Combined Positive Score for Programmed Death Ligand-1 Expression and Inflammatory Microenvironment in Gastrointestinal Stromal Tumors.

Background and Objectives: GISTs are the most frequent type of mesenchymal neoplasm of the digestive tract. The prognosis is mainly determined by tumor dimensions, mitotic rate and location, but other less well-documented factors can influence evolution and survival. The immune microenvironment and checkpoint molecule expression were proven to impact the prognosis in different types of cancer. The aim of this study was to determine PD-L1 expression in GISTs and to evaluate the level of intratumoral immune infiltration in relation to prognostic variables and survival. Materials and Methods: Sixty-five GISTs diagnosed in the same institution between 2015 and 2018 were immunohistochemically tested for PD-L1 and evaluated using CPS. Immune cells were emphasized, with CD3, CD4, CD8, CD20 and CD68 antibodies and quantified. All data were processed using statistical tools. Results: The median age was 61 years (range, 28-78) and 36 patients (55.4%) were males. The location of the tumors was predominantly gastric (46%), followed by the small bowel (17%) and colorectal (6%). In addition, 11% were EGISTs and 20% were secondary tumors (11% metastases and 9% local recurrences). PD-L1 had a variable expression in tumor and inflammatory cells, with a CPS ranging from 0 to 100. Moreover, 64.6% of cases were PD-L1 positive with no significant differences among categories of variables, such as the age and the sex of the patient, tumor location, the primary or secondary character of the tumor, dimensions, mitotic rate, the risk of disease progression and tumor cell type. Immune cells had a variable distribution throughout the tumors. CD3+ lymphocytes were the most frequent type. CD20+ cells were identified in a larger number in tumors ≤5 cm (p = 0.038). PD-L1-positive tumors had a higher number of immune cells, particularly CD3+, CD20+ and CD68+, in comparison to PD-L1-negative ones (p = 0.032, p = 0.051, p = 0.008). Epithelioid and mixed cell-type tumors had a higher number of CD68+ cells. Survival was not influenced by PD-L1 expression; instead, it was decreased in multifocal tumors (p = 0.0001) and in cases with Ki67 ≥ 50% (p = 0.008). Conclusions: PD-L1-positive expression and the presence of different immune cell types, in variable quantities, can contribute to a better understanding of the complex interactions between tumor cells and the microenvironment, with a possible therapeutic role in GISTs.

Profile of common prostate cancer risk variants in an unscreened Romanian population.

To find sequence variants affecting prostate cancer (PCA) susceptibility in an unscreened Romanian population we use a genome-wide association study (GWAS). The study population included 990 unrelated pathologically confirmed PCA cases and 1034 male controls. DNA was genotyped using Illumina SNP arrays, and 24.295.558 variants were imputed using the 1000 Genomes data set. An association test was performed between the imputed markers and PCA. A systematic literature review for variants associated with PCA risk identified 115 unique variants that were tested in the Romanian sample set. Thirty of the previously reported SNPs replicated (P-value < 0.05), with the strongest associations observed at: 8q24.21, 11q13.3, 6q25.3, 5p15.33, 22q13.2, 17q12 and 3q13.2. The replicated variants showing the most significant association in Romania are rs1016343 at 8q24.21 (P = 2.2 × 10-4 ), rs7929962 at 11q13.3 (P = 2.7 × 10-4 ) and rs9364554 at 6q25.2 (P = 4.7 × 10-4 ). None of the variants tested in the Romanian GWAS reached genome-wide significance (P-value <5 × 10-8 ) but 807 markers had P-values <1 × 10-4 . Here, we report the results of the first GWAS of PCA performed in a Romanian population. Our study provides evidence that a substantial fraction of previously validated PCA variants associate with risk in this unscreened Romanian population.

Identification of Lynch syndrome risk variants in the Romanian population.

