attribute that is likely to underlie its functional complexity.


Hip Capsular Management in Patients With Femoroacetabular Impingement or Microinstability: A Systematic Review of Biomechanical Studies.
To investigate the correlation between hip capsular management (repair or reconstruction) and biomechanical results in the setting of femoroacetabular impingement and microinstability. A search of the PubMed and Embase databases was performed using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Included studies focused on hip biomechanics related to capsular release, repair of I- and T-capsulotomy, or capsular reconstruction. Studies were assessed for external/internal rotation of the femur, femoral head translation, rotational torque, and distraction force. Articles were excluded if they discussed treatment of the hip capsule related to surgical dislocation, mini-open surgery, arthroplasty, reorientation osteotomy, or traumatic dislocation. Twenty-four biomechanical studies were included that evaluated rotation/translation (11 studies), distraction (3 studies), the capsular role in microinstability (simulated with anterior capsule pie crusting [2 studies] and cyclical loading [2 studies]), allograft reconstruction (3 studies), and anatomic properties (3 studies). Repair and reconstruction demonstrated improvements in maximum distractive force, total ROM, and torsional stability when compared to capsular release. Significant differences were observed between capsular repair and release in total ROM in the coronal plane with improved stability in the repair groups (standardized mean difference [SMD]: -1.3°, 95% confidence interval [CI] -1.68 , -0.854; P < .001). There was significantly increased total motion in the coronal plane in the capsular laxity state compared to the native state (SMD: 1.4° (95% CI 0.32, 2.49; P = .012). Biomechanical evidence supports closure of the capsule after hip arthroscopy to reverse the significant effects of capsulotomy. Simulated capsule laxity models created altered joint motion and translation. Capsule reconstruction appears to restore the hip to its native capsule state. Investigating the biomechanical outcomes of capsular repair and reconstruction will help surgeons better understand the rationale and implications of these capsular management strategies.


Particle Physics in High School: A Diagnose Study.
The science learning process improves when the contents are connected to students' lives. Particle physics has had a great impact in our society in the last years and has changed the theoretical picture about matter fundamental dynamics. Thus, we think that academic contents about matter components and interactions should be updated. With this study we aim to characterize the level of knowledge of high school students about this topic. We built a test with questions about classical atomic models, particle physics, recent discoveries, social implications and students opinions about it. Contrary to our first suspicion, students' answers show a high variability. They have new physics ideas and show a great interest towards modern concepts. We suggest including an updated view of this topic as part of the curriculum.


A LAD-III syndrome is associated with defective expression of the Rap-1 activator CalDAG-GEFI in lymphocytes, neutrophils, and platelets.
Leukocyte and platelet integrins rapidly alter their affinity and adhesiveness in response to various activation (inside-out) signals. A rare leukocyte adhesion deficiency (LAD), LAD-III, is associated with severe defects in leukocyte and platelet integrin activation. We report two new LAD cases in which lymphocytes, neutrophils, and platelets share severe defects in beta(1), beta(2), and beta(3) integrin activation. Patients were both homozygous for a splice junction mutation in their CalDAG-GEFI gene, which is a key Rap-1/2 guanine exchange factor (GEF). Both mRNA and protein levels of the GEF were diminished in LAD lymphocytes, neutrophils, and platelets. Consequently, LAD-III platelets failed to aggregate because of an impaired alpha(IIb)beta(3) activation by key agonists. beta(2) integrins on LAD-III neutrophils were unable to mediate leukocyte arrest on TNFalpha-stimulated endothelium, despite normal selectin-mediated rolling. In situ subsecond activation of neutrophil beta(2) integrin adhesiveness by surface-bound chemoattractants and of primary T lymphocyte LFA-1 by the CXCL12 chemokine was abolished. Chemokine inside-out signals also failed to stimulate lymphocyte LFA-1 extension and high affinity epitopes. Chemokine-triggered VLA-4 adhesiveness in T lymphocytes was partially defective as well. These studies identify CalDAG-GEFI as a critical regulator of inside-out integrin activation in human T lymphocytes, neutrophils, and platelets.


Stem cell characterization of neutropenia: velocity sedimentation and mass culture analysis.
Human bone marrow obtained from patients with neutropenia contains a cell population which is absent or diminished in normal marrow. The abnormal population is composed of cells of volume 200-300 mum3 which sediment at 5-5 to 8-5 mm/h. Normal marrow contains one cell class giving rise to increased numbers of grnaulocyte colonies after mass culture, while marrow obtained from neutropenic patients, or from patients with marrow regeneration, shows two such populations; one of these cell classes corresponds to the abnormally large cells demonstrated on velocity sedimentation analysis. This population of large cells may represent a group of either self-renewing cells related to the committed granulocyte progenitors or the pluripotent stem cell.


