Diagnostic Yield of Endoscopic Ultrasound in Common Bile Duct Dilation: A Real Breakthrough.

BACKGROUND: Endoscopic ultrasound (EUS) is important for the evaluation of patients with common bile duct (CBD) dilation. AIMS: The purpose of this study was to evaluate the diagnostic performance of EUS for CBD dilation in patients with negative initial studies. METHODS: This was a retrospective cohort study that included patients who underwent EUS for CBD dilation (≥ 7 mm if intact anatomy or ≥ 10 mm if prior cholecystectomy) in the absence of pathology on previous ultrasonography (US), computed tomography (CT), and/or magnetic resonance cholangiopancreatography (MRCP). RESULTS: A total of 109 patients were included, among whom 41 had a positive EUS: 33 choledocholithiasis (30.3%), 6 chronic pancreatitis (5.5%), and 2 ampullary cancer (1.8%). If the EUS was negative, no pathology was found during 1-year follow-up. Older age was associated with positive EUS (79 versus 71 years, p = 0.030). Patients with jaundice, cholelithiasis, and altered liver biochemistry were 16.2 (p = 0.002), 3.1 (p = 0.024), and 2.9 (p = 0.009) times more likely to have positive EUS, respectively. A total of 53 patients had a negative MRCP (48.6%); those with biliary abdominal pain and jaundice were 15.5 (p < 0.001) and 20.0 (p = 0.007) times more likely to have positive EUS, respectively. Considering asymptomatic patients with normal liver tests, CBD diameter ≥ 10 mm in US and ≥ 11 mm in CT can predict a positive EUS (AUC 0.754, p = 0.047 and AUC 0.734, p = 0.048). CONCLUSIONS: EUS is a useful diagnostic method for patients with unexplained CBD dilation, even if negative MRCP, and especially in patients with older age, abdominal pain, jaundice, cholelithiasis, and/or altered liver biochemistry. CBD diameter in US and CT had a moderate discriminative ability in predicting positive EUS in asymptomatic patients without altered liver biochemistry.

Validity Evidence for the Brief Self-Control Scale in the Spanish Adult Population: A Systematic Study.

The Brief Self-Control Scale (BSCS) has been widely studied across languages and populations. However, research examining the Spanish version is scant and limited to the adolescent population. Our aim here was to provide validity evidence for use of the BSCS with Spanish adults by analyzing and comparing the psychometric properties of different versions of the scale (13-item, 10-item, 9-item, 8-item, and 7-item). Confirmatory factor analysis was used to test the internal structure based on one-factor and two-factor models. Results obtained in a sample of 676 Spanish adults showed adequate fit indices for the two-factor structure of the 9-item, 8-item, and 7-item versions of the BSCS, although only the 9-item and 8-item BSCS were found to be invariant across gender. Item homogeneity and reliability of factor scores for these two versions (9-item and 8-item) were satisfactory. We also provide novel validity evidence based on relationships with indicators of psychological adjustment and wellbeing. Scores on the 9-item and 8-item BSCS correlated with life satisfaction, flourishing, self-esteem, distress, depression, and loneliness, and hence both may be suitable for use in mental health assessment contexts.

Secondary bruxism: A valid diagnosis or just a coincidental finding of additional masticatory muscle activity? A narrative review of literature.

INTRODUCTION: Bruxism is defined as a repetitive masticatory muscle activity that can manifest it upon awakening (awake bruxism-AB) or during sleep (sleep bruxism-SB). Some forms of both, AB and SB can be associated to many other coexistent factors, considered of risk for the initiation and maintenance of the bruxism. Although controversial, the term 'secondary bruxism' has frequently been used to label these cases. The absence of an adequate definition of bruxism, the non-distinction between the circadian manifestations and the report of many different measurement techniques, however, are important factors to be considered when judging the literature findings. The use (and abuse) of drugs, caffeine, nicotine, alcohol and psychoactive substances, the presence of respiratory disorders during sleep, gastroesophageal reflux disorders and movement, neurological and psychiatric disorders are among these factors. The scarcity of controlled studies and the complexity and interactions among all aforementioned factors, unfortunately, does not allow to establish any causality or temporal association with SB and AB. The supposition that variables are related depends on different parameters, not clearly demonstrated in the available studies. OBJECTIVES: This narrative review aims at providing oral health care professionals with an update on the co-risk factors and disorders possibly associated with bruxism. In addition, the authors discuss the appropriateness of the term 'secondary bruxism' as a valid diagnostic category based on the available evidence. CONCLUSION: The absence of an adequate definition of bruxism, the non-distinction between the circadian manifestations and the report of many different measurement techniques found in many studies preclude any solid and convincing conclusion on the existence of the 'secondary' bruxism.