Two familial forms of colorectal cancer (CRC), Lynch syndrome (LS) and familial adenomatous polyposis (FAP), are caused by rare mutations in DNA mismatch repair genes (MLH1, MSH2, MSH6, PMS2) and the genes APC and MUTYH, respectively. No information is available on the presence of high-risk CRC mutations in the Romanian population. We performed whole-genome sequencing of 61 Romanian CRC cases with a family history of cancer and/or early onset of disease, focusing the analysis on candidate variants in the LS and FAP genes. The frequencies of all candidate variants were assessed in a cohort of 688 CRC cases and 4567 controls. Immunohistochemical (IHC) staining for MLH1, MSH2, MSH6, and PMS2 was performed on tumour tissue. We identified 11 candidate variants in 11 cases; six variants in MLH1, one in MSH6, one in PMS2, and three in APC. Combining information on the predicted impact of the variants on the proteins, IHC results and previous reports, we found three novel pathogenic variants (MLH1:p.Lys84ThrfsTer4, MLH1:p.Ala586CysfsTer7, PMS2:p.Arg211ThrfsTer38), and two novel variants that are unlikely to be pathogenic. Also, we confirmed three previously published pathogenic LS variants and suggest to reclassify a previously reported variant of uncertain significance to pathogenic (MLH1:c.1559-1G>C).

Development, validation, and accuracy of ORCHESTRA emotional exhaustion screening questionnaire among healthcare workers during COVID-19 Pandemic.

BACKGROUND: The COVID-19 pandemic has posed unprecedented challenges, particularly for healthcare workers (HCWs). The prolonged struggles exposed the HCWs to a variety of stressors, potentially leading to burnout. Emotional exhaustion is widely recognized as the core component of burnout. This research aims to conceptualize and develop an emotional exhaustion screening questionnaire through literature review, validation, and accuracy testing. METHOD: A literature review of questionnaires and extraction of items on emotional exhaustion were performed in June 2022. We proceed with the face validity of the items by experts. The items with good content validity ratio and index were selected and reworded to suit the context of HCWs working during the COVID-19 pandemic. A pilot test of the questionnaire was done in the Central University Hospital of Asturias (HUCA) from October to December 2022 with a sample of 148 HCWs from the ORCHESTRA cohort to determine its reliability, convergent validity, and accuracy. RESULTS: Our literature review identified 15 validated questionnaires. After exclusion, 32 items were sent for content validation by experts, yielding five final items that proceeded with the pilot test. Resulting in a Cronbach's alpha-coefficient of .83 for the scale and .78 for dichotomous responses, demonstrating good internal consistency and convergent validity. The result of our accuracy test yielded sensitivity (90.6%) and specificity (91.6%) for the OEEQ scale; and sensitivity (88.7%) and specificity (89.5%) for OEEQ dichotomous responses. CONCLUSION: This study developed and validated the ORCHESTRA Emotional Exhaustion Questionnaire, demonstrating the questionnaire's clarity, relevance, and comprehensibility in screening emotional exhaustion among HCWs.

Incidence and Determinants of Symptomatic and Asymptomatic SARS-CoV-2 Breakthrough Infections After Booster Dose in a Large European Multicentric Cohort of Health Workers-ORCHESTRA Project.

BACKGROUND: SARS-CoV-2 breakthrough infections (BI) after vaccine booster dose are a relevant public health issue. METHODS: Multicentric longitudinal cohort study within the ORCHESTRA project, involving 63,516 health workers (HW) from 14 European settings. The study investigated the cumulative incidence of SARS-CoV-2 BI after booster dose and its correlation with age, sex, job title, previous infection, and time since third dose. RESULTS: 13,093 (20.6%) BI were observed. The cumulative incidence of BI was higher in women and in HW aged < 50 years, but nearly halved after 60 years. Nurses experienced the highest BI incidence, and administrative staff experienced the lowest. The BI incidence was higher in immunosuppressed HW (28.6%) vs others (24.9%). When controlling for gender, age, job title and infection before booster, heterologous vaccination reduced BI incidence with respect to the BNT162b2 mRNA vaccine [Odds Ratio (OR) 0.69, 95% CI 0.63-0.76]. Previous infection protected against asymptomatic infection [Relative Risk Ratio (RRR) of recent infection vs no infection 0.53, 95% CI 0.23-1.20] and even more against symptomatic infections [RRR 0.11, 95% CI 0.05-0.25]. Symptomatic infections increased from 70.5% in HW receiving the booster dose since < 64 days to 86.2% when time elapsed was > 130 days. CONCLUSIONS: The risk of BI after booster is significantly reduced by previous infection, heterologous vaccination, and older ages. Immunosuppression is relevant for increased BI incidence. Time elapsed from booster affects BI severity, confirming the public health usefulness of booster. Further research should focus on BI trend after 4th dose and its relationship with time variables across the epidemics.