Mechanisms of vascular dysfunction in mice with endothelium-specific deletion of the PPAR-δ gene.
Peroxisome proliferator-activated receptor (PPAR)-δ is a nuclear hormone receptor that is mainly involved in lipid metabolism. Recent studies have suggested that PPAR-δ agonists exert vascular protective effects. The present study was designed to characterize vascular function in mice with genetic inactivation of PPAR-δ in the endothelium. Mice with vascular endothelial cell-specific deletion of the PPAR-δ gene (ePPARδ(-/-) mice) were generated using loxP/Cre technology. ePPARδ(-/-) mice were normotensive and did not display any sign of metabolic syndrome. Endothelium-dependent relaxations to ACh and endothelium-independent relaxations to the nitric oxide (NO) donor diethylammonium (Z)-1-(N,N-diethylamino)diazen-1-ium-1,2-diolate were both significantly impaired in the aorta and carotid arteries of ePPARδ(-/-) mice (P < 0.05). In ePPARδ(-/-) mouse aortas, phosphorylation of endothelial NO synthase at Ser(1177) was significantly decreased (P < 0.05). However, basal levels of cGMP were unexpectedly increased (P < 0.05). Enzymatic activity of GTP-cyclohydrolase I and tetrahydrobiopterin levels were also enhanced in ePPARδ(-/-) mice (P < 0.05). Most notably, endothelium-specific deletion of the PPAR-δ gene significantly decreased protein expressions of catalase and glutathione peroxidase 1 and resulted in increased levels of H2O2 in the aorta (P < 0.05). In contrast, superoxide anion production was unaltered. Moreover, treatment with catalase prevented the endothelial dysfunction and elevation of cGMP detected in aortas of ePPARδ(-/-) mice. The findings suggest that increased levels of cGMP caused by H2O2 impair vasodilator reactivity to endogenous and exogenous NO. We speculate that chronic elevation of H2O2 predisposes PPAR-δ-deficient arteries to oxidative stress and vascular dysfunction.


Nondestructive method for measuring the degree of cluster-induced quenching in er(3+)-doped waveguides and fibers.
Ion clustering is one of the major dissipative processes in the operation of Er(3+)-doped planar waveguide amplifiers and heavily doped fiber amplifiers. We propose and demonstrate a nondestructive method for measuring the fraction of erbium ions in doped planar waveguides and fibers that are quenched by ultrafast parasitic energy transfer in ion pairs or clusters. The method is based on unsaturable absorption measurements in the 980-nm absorption band. By combining transmission data at resonance with the corresponding data for radiation outside the absorption band, the influence of background and coupling losses can be eliminated.


The spatial distribution of radiodense breast tissue: a longitudinal study.
Mammographic breast density is one of the strongest known markers of susceptibility to breast cancer. To date research into density has relied on a single measure (for example, percent density (PD)) summarising the average level of density for the whole breast, with no consideration of how the radiodense tissue may be distributed. This study aims to investigate the spatial distribution of density within the breast using 493 mammographic images from a sample of 165 premenopausal women (~3 medio-lateral oblique views per woman). Each breast image was divided into 48 regions and the PD for the whole breast (overall PD) and for each one of its regions (regional PD) was estimated. The spatial autocorrelation (Moran's I value) of regional PD for each image was calculated to investigate spatial clustering of density, whether the degree of clustering varied between a woman's two breasts and whether it was affected by age and other known density correlates. The median Moran's I value for 165 women was 0.31 (interquartile range: 0.26, 0.37), indicating a clustered pattern. High-density areas tended to cluster in the central regions of the breast, regardless of the level of overall PD, but with considerable between-woman variability in regional PD. The degree of clustering was similar between a woman's two breasts (mean within-woman difference in Moran's I values between left and right breasts = 0.00 (95% confidence interval (CI) = -0.01, 0.01); P = 0.76) and did not change with aging (mean within-woman difference in I values between screens taken on average 8 years apart = 0.01 (95% CI = -0.01, 0.02); P = 0.30). Neither parity nor age at first birth affected the level of spatial autocorrelation of density, but increasing body mass index (BMI) was associated with a decrease in the degree of spatial clustering. This study is the first to demonstrate that the distribution of radiodense tissue within the breast is spatially autocorrelated, generally with the high-density areas clustering in the central regions of the breast. The degree of clustering was similar within a woman's two breasts and between women, and was little affected by age or reproductive factors although it declined with increasing BMI.


Workplace Measurements of Ultrafine Particles-A Literature Review.
Workers are exposed to ultrafine particles (UFP) in a number of occupations. In order to summarize the current knowledge regarding occupational exposure to UFP (excluding engineered nanoparticles), we gathered information on UFP concentrations from published research articles. The aim of our study was to create a basis for future epidemiological studies that treat UFP as an exposure factor. The literature search found 72 publications regarding UFP measurements in work environments. These articles covered 314 measurement results and tabled concentrations. Mean concentrations were compared to typical urban UFP concentration level, which was considered non-occupational background concentration. Mean concentrations higher than the typical urban UFP concentration were reported in 240 workplace measurements. The results showed that workers' exposure to UFP may be significantly higher than their non-occupational exposure to background concentration alone. Mean concentrations of over 100 times the typical urban UFP concentration were reported in welding and metal industry. However, according to the results of the review, measurements of the UFP in work environments are, to date, too limited and reported too heterogeneous to allow us to draw general conclusions about workers' exposure. Harmonization of measurement strategies is essential if we are to generate more reliable and comparable data in the future.