Ultrasonographic scores for ileal Crohn's disease assessment: Better, worse or the same as contrast-enhanced ultrasound?

BACKGROUND: Intestinal ultrasound (IUS) is an increasingly used non-invasive tool to evaluate Crohn's disease (CD) activity. Recently, two IUS scores that evaluate inflammatory activity have emerged: the Simple Ultrasound Activity Score for CD (SUS-CD) and the International Bowel Ultrasound Segmental Activity Score (IBUS-SAS). We aimed to compare the accuracy of SUS-CD, IBUS-SAS and contrast-enhanced ultrasound (CEUS) in predicting inflammatory activity in the terminal ileum in ileocolonoscopy in CD patients. METHODS: Retrospective study including all consecutive CD patients submitted to IUS with CEUS directed to the terminal ileum performed by a single operator between April 2016 and March 2020. Segmental SUS-CD and IBUS-SAS were calculated. A time-intensity curve of the contrast bowel wall enhancement was created with measurement of peak intensity using CEUS. The CD endoscopic activity in ileocolonoscopy was graded by Simple Endoscopic Score for CD (SES-CD) as inactive (SES-CD < 7) or active (SES-CD ≥ 7). RESULTS: Fifty patients were included, 54.0% were female, with mean age of 34 ± 12 years, and most had isolated ileal disease (60.0%), and a nonstricturing, nonpenetrating behaviour (44.0%). Most of the patients (60.0%) had active endoscopic disease (SES-CD ≥ 7). SUS-CD and IBUS-SAS were not different between patients with active or inactive endoscopic disease (p = 0.15; 0.57, respectively), having a poor accuracy to correlate endoscopic activity (area under de curve (AUC) 0.62; 0.55, respectively). Peak intensity in CEUS was significantly different in patients with active or inactive endoscopic disease (p = 0.004), having a good accuracy to correlate endoscopic activity (AUC 0.80). CONCLUSION: Unlike CEUS, SUS-CD and IBUS-SAS were not able to accurately correlate endoscopic activity in terminal ileum in CD. Therefore, CEUS is a non-invasive emerging method that should be increasingly integrated in the ultrasonographic evaluation of CD patients.

Cognition in informal caregivers: evidence from an English population study.

BACKGROUND AND OBJECTIVES: The relationship between caregiving and cognition remains unclear. We investigate this association comparing four cognitive tasks and exploring the role of potential explanatory pathways such as healthy behaviours (healthy caregiver hypothesis) and depression (stress process model). RESEARCH DESIGN AND METHODS: Respondents were from English Longitudinal Study of Ageing (ELSA) (N = 8910). Cognitive tasks included immediate and delayed word recall, verbal fluency and serial 7 subtraction. Series of hierarchical linear regressions were performed. Adjustments included socio-demographics, health related variables, health behaviours and depression. RESULTS: Being a caregiver was positively associated with immediate and delayed recall, verbal fluency but not with serial 7. For immediate and delayed recall, these associations were partially attenuated when adjusting for health behaviours, and depression. For verbal fluency, associations were partially attenuated when adjusting for depression but fully attenuated when adjusting for health behaviours. No associations were found for serial 7. DISCUSSION AND IMPLICATIONS: Our findings show that caregivers have higher level of memory and executive function compared to non-caregivers. For memory, we found that although health behaviours and depression can have a role in this association, they do not fully explain it. However, health behaviours seem to have a clear role in the association with executive function. Public health and policy do not need to target specifically cognitive function but other areas as the promotion of healthy behaviours and psychological adjustment such as preventing depression and promoting physical activity in caregivers.

Comparison of Stud- Retentor Versus Bar-Clip Attachment as Implant- Supported Systems Used in Overdentures: A Systematic Review and Meta-Analysis.

