Differential gene expression of immunity and inflammation genes in colorectal cancer using targeted RNA sequencing.

Introduction: Colorectal cancer (CRC) is a heterogeneous disease caused by molecular changes, as driver mutations, gene methylations, etc., and influenced by tumor microenvironment (TME) pervaded with immune cells with both pro- and anti-tumor effects. The studying of interactions between the immune system (IS) and the TME is important for developing effective immunotherapeutic strategies for CRC. In our study, we focused on the analysis of expression profiles of inflammatory and immune-relevant genes to identify aberrant signaling pathways included in carcinogenesis, metastatic potential of tumors, and association of Kirsten rat sarcoma virus (KRAS) gene mutation. Methods: A total of 91 patients were enrolled in the study. Using NGS, differential gene expression analysis of 11 tumor samples and 11 matching non-tumor controls was carried out by applying a targeted RNA panel for inflammation and immunity genes containing 475 target genes. The obtained data were evaluated by the CLC Genomics Workbench and R library. The significantly differentially expressed genes (DEGs) were analyzed in Reactome GSA software, and some selected DEGs were used for real-time PCR validation. Results: After prioritization, the most significant differences in gene expression were shown by the genes TNFRSF4, IRF7, IL6R, NR3CI, EIF2AK2, MIF, CCL5, TNFSF10, CCL20, CXCL11, RIPK2, and BLNK. Validation analyses on 91 samples showed a correlation between RNA-seq data and qPCR for TNFSF10, RIPK2, and BLNK gene expression. The top differently regulated signaling pathways between the studied groups (cancer vs. control, metastatic vs. primary CRC and KRAS positive and negative CRC) belong to immune system, signal transduction, disease, gene expression, DNA repair, and programmed cell death. Conclusion: Analyzed data suggest the changes at more levels of CRC carcinogenesis, including surface receptors of epithelial or immune cells, its signal transduction pathways, programmed cell death modifications, alterations in DNA repair machinery, and cell cycle control leading to uncontrolled proliferation. This study indicates only basic molecular pathways that enabled the formation of metastatic cancer stem cells and may contribute to clarifying the function of the IS in the TME of CRC. A precise identification of signaling pathways responsible for CRC may help in the selection of personalized pharmacological treatment.

Changes in the prevalence of obesity in Czech adolescents between 2018 and 2022 and its current non-genetic correlates - HBSC study.

BACKGROUND: The main aim of the study is to examine changes in the prevalence of obesity in Czech adolescents between 2018 and 2022 and its current non-genetic correlates with respect to the adolescents' families'socioeconomic status (SES) in 2022. METHODS: The sample of 24,535 adolescents (n = 11,629/12,9062018/2022; boys: 50.4/50.6%2018/2022) aged 10.5-16.5 years that was analysed was drawn from two nationally representative cohorts of Czech youngsters from the last two cycles of the Health Behaviour in School-aged Children (HBSC) online questionnaire survey from 2018 to 2022. Obesity is represented by the > 97th percentile interval on the World Health Organization Body Mass Index percentile chart, with distinctions by sex and the age of adolescents. The differences in the prevalence of obesity between boys and girls from all SES family categories in 2018 and 2022 were tested using a chi-square test (χ2). Multiple logistic regression analysis with repeated measures was used to analyse correlates of obesity in 2022. RESULTS: Between 2018 and 2022, there was no significant difference in the prevalence of obesity in girls or boys in any of the SES categories of families. Adolescents from low-SES families have the highest prevalence of obesity, 11% for boys and 5.8% for girls, significantly higher (p < .001) than its prevalence among adolescents from high-SES families, by + 4.8% points for boys and + 3.9% points for girls. Among adolescents from low-SES families, individuals who engaged in moderate-to-vigorous physical activity (PA) daily (p < .005) or vigorous PA three times per week (p < .05) were significantly less likely to be obese than their less active peers. Skipping breakfast significantly (p < .05) increased the odds of obesity, but only among adolescents from low-SES families. Shorter screen time (ST) significantly (p < .05) reduced the odds of obesity for all categories of adolescent SES. CONCLUSIONS: Obesity is most pronounced in adolescents from low-SES families as a result of a long-term positive energy balance mediated by unbalanced behaviour. Significantly lower odds of obesity in adolescents from low-SES families were confirmed to be associated with regular practice of the recommended PA, shorter ST, and not skipping breakfast.