Chondrogenic Differentiation of Human Mesenchymal Stem Cells Results in Substantial Changes of Ecto-Nucleotides Metabolism.
Mesenchymal stem cells (MSCs) are population of adult stem cells and attractive candidates for cartilage repair due to their chondrogenic potential. Purinergic compounds (purinergic receptors and ecto-enzymes metabolizing nucleotides), together with nucleotides/nucleosides present in the extracellular environment, are known to play a key role in controlling the stem cells biological potential to proliferate and differentiate. Despite the available literature pointing to the importance of purinergic signaling in controlling the fate of MSCs, the research results linking nucleotides and ecto-nucleotidases with MSCs chondrogenic differentiation are indigent. Therefore, the aim of presented study was the characterization of the ecto-nucleotides hydrolysis profile and ecto-enzymes expression in human umbilical cord-derived MSCs and chondrogenically induced MSCs. We described substantial changes of ecto-nucleotides metabolism and ecto-enzymes expression profiles resulting from chondrogenic differentiation of human umbilical cord-derived MSCs. The increased rate of ADP hydrolysis, measured by ecto-nucleotidases activity, plays a pivotal role in the regulation of cartilage formation and resorption. Despite the increased level of NTPDase1 and NTPDase3 mRNA expression in chondrogenically induced MSCs, their activity toward ATP remains quite low. Supported by the literature data, we hypothesize that structure-function relationships in chondrogenic lineage dictate the direction of nucleotides metabolism. In early neocartilage tissue, the beneficial role of ATP in improving biomechanical properties of cartilage does not necessitate the high rate of enzymatic ATP degradation.


Identification of the alternatively spliced exons of murine CD45 (T200) required for reactivity with B220 and other T200-restricted antibodies.
Cell lines expressing specific variants of murine CD45 (T200) were established by infection with retroviral constructs of four cDNAs encoding different forms of CD45. These lines were then used to determine which sequences encoded by alternatively spliced exons of CD45 were required to generate antigenic determinants recognized by anti-CD45 mAbs. The binding of two B220 antibodies (14.8 and RA32C2) to CD45 was dependent on the expression of the first alternatively spliced exon (exon A). A third B220 antibody, RA3-6B2, did not bind to any of the forms of CD45 expressed on fibroblasts. A newly defined anti-CD45R antibody, C363.16A, reacts with an antigenic site dependent upon the expression of the second alternatively spliced exon, exon B.


SAXS Data Alone can Generate High-Quality Models of Protein-Protein Complexes.
Modeling driven by small-angle X-ray scattering (SAXS) combines low-resolution data with computational modeling to predict the structure of biomolecular assemblies. A new protocol, ATTRACT-SAXS, has been developed and tested on a large protein-protein docking benchmark with simulated SAXS data. For 88% of cases, high-quality solutions were generated using SAXS data alone without a physiochemical force field (interface-RMSD ≦ 2 Å or ligand-RMSD ≦ 5 Å; and more than 30% native contacts). ATTRACT-SAXS gave significant improvements compared with previous approaches that filter by SAXS a posteriori. When combining SAXS and interface properties for scoring, the protocol placed high-quality models in 70% of cases among the top-ranked 100 clusters. ATTRACT-SAXS also gave good results when tested on experimental data if the native complex structure was compatible with the SAXS profile. Our results show that, in principle, SAXS on its own can contain enough information to generate high-quality models of protein-protein complexes.


Hospital-Acquired Methicillin-Resistant Staphylococcus aureus Bloodstream Infections in Québec: Impact of Guidelines.
OBJECTIVE We examined the impact of methicillin-resistant Staphylococcus aureus (MRSA) guidelines in Québec adult hospitals from January 1, 2006, to March 31, 2015, by examining the incidence rate reduction (IRR) in healthcare-associated MRSA bloodstream infections (HA-MRSA), using central-line associated bloodstream infections (CLABSIs) as a comparator. METHODS In this study, we utilized a quasi-experimental design with Poisson segmented regression to model HA-MRSA and CLABSI incidence for successive 4-week surveillance segments, stratified by teaching status. We used 3 distinct periods with 2 break points (April 1, 2007, and January 3, 2010) corresponding to major MRSA guideline publications and updates. RESULTS Over the study period, HA-MRSA incidence decreased significantly in adult teaching facilities but not in nonteaching facilities. Prior to MRSA guideline publication (2006-2007), HA-MRSA incidence decrease was not significant (P=.89), while CLABSI incidence decreased by 4% per 4-week period (P=.05). After the publication of guidelines (2007-2009), HA-MRSA incidence decreased significantly by 1% (P=.04), while no significant decrease in CLABSI incidence was observed (P=.75). HA-MRSA and CLABSI decreases were both significant at 1% for 2010-2015 (P<.001 and P=.01, respectively). These decreases were gradual rather than sudden; break points were not significant. Teaching facilities drove these decreases. CONCLUSION During the study period, HA-MRSA and CLABSI rates decreased significantly. In 2007-2009, the significant decrease in HA-MRSA rates with stable CLABSI rates suggests an impact from MRSA-specific guidelines. In 2010-2015, significant and equal IRRs for HA-MRSA and CLABSI may be due to the continuing impact of MRSA guidelines, to the impact of new interventions targeting device-associated infections in general by the 2010-2015 Action Plan, or to a combination of factors. Infect Control Hosp Epidemiol 2017;38:840-847.