This study aimed to perform a systematic review and meta-analysis to analyze the results obtained clinically for bar-clip versus stud-retainers in overdentures. Three databases (PubMed Central, MEDLINE, and BvSalud) were used beyond a manual search. The study followed strictly the inclusion and exclusion criteria, considering the PICO strategy. For the risk of bias and quality assessment of studies, in the case of RCT, there were six domains of analysis, and for non-RCT studies, the Modified Newcastle-Ottawa Scale was performed. A meta-analysis was developed using the available data for marginal bone loss (MBL) and survival rate. 25 studies were included. The stud-retentor had the lowest implant SR (87.6%) and the greatest MBL (1.96 mm). For the bar-clip system, the mean survival rate was 95.91%, with only 4 studies included for this system, and the mean MBL was 1.13 mm. Only 3 studies directly compared both systems quantitatively, showing a significantly greater MBL toward the stud-retention group. The results may not allow determination of the best system for overdenture (stud retentor or bar-clip). Therefore, most of the studies suggested the stud-retentor as a more preferable system due to better distribution of forces, biological peri-implant behavior, low-cost, and ease for removal facilitating the sanitization and/or repair.

The potential of using climate indices as powerful tools to explain mortality anomalies: An application to mainland Spain.

Changes in the frequency and magnitude of extreme weather events represent one of the key indicators of climate change and variability. These events can have an important impact on mortality rates, especially in the ageing population. This study assessed the spatial and seasonal distributions of mortality rates in mainland Spain and their association with climatic conditions over the period 1979-2016. The analysis was done on a seasonal and annual basis using 79 climatic indices and regional natural deaths data. Results indicate large spatial variability of natural deaths, which is mostly related to how the share of the elderly in the population varied across the studied regions. Spatially, both the highest mortality rates and the largest percentage of elders were found in the northwest areas of the study domain, where an extreme climate prevails, with very cold winters and hot summers. A strong seasonality effect was observed, winter shows more than 10% of natural deaths compared to the rest of the seasons. Also, results suggest a strong relation between climatic indices and natural deaths, albeit with a high spatial and seasonal variability. Climatic indices and natural deaths show a stronger correlation in winter and summer than in spring and autumn.

The role of character strengths in predicting gains in informal caregivers of dementia.

OBJECTIVES: Although providing care to a person with dementia can have a negative impact, caregivers may also perceive certain benefits and gains through the tasks they perform. Our aim here was to study caregiver gains within the framework of positive psychology, exploring the predictive power of character strengths, while controlling for sociodemographic variables and variables related to the dementia and caring. METHODS: A sample of 105 main caregivers of people diagnosed with dementia completed a sociodemographic questionnaire, the Gain in Alzheimer care Instrument and the Values in Action Inventory of Strengths. Correlational analysis and hierarchical regression were conducted. RESULTS: Eighteen character strengths were positively and significantly correlated with gain scores. Regression analysis indicated that level of education was negatively related to gain scores. In addition, hope was the character strength which best predicted the gain score, such that caregivers who scored higher on hope tended to perceive greater benefits from their role. CONCLUSION: The results suggest that hope may play an important role in relation to the perceived gains of caregiving. Intervention programmes based on positive psychology and aimed at enhancing character strengths, especially hope, could help caregivers to identify the positive aspects of their caring role.

Mathematical model describing CoViD-19 in São Paulo, Brazil - evaluating isolation as control mechanism and forecasting epidemiological scenarios of release.

In São Paulo, Brazil, the first case of coronavirus disease 2019 (CoViD-19) was confirmed on 26 February, the first death due to CoViD-19 was registered on 16 March, and on 24 March, São Paulo implemented the isolation of persons in non-essential activities. A mathematical model was formulated based on non-linear ordinary differential equations considering young (60 years old or less) and elder (60 years old or more) subpopulations, aiming to describe the introduction and dissemination of the new coronavirus in São Paulo. This deterministic model used the data collected from São Paulo to estimate the model parameters, obtaining R0 = 6.8 for the basic reproduction number. The model also allowed to estimate that 50% of the population of São Paulo was in isolation, which permitted to describe the current epidemiological status. The goal of isolation implemented in São Paulo to control the rapid increase of the new coronavirus epidemic was partially succeeded, concluding that if isolation of at least 80% of the population had been implemented, the collapse in the health care system could be avoided. Nevertheless, the isolated persons must be released one day. Based on this model, we studied the potential epidemiological scenarios of release by varying the proportions of the release of young and elder persons. We also evaluated three different strategies of release: All isolated persons are released simultaneously, two and three releases divided in equal proportions. The better scenarios occurred when young persons are released, but maintaining elder persons isolated for a while. When compared with the epidemic without isolation, all strategies of release did not attain the goal of reducing substantially the number of hospitalisations due to severe CoViD-19. Hence, we concluded that the best decision must be postponing the beginning of the release.