Modelling Duchenne muscular dystrophy in vitro with newly generated, blood cell-derived induced pluripotent stem cell line ORIONi003-A.

Here, we present newly derived in vitro model for modeling Duchenne muscular dystrophy. Our new cell line was derived by reprogramming of peripheral blood mononuclear cells (isolated from blood from pediatric patient) with Sendai virus encoding Yamanaka factors. Derived iPS cells are capable to differentiate in vitro into three germ layers as verified by immunocytochemistry. When differentiated in special medium, our iPSc formed spontaneously beating cardiomyocytes. As cardiomyopathy is the main clinical complication in patients with Duchenne muscular dystrophy, the cell line bearing the dystrophin gene mutation might be of interest to the research community.

Expression of OCT4 isoforms is reduced in primary colorectal cancer.

Introduction: Colorectal cancer (CRC) is one of the most common types of cancer worldwide. The carcinogenesis of CRC is indeed complex, and there are many different mechanisms and pathways that contribute to the development of malignancy and the progression from primary to metastatic tumors. The OCT4A, encoded by the POU5F1 gene, is a transcription factor responsible for the phenotype of stem cells, maintaining pluripotency and regulation of differentiation. The POU5F1 gene is made up of five exons that can create numerous isoforms through alternative promoter or alternative splicing. In addition to OCT4A, other isoforms called OCT4B are also translated into protein; however, their role in cells has been unclear. The aim of our work was to investigate the expression patterns of OCT4 isoforms in primary and metastatic CRC, providing us with useful information about their role in the development and progression of CRC. Methods: Surgical specimens from a total of 78 patients were collected and isolated from primary tumors (n = 47) and metastases (n = 31). The relative gene expression of OCT4 isoforms was investigated using the RT-qPCR method together with the TaqMan probes for particular OCT4 isoforms. Results: Our results suggest significantly downregulated expression of the OCT4A and OCT4Bs isoforms in both primary (p = 0.0002 and p < 0.0001, respectively) and metastatic tumors (p = 0.0006 and p = 0.00051, respectively) when compared with the control samples. We also observed a correlation between reduced expression of all OCT4 isoforms and both primary and left-sided tumors (p = 0.001 and p = 0.030, respectively). On the other hand, the expression of all OCT4 isoforms was significantly upregulated in metastases compared with primary tumors (p < 0.0001). Discussion: Unlike previous reports, we found out that the expression of OCT4A, OCT4Bs, and all OCT4 isoforms was significantly reduced in primary tumors and metastases compared with control samples. On the other hand, we supposed that the expression rate of all OCT4 isoforms may be related to the cancer type and side, as well as to liver metastases. However, further studies are required to investigate the detailed expression patterns and significance of individual OCT4 isoforms in carcinogenesis.

Measurement Invariance of the WHO-5 Well-Being Index: Evidence from 15 European Countries.

(1) Background: The World Health Organization (WHO)-5 Well-Being Index has been used in many epidemiological studies to assess adolescent mental well-being. However, cross-country comparisons of this instrument among adolescents are scarce and, so far, no good-fitting, common invariant measurement model across countries has been reported. The present study aims to evaluate and establish a version of the WHO-5 Well-Being Index that allows for a valid cross-country comparison of adolescent self-reported mental well-being. (2) Methods: Using data from the 2018 Health Behaviour in School-aged Children study, we evaluated the measurement model and measurement invariance of the five items of the WHO-5 Well-Being Index. We used nationally representative samples of 11-, 13-, and 15-year-old adolescents (N = 74,071) from fifteen countries and regions in Europe. Measurement invariance of the WHO-5 was assessed using a series (country, gender, and age) of multi-group confirmatory factor analyses. In addition, we evaluated the convergent validity of the measure by testing its correlations with psychosomatic complaints, life satisfaction, and self-rated health. (3) Results: We found that WHO-5 does not show good psychometric properties or good measurement invariance fit. However, by excluding the first item of the scale ("I have felt cheerful and in good spirits"), the WHO-4, consisting of the other four original items, had good psychometric properties, and demonstrated good suitability for cross-national comparisons (as well as age and gender) in adolescent mental well-being. (4) Conclusions: The present study introduces the WHO-4-a revised version of the WHO-5-, that allows for a valid comparison of mental well-being across fifteen countries and regions in Europe. The WHO-4 proved to be a reliable and valid instrument to assess mental well-being in the adolescent population.