A cis-regulatory map of the Drosophila genome.
Systematic annotation of gene regulatory elements is a major challenge in genome science. Direct mapping of chromatin modification marks and transcriptional factor binding sites genome-wide has successfully identified specific subtypes of regulatory elements. In Drosophila several pioneering studies have provided genome-wide identification of Polycomb response elements, chromatin states, transcription factor binding sites, RNA polymerase II regulation and insulator elements; however, comprehensive annotation of the regulatory genome remains a significant challenge. Here we describe results from the modENCODE cis-regulatory annotation project. We produced a map of the Drosophila melanogaster regulatory genome on the basis of more than 300 chromatin immunoprecipitation data sets for eight chromatin features, five histone deacetylases and thirty-eight site-specific transcription factors at different stages of development. Using these data we inferred more than 20,000 candidate regulatory elements and validated a subset of predictions for promoters, enhancers and insulators in vivo. We identified also nearly 2,000 genomic regions of dense transcription factor binding associated with chromatin activity and accessibility. We discovered hundreds of new transcription factor co-binding relationships and defined a transcription factor network with over 800 potential regulatory relationships.


Late versus early repair of Achilles tendon rupture. Clinical and biomechanical evaluation.
Eleven patients who had late reconstruction for Achilles tendon rupture were compared with 10 patients who had immediate repair. The followup period averaged 8 years for both groups. Results, based on a clinical score, were successful (excellent or good) for all but 1 patient. Although manual testing demonstrated normal plantar flexion strength in all but 1 patient, significant differences were seen in isometric and isokinetic plantar flexion strength of the involved sides as compared with the uninvolved sides of these patients. The patient with the unsuccessful result had abnormalities in vertical, foreaft, and medial-lateral force components of the ground reaction force. Three-dimensional motion analysis showed decreased total motion in the sagittal plane on the operated side as compared with the unoperated side in this patient. The data indicated that patients with late reconstruction of Achilles tendon rupture have successful clinical results comparable with those of early repair.


Voltage and Deflection Amplification via Double Resonance Excitation in a Cantilever Microstructure.
Cantilever electrostatically-actuated resonators show great promise in sensing and actuating applications. However, the electrostatic actuation suffers from high-voltage actuation requirements and high noise low-amplitude signal-outputs which limit its applications. Here, we introduce a mixed-frequency signal for a cantilever-based resonator that triggers its mechanical and electrical resonances simultaneously, to overcome these limitations. A single linear RLC circuit cannot completely capture the response of the resonator under double resonance excitation. Therefore, we develop a coupled mechanical and electrical mathematical linearized model at different operation frequencies and validate this model experimentally. The double-resonance excitation results in a 21 times amplification of the voltage across the resonator and 31 times amplitude amplification over classical excitation schemes. This intensive experimental study showed a great potential of double resonance excitation providing a high amplitude amplification and maintaining the linearity of the system when the parasitic capacitance is maintained low.


Identification of space curves from two-dimensional perspective views.
This paper describes a new method to be used for matching three-dimensional objects with curved surfaces to two-dimensional perspective views. The method requires for each three-dimensional object a stored model consisting of a closed space curve representing some characteristic connected curved edges of the object. The input is a two-dimensional perspective projection of one of the stored models represented by an ordered sequence of points. The input is converted to a spline representation which is sampled at equal intervals to derive a curvature function. The Fourier transform of the curvature function is used to represent the shape. The actual matching is reduced to a minimization problem which is handled by the Levenberg-Marquardt algorithm [3].


What have we learned from recent twin studies about the etiology of neurodevelopmental disorders?
The relative influence of genes and environment on the liability to neurodevelopmental disorders (NDDs) can be investigated using a twin design. This review highlights the results of the most recent twin studies of NDDs. Recent twin studies have confirmed that NDDs show moderate-to-high heritability, and that from an etiological viewpoint both autism spectrum disorder (ASD) and attention deficit hyperactivity disorder (ADHD) are best regarded as the extremes on a continuous liability distribution. Both ASD and ADHD show high heritability in childhood and a substantial drop in heritability in adulthood, which is likely explained by the use of different assessment strategies in childhood versus adulthood, or by a complex mechanism of gene-by-environment interaction. NDDs show substantial comorbidity among each other, and with other mental health problems, which is partly because of a shared genetic etiology between different disorders. The findings of twin studies implicate substantial heritability of NDDs, and warrant large-scale molecular genetic studies for such traits.


Achieving conservation science that bridges the knowledge-action boundary.
There are many barriers to using science to inform conservation policy and practice. Conservation scientists wishing to produce management-relevant science must balance this goal with the imperative of demonstrating novelty and rigor in their science. Decision makers seeking to make evidence-based decisions must balance a desire for knowledge with the need to act despite uncertainty. Generating science that will effectively inform management decisions requires that the production of information (the components of knowledge) be salient (relevant and timely), credible (authoritative, believable, and trusted), and legitimate (developed via a process that considers the values and perspectives of all relevant actors) in the eyes of both researchers and decision makers. We perceive 3 key challenges for those hoping to generate conservation science that achieves all 3 of these information characteristics. First, scientific and management audiences can have contrasting perceptions about the salience of research. Second, the pursuit of scientific credibility can come at the cost of salience and legitimacy in the eyes of decision makers, and, third, different actors can have conflicting views about what constitutes legitimate information. We highlight 4 institutional frameworks that can facilitate science that will inform management: boundary organizations (environmental organizations that span the boundary between science and management), research scientists embedded in resource management agencies, formal links between decision makers and scientists at research-focused institutions, and training programs for conservation professionals. Although these are not the only approaches to generating boundary-spanning science, nor are they mutually exclusive, they provide mechanisms for promoting communication, translation, and mediation across the knowledge-action boundary. We believe that despite the challenges, conservation science should strive to be a boundary science, which both advances scientific understanding and contributes to decision making.