Identifying pulmonary nodules or masses on chest radiography using deep learning: external validation and strategies to improve clinical practice.

AIM: To test the diagnostic performance of a deep learning-based system for the detection of clinically significant pulmonary nodules/masses on chest radiographs. MATERIALS AND METHODS: Using a retrospective study of 100 patients (47 with clinically significant pulmonary nodules/masses and 53 control subjects without pulmonary nodules), two radiologists verified clinically significantly pulmonary nodules/masses according to chest computed tomography (CT) findings. A computer-aided diagnosis (CAD) software using a deep-learning approach was used to detect pulmonary nodules/masses to determine the diagnostic performance in four algorithms (heat map, abnormal probability, nodule probability, and mass probability). RESULTS: A total of 100 cases were included in the analysis. Among the four algorithms, mass algorithm could achieve a 76.6% sensitivity (36/47, 11 false negative) and 88.68% specificity (47/53, six false-positive) in the detection of pulmonary nodules/masses at the optimal probability score cut-off of 0.2884. Compared to the other three algorithms, mass probability algorithm had best predictive ability for pulmonary nodule/mass detection at the optimal probability score cut-off of 0.2884 (AUCMass: 0.916 versus AUCHeat map: 0.682, p<0.001; AUCMass: 0.916 versus AUCAbnormal: 0.810, p=0.002; AUCMass: 0.916 versus AUCNodule: 0.813, p=0.014). CONCLUSION: In conclusion, the deep-learning based computer-aided diagnosis system will likely play a vital role in the early detection and diagnosis of pulmonary nodules/masses on chest radiographs. In future applications, these algorithms could support triage workflow via double reading to improve sensitivity and specificity during the diagnostic process.

Differences in multi-echo chemical shift encoded MRI proton density fat fraction estimation based on multifrequency fat peaks selection in non-alcoholic fatty liver disease patients.

AIM: To compare the performance of multi-echo chemical-shift-encoded (MECSE) magnetic resonance imaging (MRI) proton density fat fraction (PDFF) estimation, considering three different fat frequency peak combinations, for the quantification of steatosis in patients with non-alcoholic fatty liver disease (NAFLD). MATERIALS AND METHODS: The present study was a prospective cross-sectional research of 121 patients with metabolic syndrome and evidence of hepatic steatosis on ultrasound, who underwent a 3 T MRI examination. All patients were studied with a multifrequency MECSE sequence. The PDFF was calculated using six peaks (MECSEp123456), three peaks (MECSEp456), and a single peak (MECSEp5) model. The two simpler fat peak models were compared to the six peaks model, which was considered the reference standard. Linearity was evaluated using linear regression while agreement was described using Bland-Altman analysis. RESULTS: The mean age was 47 (±9) years and BMI was 29.9 (±2.9) kg/m2. Steatosis distribution was 15%/31%/54% (S1/S2/S3, respectively). Compared to MECSEp123456, both models provided linear PDFF measurements (R2= 0.99 and 0.97, MECSEp456 and MECSEp5 respectively). Regression slope (0.92; p<0.001) and mean Bland-Altman bias (-1.5%; 95% limits of agreement: -3.19%, 0.22%) indicated minimal underestimation by using PDFF-MECSEp456. Nonetheless, mean differences in PDFF estimations varied from -1.5% (MECSEp456,p=0.006) to -2.2% (MECSEp5,p<0.001) when compared to full six fat frequencies model. CONCLUSION: Although simpler spectral fat MECSE analysis shows a linear relationship with the standard six peaks model, their variation in estimated PDFF values introduces a low but clinically significant bias in fat quantification and steatosis grading in NAFLD patients.

Aerobic processes for bioleaching manganese and silver using microorganisms indigenous to mine tailings.