In vitro modeling of amyotrophic lateral sclerosis with induced pluripotent stem cell technology-derived cell line ORIONi002-A.

We present here a new iPS cell line for modeling sporadic form of ALS. Cell line was generated by reprogramming skin fibroblasts isolated with explant culture technology from skin biopsy, donated by ALS patient. For reprogramming, polycistronic self-replicating RNA vector was used and derived iPS cells were characterized by immunocytochemistry and FACS (pluripotent factors expression), karyotyping, STR fingerprinting analysis and in vitro differentiation assay. New cell line showed normal (46, XY) karyotype and differentiated in vitro into cells from three germ layers. STR analysis proved the origin and originality of the cell line.

Cancer stem cell marker expression and methylation status in patients with colorectal cancer.

The number of individuals diagnosed with colorectal cancer (CRC) has been on an alarming upward trajectory over the past decade. In some countries, this cancer represents one of the most frequently diagnosed types of neoplasia. Therefore, it is an important demand to study the pathology underlying this disease to gain insights into the mechanism of resistance to treatment. Resistance of tumors to chemotherapy and tumor aggressiveness have been associated with a minor population of neoplastic cells, which are considered to be responsible for tumor recurrence. These types of neoplastic cells are known as cancer stem cells, which have been previously reported to serve an important role in pathogenesis of this malignant disease. Slovakia has one of the highest incidence rates of CRC worldwide. In the present study, the aim was to classify the abundance of selected stem cell markers (CD133, CD166 and Lgr5) in CRC tumors using flow cytometry. In addition, the methylation status of selected genomic regions of CRC biomarkers (ADAMTS16, MGMT, PROM1 (CD133), LGR5 and ALCAM) was investigated by pyrosequencing in a cohort of patients from Martin University Hospital, Martin, Slovakia. Samples from both primary tumors and metastatic tumors were tested. Analysis of DNA methylation in the genomic regions of indicated five CRC biomarkers was also performed, which revealed the highest levels of methylation in the A disintegrin and metalloproteinase with thrombospondin motifs 16 and O6-methyguanine-DNA methyl transferase genes, whereas the lowest levels of methylation were found in genes expressing prominin-1, leucine-rich repeat-containing G-protein-coupled receptor 5 and activated leukocyte cell adhesion molecule. Furthermore, tumor tissues from metastases showed significantly higher levels of CD133+ cells compared with that in primary tumors. Higher levels of CD133+ cells correlated with TNM stage and the invasiveness of CRC into the lymphatic system. Although relatively small number of samples was processed, CD133 marker was consider to be important marker in pathology of CRC.

Long-term trends in the consumption of sugary and diet soft drinks among adolescents: a cross-national survey in 21 European countries.

PURPOSE: To assess country-level trends in the prevalence of daily consumption of sugary (2002-2018) and diet (2006-2018) soft drinks among European adolescents, overall and by family material affluence. METHODS: We used 2002, 2006, 2010, 2014 and 2018 data from the 'Health Behaviour in School-aged Children' survey. Nationally representative samples of adolescents completed a standardised questionnaire at school, including a short food frequency questionnaire (n = 530,976 and 21 countries for sugary soft drinks; n = 61,487 and 4 countries for diet soft drinks). We classified adolescents into three socioeconomic categories for each country and survey year, using the Family Affluence Scale. Multilevel logistic models estimated time trends, by country. RESULTS: Sugary soft drinks: the prevalence of daily consumption (≥ 1×/day) declined in 21/21 countries (Plinear trends ≤ 0.002). Absolute [range - 31.7 to - 3.4% points] and relative [range - 84.8 to - 22.3%] reductions varied considerably across countries, with the largest declines in Ireland, England and Norway. In 3/21 countries, the prevalence of daily consumption decreased more strongly in the most affluent adolescents than in the least affluent ones (P ≤ 0.002). Daily consumption was more prevalent among the least affluent adolescents in 11/21 countries in 2018 (P ≤ 0.002). Diet soft drinks: overall, daily consumption decreased over time in 4/4 countries (Plinear trends ≤ 0.002), more largely among the most affluent adolescents in 1/4 country (P ≤ 0.002). CONCLUSIONS: Daily consumption of sugary and diet soft drinks in European adolescents decreased between 2002 (2006 for diet drinks) and 2018. Public health interventions should continue discouraging daily soft drink consumption, particularly among adolescents from lower socioeconomic groups.