Magnetic resonance imaging of navicular bursa adhesions.
Adhesions occur in the navicular bursa between the deep digital flexor tendon (DDFT) and other structures. Our objectives were to describe the appearance of navicular bursa adhesions on high-field magnetic resonance (MR) images, to compare these findings to findings at navicular bursoscopy, and to determine the prevalence of lesions in the remainder of the podotrochlear apparatus. Sixteen forelimbs from 14 horses that underwent MR imaging and navicular bursoscopy were evaluated. Adhesions were considered type 1 when characterized by a discontinuity in the navicular bursa fluid signal between two structures, type 2 when the navicular bursa fluid signal was disrupted and ill-defined tissue was present between two structures, and type 3 when the fluid signal was disrupted and well-defined tissue was present between two structures. Twenty-six adhesions were suspected on MR images and nineteen were visualized at surgery. The positive predictive value was 50% for type 1 adhesions, 67% for type 2 adhesions, and 100% for type 3 adhesions. Additional lesions were detected in the navicular bursa in 15 limbs, the DDFT in 13, the navicular bone in 15, the collateral sesamoidean ligaments in 9, and the distal sesamoidean impar ligament in 8. A discontinuity in the navicular bursa fluid signal with well-defined tissue between two structures detected on high-field MR images is diagnostic for a navicular bursa adhesion. Additional lesions in the podotrochlear apparatus are common in horses with navicular bursa adhesions.


A disease severity scale for systemic sclerosis: development and testing.
To develop and test a severity scale for individual organ involvements in systemic sclerosis (SSc, scleroderma). An international study group completed the following tasks: (1) developed a glossary of terms including all pertinent variables for 9 potentially affected organ systems; (2) collected prospective data to determine the feasibility and practicality of each proposed variable; (3) revised the initial list of variables; (4) determined the association of each variable with mortality (a proxy for morbidity) using 579 patients in an existing comprehensive longitudinal scleroderma databank; (5) developed a severity grading scale for each organ system by discussion and consensus; and (6) externally validated the scale using an independent group of 680 patients from the same databank. Nine organ-specific severity scales were developed from 0 (no documented involvement) to 4 (endstage disease). The data required for scale completion are relatively easy and practical for all physicians to obtain. This preliminary severity scale will be useful for assessing disease severity status in individual patients both at one point in time and longitudinally. The severity scale will assist in the design and conduct of clinical trials and the comparison of study populations with one another. The scale will serve as a framework for developing a scleroderma disease activity index.


Frontoethmoidal meningoencephalocoele repair in Cambodia: outcomes and cost comparisons.
In Cambodia, spina bifida is rare, but frontoethmoidal meningoencephalocoeles (MECs) are common. Mean life expectancy for patients with congenital MECs may be <20 years, but the complex treatment required has not been available in the country until recently. During visits by combined neurosurgical/craniofacial teams from both Germany and France, a method of repair has been developed that is suitable for the local conditions, affordable and has allowed Cambodian surgeons to learn how to successfully treat MECs. The surgical technique and initial results with 30 patients have been described in a previous publication. This paper presents the outcomes of 128 cases and illustrates that it is cost-effective for these patients to be treated in Cambodia.


A comparison of two computer access systems for functional text entry.
Functional written communication, an important goal in the rehabilitation of persons with tetraplegia, frequently is met through the use of personal computers and alternative computer access systems. To make informed decisions about alternative access systems, the therapist needs information on the efficacy of the available choices. The purpose of this study was to investigate the effectiveness of two commercially available systems for text entry, the traditional mouthstick and the Prentke Romich HeadMaster. Participants were a 25-year-old man and 76-year-old woman who both functioned at a C5 neurological level. Neither participant had previous experience with either system for text entry. A single-subject research design was used whereby Participant 1 experienced six phases of treatment (i.e., CBCBCB, where C = mouthstick and B = HeadMaster), and Participant 2 experienced four phases of treatment (i.e., BCBC). Participant 1 achieved a maximum rate of text entry of 5.85 wpm with both the HeadMaster and the mouthstick, whereas Participant 2 achieved a maximum rate of 7.15 wpm with the mouthstick and 4.85 wpm with the HeadMaster. Results from this study were similar to the results from previous comparison studies of persons with severe disabilities who had no experience with alternative computer access systems. Both participants were able to use both systems successfully; however, their respective rates of text entry were too slow to be functional in most employment situations.