Mining wastes containing appreciable concentrations of silver are considered alternative sources for metal extraction, although these wastes are often refractory due to the presence of manganese oxides. The high cost and/or environmental impact of the hydrometallurgical and pyrometallurgical extraction processes make it necessary to search for biotechnological processes for the solubilization of manganese compounds. This paper describes the characterization of chemoorganotrophic microorganisms indigenous to the tailings of a silver mine located in Coahuila, México, regarding their capability to remove manganese and silver present in these residues by lixiviation. The Bacterial and fungal strains isolated were identified by sequencing the rDNA 16S and ITS-1-ITS-2 genomic regions, respectively; the bacterial strains correspond to isolates of Roseospira sp. and Sphingomonas sp., whereas the fungal strains include isolates of Cladosporium sp. A, Cladosporium sp. B and Penicillium chrysogenum. These fungal strains show an effective capacity to lixiviate manganese and silver from solid mine residue when incubated in 9 k medium; it was found that under these conditions, leaching of metals occurs due to a mixed biotic-abiotic process, which yields manganese and silver leaching efficiencies in the ranges of 58-74% and 40-67%, respectively. The fungal strains grown in the LMM medium and the bacterial strains incubated in the PDB medium caused leaching of manganese with a lower efficiency in the range of 0.17-0.24% and 1.42-1.73%, respectively; under these conditions, silver leaching by fungal and bacterial strains appeared to be reduced (< 0.1%).Through in vitro cultures, it was determined that P. chrysogenum and Sphingomonas sp. showed the highest levels of silver resistance.

Yeast derivatives and wheat germ in the adult diet modulates fecundity in a tephritid pest.

Anastrepha fraterculus (Wiedemann), a pest of great economic importance in South America, needs urgently to be controlled by environmentally friendly methods such as the sterile insect technique for which mass rearing of insects is required. Because oogenesis takes place during the adult stage, mass-rearing facilities should provide the females a diet that maximizes egg production at the lowest cost. Accordingly, we investigated the effect of artificial protein sources in the adult diet (yeast derivatives of different cost but with similar amino acids profiles, and the addition of wheat germ) on fecundity. Additionally, we evaluated different ratios of yeast derivatives or wheat germ on ovary maturation, fecundity, and fertility as well as their association with the nutrient content of females. Females fed hydrolyzed yeast and yeast extract attained the highest fecundity level, and those fed brewer's yeast the lowest. Reducing the amount of hydrolyzed yeast, an expensive protein source, in the diet negatively affected fecundity and ovary maturation. Increasing the amount of brewer's yeast, a low-cost protein source, did not favor fecundity. The addition of wheat germ in the adult diet improved fecundity regardless of the yeast derivate considered. Percentage of egg hatch was not affected by the diet. Nutrient content of A. fraterculus females varied according to the adult diet provided and mating status. Our findings provide novel baseline information to understand the role of nutrition on reproductive performance of A. fraterculus females and are discussed in the context of resource allocation. They also provide valuable advances in the search for cost-effective adult diets at fruit fly mass rearing facilities.

The expression level of SOX2 at the blastocyst stage regulates the developmental capacity of bovine embryos up to day-13 of in vitro culture.

Quality of in vitro-produced embryos is influenced by changes in gene expression in response to adverse conditions. Gene markers for predicting 'good embryos' do not exist at present. We propose that the expression of pluripotency markers OCT4-SOX2-NANOG in D9 (day 9) bovine demi-embryos correlated with development at D13 (day 13). Day 8 in vitro-produced blastocysts were split in two cloned halves, one half (D9) was subjected to analysis of pluripotency markers and the other was kept in culture until D13 of development. Embryo development was scored and correlated with its own status at D9 and assigned to one of two categories: G1, arrested/dead; or G2, development up to D13. SOX2 and NANOG expression levels were significantly higher in embryos from G1 and there was also negative correlation between SOX2 and embryo survival to D13 (G3; r = -0.37; P = 0.03). We observed a significant reduction in the expression of the three studied genes from D9 to D13. Furthermore, there was a correlation between the expression of pluripotency markers at D9 and embryo diameter and the expression of trophoblastic markers at D13 (TP1-EOMES-FGF4-CDX2-TKDP1). Finally, the quotient between the relative expression of SOX2 and OCT4 in the D9 blastocysts from G1 and G2 showed that embryos that were considered as competent (G2) had a quotient close to one, while the other group had a quotient of 2.3 due to a higher expression of SOX2. These results might indicate that overexpression of SOX2 at the blastocyst stage had a negative effect on the control of embryonic developmental potential.

Neuroinflammation in obesity: circulating lipopolysaccharide-binding protein associates with brain structure and cognitive performance.