Establishment of PANDA - a new human pancreatic ductal adenocarcinoma cell line with 3D cell culture technology.

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive type of malignancy with one of the worst prognoses amongst any type of cancer. Surgery is applicable only to the limited number of patients with locally resectable tumors and currently represents the only curative treatment option. Treatment with chemotherapy and radiotherapy can only extend patient survival. Despite advances in conventional therapies, the five-year survival of PDAC remained largely unchanged. New in vitro and in vivo models are therefore urgently needed to investigate this type of cancer. Here, we present the establishment and characterization of a novel pancreatic cancer cell line, isolated from a patient with PDAC. Cell line abbreviated as PANDA (PANncreatic Ductal Adenocarcinoma) was established with an optimized 3D culture protocol published previously by our group. The new cancer cell line "PANDA" represents a novel in vitro approach for PDAC cancer research and new therapy testing.

Laparoscopic Right Hemicolectomy for Appendiceal Mucocele.

Appendiceal mucocele is a rare disease with an incidence of 0.07-0.63% of all appendectomies and was first described in 1842 by Carl von Rokitansky. It is defined as an abnormal intraluminal accumulation of mucin. The clinical picture of AM can vary from asymptomatic mass in the right lower quadrant to symptoms of acute appendicitis. In some cases, AM can be found accidentally on CT performed due to other reasons or during surgery. Diagnosis consists mainly of imaging methods such as ultrasound, CT, and MRI with the finding of encapsulated cystic mass with calcifications. The main goal of surgical treatment is to remove an intact mucocele and prevent spillage of mucin into the peritoneal cavity. We present a case of large mucocele treated with laparoscopic right hemicolectomy.

Screening for the Key Proteins Associated with Rete Testis Invasion in Clinical Stage I Seminoma via Label-Free Quantitative Mass Spectrometry.

Rete testis invasion (RTI) is an unfavourable prognostic factor for the risk of relapse in clinical stage I (CS I) seminoma patients. Notably, no evidence of difference in the proteome of RTI-positive vs. -negative CS I seminomas has been reported yet. Here, a quantitative proteomic approach was used to investigate RTI-associated proteins. 64 proteins were differentially expressed in RTI-positive compared to -negative CS I seminomas. Of them, 14-3-3γ, ezrin, filamin A, Parkinsonism-associated deglycase 7 (PARK7), vimentin and vinculin, were validated in CS I seminoma patient cohort. As shown by multivariate analysis controlling for clinical confounders, PARK7 and filamin A expression lowered the risk of RTI, while 14-3-3γ expression increased it. Therefore, we suggest that in real clinical biopsy specimens, the expression level of these proteins may reflect prognosis in CS I seminoma patients.

Detection of therapeutically relevant and concomitant rare somatic variants in colorectal cancer.

In colorectal cancer (CRC), clinically relevant biomarkers are known for genome-guided therapy that can be detected by both first and next generation methods. The aim of our work was to introduce a robust NGS assay that will be able to detect, in addition to standard predictive single nucleotide-based biomarkers, even rare and concomitant clinically relevant variants. Another aim was to identify truncating mutations in APC and pathogenic variants in TP53 to divide patients into potentially prognostic groups. A multigene panel with hotspots in 50 cancer-critical genes was used. Finally, 86 patients diagnosed with primary or metastatic colorectal cancer were enrolled. In total, there were identified 163 pathogenic variants, among them in the genes most recurrent mutated in CRC such as TP53 (49%), the RAS family genes KRAS and NRAS (47%), APC (43%), and PIK3CA (15%). In 30 samples, two driver mutations were present in one sample, 11 patients were without any mutations covered by this panel. In one patient, a novel variant in BRAF p.D594E was found, not previously seen in CRC, and was concomitant with KRAS p.G12A. In KRAS, a potentially sensitive mutation to anti-EGFR therapy p.A59T was found along with the PIK3CA missense variant p.E545K. It was possible to divide patients into groups based on the occurrence of truncating APC variant alone or concomitant with TP53 or KRAS. Our results demonstrate the potential of small multigene panels that can be used in diagnostics for the detection of rare therapeutically relevant variants. Moreover, the division of patients into groups based on the presence of APC and TP53 mutations enables this panel to be used in retrospective studies on the effectiveness of treatment with anti-EGFR inhibitors.