A Technique to Identify Isoattenuating Gallstones with Dual-Layer Spectral CT: An ex Vivo Phantom Study.
Background Previously reported dual-energy CT methods for detecting noncalcified gallstones have reduced accuracy for gallstones smaller than 9 mm. Purpose To develop a dual-energy CT method for differentiating isoattenuating gallstones from bile and compare it with previously reported dual-energy CT methods by using a prospective ex vivo phantom reader study. Materials and Methods From May 2017 to May 2018, gallstones were collected from 105 patients (34 men; mean age, 51 years; age range, 18-84 years) undergoing cholecystectomy and placed inside 120-mL vials containing ox bile. The vials were placed inside a water-filled phantom and were scanned with dual-layer dual-energy CT. Thirty isoattenuating gallstones (4.3-24.7 mm in diameter) were evaluated. Conventional CT images, virtual noncontrast images, and monoenergetic images at 200 and 40 keV were created. Segmented images were created by using a two-dimensional histogram of Compton and photoelectric attenuation. Six readers evaluated the presence of isoattenuating gallstones in each image. Intra- and interreader agreement was measured by using percentage agreement, diagnostic performance was evaluated by using mean area under the receiver operating characteristic curve (AUC) estimates and pairwise comparisons, and the agreement of gallstone sizes measured at pathologic examination with those measured on segmented images was compared by using Bland-Altman analysis. Results For all gallstones, segmented images provided the highest mean intrareader (88.1%) and interreader (88.2% and 93.6%) agreements for all readers and reading sessions and the highest overall AUC (0.99; 95% confidence interval [CI]: 0.97, 1.00; adjusted P < .02 for all). For gallstones larger than 9 mm, no significant difference was found between the segmented and monoenergetic AUCs (all P > .94, adjusted P > .05 for all). For gallstones measuring 9 mm or smaller, the segmented images had the highest overall AUC (0.99; 95% CI: 0.97, 1.00; adjusted P < .01 for all). The mean difference in stone sizes was -0.6 mm, with limits of agreement from 2.6 to -3.8 mm. Conclusion Segmented images from Compton and photoelectric attenuation coefficients improve detection of isoattenuating gallstones compared with previously reported dual-energy CT methods. © RSNA, 2019 Online supplemental material is available for this article. See also the editorial by Matos in this issue.


Cellular mechanisms by which proinsulin C-peptide prevents insulin-induced neointima formation in human saphenous vein.
Endothelial cells (ECs) and smooth muscle cells (SMCs) play key roles in the development of intimal hyperplasia in saphenous vein (SV) bypass grafts. In diabetic patients, insulin administration controls hyperglycaemia but cardiovascular complications remain. Insulin is synthesised as a pro-peptide, from which C-peptide is cleaved and released into the circulation with insulin; exogenous insulin lacks C-peptide. Here we investigate modulation of human SV neointima formation and SV-EC and SV-SMC function by insulin and C-peptide. Effects of insulin and C-peptide on neointima formation (organ cultures), EC and SMC proliferation (cell counting), EC migration (scratch wound), SMC migration (Boyden chamber) and signalling (immunoblotting) were examined. A real-time RT-PCR array identified insulin-responsive genes, and results were confirmed by real-time RT-PCR. Targeted gene silencing (siRNA) was used to assess functional relevance. Insulin (100 nmol/l) augmented SV neointimal thickening (70% increase, 14 days), SMC proliferation (55% increase, 7 days) and migration (150% increase, 6 h); effects were abrogated by 10 nmol/l C-peptide. C-peptide did not affect insulin-induced Akt or extracellular signal-regulated kinase signalling (15 min), but array data and gene silencing implicated sterol regulatory element binding transcription factor 1 (SREBF1). Insulin (1-100 nmol/l) did not modify EC proliferation or migration, whereas 10 nmol/l C-peptide stimulated EC proliferation by 40% (5 days). Our data support a causative role for insulin in human SV neointima formation with a novel counter-regulatory effect of proinsulin C-peptide. Thus, C-peptide can limit the detrimental effects of insulin on SMC function. Co-supplementing insulin therapy with C-peptide could improve therapy in insulin-treated patients.


Avoidance of hydrolyzed casein by mice.
When casein, a milk protein, is hydrolyzed, it renders human foods that contain it (e.g., hypoallergenic infant formula, cheeses) distasteful to many people. This rejection of hydrolyzed casein (HC)-containing foods has recently been found to also occur in a non-human species (deer, Odocoileus spp.). Identifying other animals that avoid HC would facilitate understanding how and why HC-containing food is often rejected. This study determined whether HC-containing food is avoided by Mus musculus and whether consumption patterns were sensitive to testing conditions, specifically food form (powder, pellet or dough) and food access (ad libitum or 1.5 h/day following 6 h of food deprivation). Diets were offered in two-choice tests that paired an HC-containing food with an intact casein-containing alternative at seven protein concentrations (0%-50% w/w). Five experimental groups were tested under different combinations of food form and food access. Three groups (ad lib/powder, ad lib/pellet, and 1.5 h/pellet) avoided the HC diet starting at the 30% protein level. At the 40% and 50% protein levels, all groups showed strong avoidance of HC. Although testing conditions influenced total caloric intake and body weight gain, avoidance of HC at the highest concentrations was robust to the manipulations in experimental conditions. Our study suggests that mice may be a useful model for understanding the mechanisms of HC rejection.