BACKGROUND/OBJECTIVES: Growing evidence implicates neuroinflammation in the pathogenesis of diet-induced obesity and cognitive dysfunction in rodent models. Obesity is associated with reduced white matter integrity and cognitive decline. Circulating lipopolysaccharide binding protein (LBP) concentration is known to be increased in patients with obesity. Here, we aimed to evaluate whether circulating LBP is associated longitudinally with white matter structure and cognitive performance according to obesity status. SUBJECTS/METHODS: This longitudinal study analyzed circulating LBP (ELISA), DTI-metrics (axial diffusivity (L1), fractional anisotropy (FA) and radial diffusivity (RD)) in specific regions of the white matter of 24 consecutive middle-aged obese subjects (13 women) and 20 healthy volunteers (10 women) at baseline and two years later. Digit Span Test (DST) was used as a measure of working memory/short-term verbal memory. RESULTS: Circulating LBP concentration was associated with FA and L1 values of several white matter regions both at baseline and follow-up. The associations remained significant after controlling for age, BMI, fat mass and plasma high sensitivity C-reactive protein. Importantly, the increase in LBP over time impacted negatively on FA and L1 values and on DST performance. CONCLUSIONS: Circulating LBP associates with brain white matter integrity and working memory/short-term verbal memory in both obese and non-obese subjects.

Defective expression of apoptosis-related molecules in multiple sclerosis patients is normalized early after autologous haematopoietic stem cell transplantation.

Defective apoptosis might be involved in the pathogenesis of multiple sclerosis (MS). We evaluated apoptosis-related molecules in MS patients before and after autologous haematopoietic stem cell transplantation (AHSCT) using BCNU, Etoposide, AraC and Melphalan (BEAM) or cyclophosphamide (CY)-based conditioning regimens. Patients were followed for clinical and immunological parameters for 2 years after AHSCT. At baseline, MS patients had decreased proapoptotic BAD, BAX and FASL and increased A1 gene expression when compared with healthy counterparts. In the BEAM group, BAK, BIK, BIMEL , FAS, FASL, A1, BCL2, BCLXL , CFLIPL and CIAP2 genes were up-regulated after AHSCT. With the exception of BIK, BIMEL and A1, all genes reached levels similar to controls at day + 720 post-transplantation. Furthermore, in these patients, we observed increased CD8+ Fas+ T cell frequencies after AHSCT when compared to baseline. In the CY group, we observed increased BAX, BCLW, CFLIPL and CIAP1 and decreased BIK and BID gene expressions after transplantation. At day + 720 post-AHSCT, the expression of BAX, FAS, FASL, BCL2, BCLXL and CIAP1 was similar to that of controls. Protein analyses showed increased Bcl-2 expression before transplantation. At 1 year post-AHSCT, expression of Bak, Bim, Bcl-2, Bcl-xL and cFlip-L was decreased when compared to baseline values. In summary, our findings suggest that normalization of apoptosis-related molecules is associated with the early therapeutic effects of AHSCT in MS patients. These mechanisms may be involved in the re-establishment of immune tolerance during the first 2 years post-transplantation.

Cell cycle synchronization and analysis of apoptosis-related gene in skin fibroblasts from domestic cat (Felis silvestris catus) and kodkod (Leopardus guigna).

The kodkod population is in constant decrease and the somatic cell nuclear transfer (SCNT) might help to preserve the genetic pool of this species. The cell cycle synchronization of donor cells plays a crucial role in SCNT. The objective of this research was to evaluate two different methods for quiescence induction, serum starvation (SS) and contact inhibition (CI), both for 1, 3 and 5 days, on skin fibroblast from domestic cat and kodkod. Flow cytometry analysis revealed that in domestic cat, SS and CI, both at 3 and 5 days, increased the percentage of fibroblasts in G0/G1 compared to growing cells (GC) (p < .05). In kodkod, only SS for 3 and 5 days and CI for 1 and 3 days increased the percentage of fibroblasts in G0/G1 compared to GC (p < .05). Viability analysis by differential staining revealed that SS for 5 days decreased the proportion of live fibroblasts in domestic cat and kodkod (p < .05). Regarding gene expression analysis, in domestic cat fibroblasts, no differences were found in the BAX/BCL2 ratio in SS and CI (both at 1, 3 and 5 days) compared to GC. In kodkod fibroblasts, BAX/BCL2 ratio was increased in CI at 3 and 5 days compared to SS at 3 and 5 days (p < .05). In conclusion, in kodkod fibroblasts SS for 5 days and CI after 3 days might have a negative impact on cellular viability. According to these results, we suggest SS for 3 days for cell cycle synchronization in kodkod fibroblasts.