Cross-national time trends in adolescent alcohol use from 2002 to 2014.

BACKGROUND: Adolescent alcohol consumption is a major public health concern that should be continuously monitored. This study aims (i) to analyze country-level trends in weekly alcohol consumption, drunkenness and early initiation in alcohol consumption and drunkenness among 15-year-old adolescents from 39 countries and regions across Europe and North America between 2002 and 2014 and (ii) to examine the geographical patterns in adolescent alcohol-related behaviours. METHODS: The sample was composed of 250 161 adolescents aged 15 from 39 countries and regions from Europe and North America. Survey years were 2002, 2006, 2010 and 2014. The alcohol consumption and drunkenness items of the HBSC questionnaire were employed. Prevalence ratios and 95% confidence intervals were estimated using Poisson regression models with robust variance. RESULTS: Data show a general decrease in all four alcohol variables between 2002 and 2014 except for some countries. However, there is variability both within a country (depending on the alcohol-related behaviour under study) and across countries (in the beginning and shape of trends). Some countries have not reduced or even increased their levels in some variables. Although some particularities have persisted over time, there are no robust patterns by regions. CONCLUSIONS: Despite an overall decrease in adolescent alcohol consumption, special attention should be paid to those countries where declines are not present, or despite decreasing, rates are still high. Further research is needed to clarify factors associated with adolescent drinking, to better understand country specificities and to implement effective policies.

Time trends in consumption of sugar-sweetened beverages and related socioeconomic differences among adolescents in Eastern Europe: signs of a nutrition transition?

BACKGROUND: High intake of sugar-sweetened beverages (SSBs) contributes to detrimental cardio-metabolic indicators in youth. Monitoring of SSB consumption is lacking in Eastern Europe. OBJECTIVES: We assessed trends in the prevalence of adolescent daily consumption of SSBs in 14 Eastern European countries between 2002 and 2018, both overall and according to family material affluence. METHODS: We used 2002, 2006, 2010, 2014, and 2018 data of the Health Behaviour in School-Aged Children school-based study (repeated cross-sectional). Nationally representative samples of adolescents aged 11, 13, and 15 years were included (n = 325,184; 51.2% girls). Adolescents completed a standardized questionnaire, including a question on SSB consumption frequency. We categorized adolescents into 3 socioeconomic groups based on the relative Family Affluence Scale (FAS). Adjusted prevalences of daily SSB consumption by survey year, as well as country-level time trends between 2002 and 2018, were computed using multilevel logistic models (overall and by FAS groups). RESULTS: In 2018, the prevalence of adolescents consuming SSBs every day varied considerably between countries (range, 5.1%-28.1%). Between 2002 and 2018, the prevalence of daily SSB consumption declined in 10/14 countries (P for linear trends ≤ 0.004). The largest reductions were observed in Slovenia (OR, 0.48; 95% CI: 0.45-0.50) and the Russian Federation (OR, 0.67; 95% CI: 0.64-0.70). Daily SSB consumption was reduced at faster rates among the most affluent adolescents (who were larger consumers in 2002) than in the least affluent adolescents in 11/14 countries (P for linear trends ≤ 0.004). Thus, differences between FAS groups narrowed over time or even reversed, leading to larger proportions of daily consumers in the least affluent adolescents in 2018 in 5/14 countries (P ≤ 0.05). CONCLUSIONS: Adolescent daily consumption of SSBs decreased between 2002 and 2018 in most Eastern European countries. Declines were larger among higher-affluence adolescents. These results are useful to evaluate and plan interventions promoting healthy childhood diets.

Genomic profile and immune contexture in colorectal cancer-relevance for prognosis and immunotherapy.