Biodegradation of gasoline by gellan gum-encapsulated bacterial cells.
Encapsulated cell bioaugmentation is a novel alternative solution to in situ bioremediation of contaminated aquifers. This study was conducted to evaluate the feasibility of such a remediation strategy based on the performance of encapsulated cells in the biodegradation of gasoline, a major groundwater contaminant. An enriched bacterial consortium, isolated from a gasoline-polluted site, was encapsulated in gellan gum microbeads (16-53 microm diameter). The capacity of the encapsulated cells to degrade gasoline under aerobic conditions was evaluated in comparison with free (non-encapsulated) cells. Encapsulated cells (2.6 mg(cells) x g(-1) bead) degraded over 90% gasoline hydrocarbons (initial concentration 50-600 mg x L(-1)) within 5-10 days at 10 degrees C. Equivalent levels of free cells removed comparable amounts of gasoline (initial concentration 50-400 mg x L(-1)) within the same period but required up to 30 days to degrade the highest level of gasoline tested (600 mg x L(-1)). Free cells exhibited a lag phase in biodegradation, which increased from 1 to 5 days with an increase in gasoline concentration (200-600 x mg L(-1)). Encapsulation provided cells with a protective barrier against toxic hydrocarbons, eliminating the adaptation period required by free cells. The reduction of encapsulated cell mass loading from 2.6 to 1.0 mg(cells) x g(-1) bead caused a substantial decrease in the extent of biodegradation within a 30-day incubation period. Encapsulated cells dispersed within the porous soil matrix of saturated soil microcosms demonstrated a reduced performance in the removal of gasoline (initial concentrations of 400 and 600 mg x L(-1)), removing 30-50% gasoline hydrocarbons compared to 40-60% by free cells within 21 days of incubation. The results of this study suggest that gellan gum-encapsulated bacterial cells have the potential to be used for biodegradation of gasoline hydrocarbons in aqueous systems.


Active and passive frequency stabilization for a Q-switched Z-fold radio-frequency-excited waveguide CO2 laser with two channels.
We report on passive offset frequency stability in free-running applications and active offset frequency stability achieved by use of a frequency-locking technique for a Q-switched Z-fold rf-excited waveguide CO2 laser with two channels. The laser structure of common electrodes and two channels has the advantage of compensating for the frequency variation caused by variations in temperature, cavity length, gas refractive index, and mechanical vibrations, so its offset frequency stability is higher than that of two separate lasers. In the experiments, the offset frequency shift was less than 6 MHz for 3 min in free-running mode. The technique of active offset frequency locking by counting was also introduced. The beat frequency shifting value was smaller than +/- 0.5 MHz in the long-term.


Novel insights into the echinoderm nervous system from histaminergic and FMRFaminergic-like cells in the sea cucumber Leptosynapta clarki.
Understanding of the echinoderm nervous system is limited due to its distinct organization in comparison to other animal phyla and by the difficulty in accessing it. The transparent and accessible, apodid sea cucumber Leptosynapta clarki provides novel opportunities for detailed characterization of echinoderm neural systems. The present study used immunohistochemistry against FMRFamide and histamine to describe the neural organization in juvenile and adult sea cucumbers. Histaminergic- and FMRFaminergic-like immunoreactivity is reported in several distinct cell types throughout the body of L. clarki. FMRFamide-like immunoreactive cell bodies were found in the buccal tentacles, esophageal region and in proximity to the radial nerve cords. Sensory-like cells in the tentacles send processes toward the circumoral nerve ring, while unipolar and bipolar cells close to the radial nerve cords display extensive processes in close association with muscle and other cells of the body wall. Histamine-like immunoreactivity was identified in neuronal somatas located in the buccal tentacles, circumoral nerve ring and in papillae distributed across the body. The tentacular cells send processes into the nerve ring, while the processes of cells in the body wall papillae extend to the surface epithelium and radial nerve cords. Pharmacological application of histamine produced a strong coordinated, peristaltic response of the body wall suggesting the role of histamine in the feeding behavior. Our immunohistochemical data provide evidence for extensive connections between the hyponeural and ectoneural nervous system in the sea cucumber, challenging previously held views on a clear functional separation of the sub-components of the nervous system. Furthermore, our data indicate a potential function of histamine in coordinated, peristaltic movements; consistent with feeding patterns in this species. This study on L. clarki illustrates how using a broader range of neurotransmitter systems can provide better insight into the anatomy, function and evolution of echinoderm nervous sytems.


Mobile phone text messaging intervention to improve alertness and reduce sleepiness and fatigue during shiftwork among emergency medicine clinicians: study protocol for the SleepTrackTXT pilot randomized controlled trial.
Mental and physical fatigue while at work is common among emergency medical services (EMS) shift workers. Extended shifts (for example 24 hours) and excessive amounts of overtime work increase the likelihood of negative safety outcomes and pose a challenge for EMS fatigue-risk management. Text message-based interventions are a potentially high-impact, low-cost platform for sleep and fatigue assessment and distributing information to workers at risk of negative safety outcomes related to sleep behaviors and fatigue. We will conduct a pilot randomized trial with a convenience sample of adult EMS workers recruited from across the United States using a single study website. Participants will be allocated to one of two possible arms for a 90-day study period. The intervention arm will involve text message assessments of sleepiness, fatigue, and difficulty with concentration at the beginning, during, and end of scheduled shifts. Intervention subjects reporting high levels of sleepiness or fatigue will receive one of four randomly selected intervention messages promoting behavior change during shiftwork. Control subjects will receive assessment only text messages. We aim to determine the performance characteristics of a text messaging tool for the delivery of a sleep and fatigue intervention. We seek to determine if a text messaging program with tailored intervention messages is effective at reducing perceived sleepiness and/or fatigue among emergency medicine clinician shift workers. Additional aims include testing whether a theory-based behavioral intervention, delivered by text message, changes 'alertness behaviors'. The SleepTrackTXT pilot trial could provide evidence of compliance and effectiveness that would support rapid widespread expansion in one of two forms: 1) a stand-alone program in the form of a tailored/individualized sleep monitoring and fatigue reduction support service for EMS workers; or 2) an add-on to a multi-component fatigue risk management program led and maintained by employers or by safety and risk management services. Clinicaltrials.gov NCT02063737, Registered on 10 January 2014.