Characterization of mesenchymal stem cells in bovine endometrium during follicular phase of oestrous cycle.

Stem cells have been postulated as responsible for cell regeneration in highly and continuously regenerative tissues such as the endometrium. Few studies in cattle have identified and specified the presence of stem cells in the endometrium during the oestrous cycle. The aim of this study was to investigate the presence of mesenchymal stem cells (MSCs) in the bovine endometrium during the follicular phase (FP) of the oestrous cycle. Uterine tissue was collected in the time-frame comprising day 18 of the cycle and ovulation (day 0). We isolated, cultured and expanded four primary cell lines from endometrium and identified byRT-qPCR the expression of OCT4, SOX2 but not NANOG (undifferentiated/embryonic markers), CD44 (MSCs marker) and c-KIT (stem cell marker) genes; and the encoded Oct4, Sox2 and Cd44 proteins by Western blot or immunostaining of paraffin-embedded tissue in endometrium. We demonstrated that cells isolated from bovine endometrium displayed essentially the same gene expression pattern; however, at the protein level, Oct4 and Cd44 were not detected. Besides, they showed typical functional characteristics of MSCs such as fibroblast-like morphology, plastic adherence, high proliferative capacity, clone formation in vitro and the ability to differentiate into chondrogenic, osteogenic and adipogenic lineages. We obtained for the first time an extensive characterization of undifferentiated cells populations contained in the bovine endometrium during the FP of the oestrous cycle.

Effects of meiotic inhibitors and gonadotrophins on porcine oocytes in vitro maturation, fertilization and development.

This study evaluated the effect of three reversible meiotic inhibitors (MINs) and their interaction with gonadotrophins (Gns) on the meiotic maturation and developmental competence of porcine oocytes. In experiment 1, the oocytes were matured for 22 hr in the presence or absence of dbcAMP (1 mM), cycloheximide (7 µM) or cilostamide (20 µM) with or without Gns, and for an additional 22 hr in the absence of MINs and Gns. At 22 hr of maturation, regardless of the presence of Gns, a higher proportion (p < .001) of oocytes cultured in the presence of MINs were effectively arrested at the germinal vesicle stage compared with the oocytes cultured without MINs. At 44 hr of maturation, the proportion of oocytes that reached MII was higher (p < .05) in groups with Gns compared with groups without Gns. In experiment 2, oocytes that were matured as in experiment 1 were inseminated and cultured for 7 days to evaluate fertilization parameters and blastocyst formation. Only oocytes from the dbcAMP + Gns group had higher (p < .05) efficiency of fertilization compared with the other treatment groups. The presence of dbcAMP during maturation also increased (p < .05) blastocyst formation and efficiency of blastocyst formation in both the presence and absence of Gns. These results indicate that the interaction of Gns with the tested MINs improved meiotic progression. In addition, regardless of supplementation with Gns, the presence of dbcAMP during the first maturation period increased and even doubled the capacity of oocytes to develop to the blastocyst stage.

Survival with nonmelanoma skin cancer in Germany.

BACKGROUND: Nonmelanoma skin cancer (NMSC) is the most common cancer in Germany, but detailed information on survival is lacking. OBJECTIVES: To provide survival estimates for female and male patients with basal cell carcinoma (BCC), squamous cell carcinoma (SCC), sarcoma, adenocarcinoma and Merkel cell carcinoma. Further subgroup analyses were carried out by age, tumour stage and body site. METHODS: In total 459 640 patients diagnosed with NMSC in 1997-2011 were included from population-based cancer registers, covering a population of 33 million inhabitants. Age-standardized absolute and relative 5-year and 10-year survival were calculated using period analysis. RESULTS: The absolute and relative 5-year survival were 87·1% and 102·9% for BCC, 77·6% and 93·6% for SCC, 82·1% and 96·0% for sarcoma, 71·4% and 85·7% for adenocarcinoma and 60·0% and 70·7% for Merkel cell carcinoma, respectively. Higher age, female sex and advanced stage were associated with lower survival. CONCLUSIONS: A comprehensive overview of NMSC survival in Germany is provided. The differences between the NMSC subtypes require a more differentiated consideration of patient survival. The survival advantage of patients with BCC may be related to health-promoting factors related to the BCC diagnosis, such as changes to a healthier lifestyle.