Colorectal cancer (CRC) is one of the leading cancers in both genders. TNM staging system is still the most commonly used tumor classification and prognostic system. The disadvantage of TNM is that the prognostic information it provides is incomplete, and patients with the same histological tumor stages may differ significantly in the clinical outcome. Therefore, the identification of new prognostic parameters is crucial. The carcinogenic process that gives rise to an individual tumor is unique and tumor microenviroment should be taken into consideration. In CRC, T-cell infiltration is not homogenous, and recent studies are mostly focusing on memory T-cells and CD8 cells in predicting disease-free survival (DFS) and overall survival (OS). It seems that DFS and OS are not only dependent on microsatellite instable or stable status but mostly on the levels of expression of the immune signatures. Also, patients with high infiltration of cytotoxic and memory cells have significantly better outcome. This review consolidates current knowledge and recent research about importance of immune-cell-associated proteins, specific gene profiles of immune cells and immunotherapy in CRC. We also discussed cell-specific signatures in cancer treatment.

Time trends in adolescent mental wellbeing in the Czech Republic between 2002 and 2018: gender, age and socioeconomic differences.

OBJECTIVES: Recent literature indicates a decline over time in adolescent mental wellbeing but results are inconsistent and rely mainly on data from Western societies. This study investigates time trends in adolescent mental wellbeing (psychological and somatic complaints, life satisfaction) among Czech adolescents and explores the moderating role of gender, age and socioeconomic status. METHODS: Nationally representative data from 29,376 Czech adolescents (50.8% girls, mean age = 13.43; SD = 1.65) across five Health Behaviour in School-aged Children (HBSC) surveys (2002, 2006, 2010, 2014, 2018) were used. Hierarchical regression models estimated national trends in adolescent mental wellbeing and established the moderating role of gender, age and socioeconomic status. RESULTS: From 2002 to 2018, an increase in the psychological complaints was observed. Life satisfaction decreased over time up to 2014 only, whereas somatic symptoms increased until 2010, followed by a decline in 2014 and 2018. Girls, older adolescents and those from low family affluence reported poorer mental wellbeing. Gender gap increased over time for psychological complaints and life satisfaction. Socioeconomic inequalities gap remained stable over the investigated timeframe. CONCLUSIONS: Our findings do not provide evidence for substantial temporal changes in mental wellbeing among adolescents in the Czech Republic. Yet, the increase in psychological complaints has been consistent which is an indicator of a small decline over time in adolescent mental wellbeing. Furthermore, the gender gap in mental wellbeing increased over time, whereas the age and socioeconomic differences remained relatively stable. This calls for the attention of public health professionals and policy makers from the Czech Republic.

Multi-center real-world comparison of the fully automated Idylla™ microsatellite instability assay with routine molecular methods and immunohistochemistry on formalin-fixed paraffin-embedded tissue of colorectal cancer.

Microsatellite instability (MSI) is present in 15-20% of primary colorectal cancers. MSI status is assessed to detect Lynch syndrome, guide adjuvant chemotherapy, determine prognosis, and use as a companion test for checkpoint blockade inhibitors. Traditionally, MSI status is determined by immunohistochemistry or molecular methods. The Idylla™ MSI Assay is a fully automated molecular method (including automated result interpretation), using seven novel MSI biomarkers (ACVR2A, BTBD7, DIDO1, MRE11, RYR3, SEC31A, SULF2) and not requiring matched normal tissue. In this real-world global study, 44 clinical centers performed Idylla™ testing on a total of 1301 archived colorectal cancer formalin-fixed, paraffin-embedded (FFPE) tissue sections and compared Idylla™ results against available results from routine diagnostic testing in those sites. MSI mutations detected with the Idylla™ MSI Assay were equally distributed over the seven biomarkers, and 84.48% of the MSI-high samples had ≥ 5 mutated biomarkers, while 98.25% of the microsatellite-stable samples had zero mutated biomarkers. The concordance level between the Idylla™ MSI Assay and immunohistochemistry was 96.39% (988/1025); 17/37 discordant samples were found to be concordant when a third method was used. Compared with routine molecular methods, the concordance level was 98.01% (789/805); third-method analysis found concordance for 8/16 discordant samples. The failure rate of the Idylla™ MSI Assay (0.23%; 3/1301) was lower than that of referenced immunohistochemistry (4.37%; 47/1075) or molecular assays (0.86%; 7/812). In conclusion, lower failure rates and high concordance levels were found between the Idylla™ MSI Assay and routine tests.