Consumer acceptance of dairy products with a saturated fatty acid-reduced, monounsaturated fatty acid-enriched content.
Agriculture-based reformulation initiatives, including oleic acid-rich lipid supplementation of the dairy cow diet, provide a novel means for reducing intake of saturated fatty acids (SFA) at a population level. In a blinded manner, this study evaluated the consumer acceptance of SFA-reduced, monounsaturated fatty acid-enriched (modified) milk, Cheddar cheese, and butter when compared with control and commercially available comparative samples. The effect of providing nutritional information about the modified cheese was also evaluated. Consumers (n = 115) rated samples for overall liking (appearance, flavor, and texture) using 9-point hedonic scales. Although no significant differences were found between the milk samples, the modified cheese was liked significantly less than a regular-fat commercial alternative for overall liking and liking of specific modalities and had a lower liking of texture score compared with the control cheese. The provision of health information significantly increased the overall liking of the modified cheese compared with tasting the same sample in a blinded manner. Significant differences were evident between the butter samples for overall liking and modalities of liking; all of the samples were significantly more liked than the commercial butter and sunflower oil spread. In conclusion, this study illustrated that consumer acceptance of SFA-reduced, monounsaturated fatty acid-enriched dairy products was dependent on product type. Future research should consider how optimization of the textural properties of fatty acid-modified (and fat-reduced) cheese might enhance consumer acceptance of this product.


Exploration of risk factors for high-risk adverse events in elderly patients after discharge and comparison of discharge planning screening tools.
Discharge planning is an effective strategy to prevent adverse health events and reduce medical expenditures. The high-risk target populations of discharged elderly patients and important predictors for the occurrence of adverse events are still not clear. Therefore, the purposes of this study were to examine the validity of discharge planning screening tools in sufficiently identifying high-risk adverse events to health after discharge and to compare two screening tools with our study model. We conducted a prospective study and recruited elderly patients who had had no hospitalization within 3 months before admission to 13 general wards of a medical center in northern Taiwan from November 2018 to May 2020. Elderly patients were randomly selected during the study period. Within 24 h of admission, patients were asked to consent to join this study. After the patient was discharged, the patient's health and hospitalization for the next year were tracked by telephone interviews. In total, 300 participants were recruited for this study. Incidences of high-risk adverse events within 30 days, 60 days, and 12 months after discharge were 20.3%, 25.7%, and 48.7% respectively. A logistic regression showed that an increased age, physical or mental disabilities or a major illness, a low body-mass index, and having been hospitalized in the past year were significantly related to the occurrence of high-risk events among elderly discharge patients. The pooled sensitivity of the Pra was 52% and the specificity was 72%; the pooled sensitivity of the LACE index was 67% and the specificity was 36%. The predictive model of this study had a higher discriminatory power than the Pra and LACE index for high-risk events after discharge. Elderly patients are more vulnerable to high-risk adverse events after discharge. Both the LACE index and Pra are useful discharge planning screening tools to screen for high-risk adverse events after discharge. Elderly patients need more-active and complete continuity of care plans and discharge planning services to ensure that the overall quality of patient care can be improved and readmissions and mortality reduced. The findings of this study can provide information for discharge planning managers to identify high-risk elderly patients during hospitalization and promptly offer care education or resources to improve care management.


Novel gene targets for miRNA146a and miRNA155 in anterior uveitis.
Anterior uveitis (AU) is the most common form of intraocular inflammation. MicroRNAs (miRNA) are small, non-coding RNAs functioning as post-transcriptional repressors of gene expression. Knowledge of miRNAs can implicate specific genes and pathogenic signalling pathways in disease. This study examines miRNA expression, function and target genes in AU pathogenesis. AU and healthy control (HC) peripheral blood mononuclear cells (PBMC) were initially screened for expression of five miRNAs by real-time PCR. Regulation of the aberrantly expressed miRNAs by TLR1/2, TLR3, TLR4, IL1β and TNFα was quantified by real-time PCR and paired cytokine outputs measured by ELISA. Functional effects of miRNA overexpression using transfected THP1 cells examined IL6, IL8, IL10 and IL1β cytokine outputs by ELISA. Target genes were identified using TargetScan online computational algorithm and relevant targets verified by cloning of the 3'UTR and luciferase reporter gene assays. Increased expression of miRNA146a (p<0.01), miRNA155 (p<0.05) and miRNA125a5p (p<0.01) was demonstrated in AU PBMC compared with HC. miRNA155 was increased following TLR1/2 (p<0.05) and TLR4 (p<0.05) stimulation and miRNA146a increased in response to IL1β (p<0.05). In a proinflammatory environment, miRNA155 overexpression in THP1 cells yielded increased cytokine output whereas miRNA146a overexpression showed decreased cytokine output. CD80, PRKCE and VASN were confirmed as novel targets for miRNA146a and SMAD2, TYRP1 and FBXO22 for miRNA155. This study identifies overexpression of proinflammatory miRNA155, regulatory miRNA146a and miRNA125a-5p in AU. CD80, PRKCE and VASN are novel miRNA146a targets and SMAD2, TYRP1 and FBXO22 are novel targets for miRNA155.