A Psychometric Evaluation of the Guilt and Shame Experience Scale (GSES) on a Representative Adolescent Sample: A Low Differentiation between Guilt and Shame.

The Guilt and Shame Experience Scale (GSES) is a new, brief self-report instrument for assessing experiences of guilt and shame. It includes two distinct scales: feelings of shame and feelings of guilt. The present report focuses on results from a final validation study using a nationally representative sample of 7899 adolescents (M age = 14.5 ± 1.1 years, 50.7% boys) who participated in the 2014 Health Behavior in School-aged Children study. For factor analysis, the dataset was divided into two groups. One group (n = 3950) was used for the Exploratory Factor Analysis (EFA) and the second (n = 3949) for the Confirmatory Factor Analysis (CFA). The EFA results in a one-factor model of the GSES scale, while the CFA suggests a two-factor solution mirroring two scales, feelings of shame and feelings of guilt. Both models have a good fit to the data, and the scale also showed high internal consistency (Cronbach's alpha = 0.89). A nonparametric comparison of different sociodemographic groups showed a higher disposition for experiencing guilt and shame among girls, students of the ninth grade, and religious respondents. A comparison of the results to previously published results obtained from adults indicates that adolescence is a developmental period involving low differentiation between moral emotions like guilt and shame compared with adulthood. Moreover, positive association with religious attendance shows a need of addressing these issues in a pastoral care setting.

Insulinoma presenting with postprandial hypoglycemia and a low body mass index: A case report.

BACKGROUND: Insulinomas are the most common type of functioning endocrine neoplasms of the pancreas presenting hypoglycemic symptoms. Patients characteristically develop symptoms while fasting, but some patients have reported symptoms only in the postprandial state. Repeated and prolonged hypoglycemic episodes can reduce the awareness of adrenergic symptoms, and patients may have amnesia, which delays diagnosis. CASE SUMMARY: We describe a case of a 24-year-old underweight patient who showed hypoglycemic symptoms for almost 6 years. Although patients with insulinoma characteristically develop symptoms while fasting, this young man had hypoglycemic symptoms up to one hour postprandially, especially after high-sugar meals and after physical activity. The fasting tests and imaging methods performed at local hospitals were evaluated as negative for abnormal results. However, brown adipose tissue exhibited increased metabolic activity, and some muscle groups had histological changes as indicated by positron emission tomography with 2-deoxy-2-[fluorine-18]fluoro-D-glucose integrated with computed tomography. Glycogen deficiency was also histologically confirmed. The patient's symptoms progressed over the years and occurred more frequently, i.e., several times a month, and the patient had reduced awareness of adrenergic symptoms. The follow-up fasting test was positive, and the imaging results showed a tumor in the head of the pancreas. The patient underwent laparotomy with enucleation of the insulinoma. CONCLUSION: Weight gain and fasting hypoglycemia are not necessarily characteristics of insulinoma. In prolonged cases, adrenergic symptoms can be suppressed.

Pathway Analysis of Selected Circulating miRNAs in Plasma of Breast Cancer Patients: A Preliminary Study.

MicroRNAs in the circulation of breast cancer (BC) patients have great potential for the early diagnosis, treatment and monitoring of breast cancer. The aim of this preliminary study was to obtain the expression profile of selected miRNAs in the plasma of BC patients that could discriminate BC patients from healthy volunteers and may be useful in early detection of BC. Significantly deregulated miRNAs were evaluated by pathway analysis with the prediction of potential miRNA targets. The study enrolled plasma samples from 65 BC patients and 34 healthy volunteers. Selected miRNAs were screened in pilot testing by the real-time PCR (qPCR) method, and the most appropriate reference genes were selected for normalisation by the geNorm algorithm. In the final testing, we detected miR-99a, miR-130a, miR-484 and miR-1260a (p < 0.05) as significantly up-regulated in the plasma of BC patients. Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway analysis revealed that all significantly deregulated miRNAs are involved in the Hippo and Transforming Growth Factor-beta (TGF-beta) signalling pathways. Our study confirmed a different profile of selected circulating miRNAs in the plasma of BC patients with an emphasis on some critical points in the analysis process.