PM10 and NO2 Air Pollution and Evolution of COVID-19 Cases in Romania.

Objectives:Based on the correlation between air pollution and COVID-19 incidence/mortality already existing in the literature, we aimed to develop a study to investigate the link between the average level of PM10 (particulate matter 10 - particulate matter 10 microns in diameter) and NO2 (nitrogen dioxide) concentration over five years and the cumulative incidence of COVID-19 cases per 1000 people in Romania. Methods: To assess PM10 and NO2 exposure, we determined the average value of annual PM10 and NO2 concentration for each city over five years (2015-2019). For this purpose, the average of annual PM10 and NO2 concentrations collected from monitoring stations in selected cities was calculated. Then, the annual values over five years were averaged to finally obtain the average PM10 and NO2 concentration for each city. Data on the cumulative number of confirmed cases of COVID-19 up to the 28th of September 2020 were provided by the National Centre for Surveillance and Control of Communicable Diseases (CNSCBT) of the National Institute of Public Health (INSP). The study used the cumulative incidence/hour per 1000 population on 28.09.2020. Results:According to Law no. 104/2011, the annual permissible limit value of PM10 concentration of 40 µg/m³ was not exceeded in any of the 43 cities in our study. The average for all cities was 24.0±4.8 ìg/m³, with a minimum value of average PM10 concentration of 13.9 µg/m3 measured in Alba Iulia and a maximum value of 39.1 µg/m³ in Iasi. The regression model shows that, in Bucharest, 77.9% of the variation in case incidence is explained by the variation in PM10 concentration. In order to find the number of new cases that would correspond to a cumulative incidence of 0.166, taking as an example one of the districts with a population of 259,084, the above regression model shows that an increase in the average PM10 concentration by one unit is associated with 43 new cases. Conclusion:The study demonstrates that an exposure of the population to particulate matter in atmospheric air, at low values, below the permissible limit values but for a long time (the follow-up period in our study was five years, between 2015 and 2019), can have effects on the health status of the population, which becomes much more vulnerable to external agents, in our case pathogenic microorganisms (viruses).

Exhaustion in Healthcare Workers after the First Three Waves of the COVID-19 Pandemic.

This study aims to identify the determinants of exhaustion of frontline and second-line healthcare workers (HCW) during the third wave of the COVID-19 pandemic. A case−control study was conducted based on an anonymously distributed questionnaire, which was completed by 1872 HCW. Exhaustion was assessed with a validated Romanian questionnaire. The Siegrist questionnaire was used to determine workload, reward and overcommitment. Frontline HCW reported significantly more frequent longer working hours (p = 0.0009) and a better perception of the management of the risk for infection (p = 0.0002) than second-line HCW. The effort and overcommitment scores were higher in frontline HCW (9.51 + 1.98 vs. 8.45 + 21, p < 0.001 and 16.34 ± 2.80 vs. 15.24 ± 2.94, p < 0.001, respectively) and the reward scores were lower (5.21 ± 1.522 vs. 5.99 ± 1.44, p < 0.001). In the fully adjusted regression model, age, imbalance between effort and reward, overcommitment and management of the risk of infection in the workplace were associated with the exhaustion score in each category of HCW. The number of working hours was correlated with exhaustion in frontline HCW and occupation in second-line HCW. There were more similarities than differences between frontline and second-line HCW. Even if frontline HCW had a higher risk of exhaustion, the risk was not negligible for all HCW.

Decrease of luminol chemiluminescence upon exposure of human blood serum to 50 Hz electric fields.

The chemiluminescence of luminol, after 1 and 2h in vitro exposure of human serum to 50 Hz electric fields of different intensities, decreases as compared to the controls. This indicates a field-induced decrease in the concentration of the free radicals. The report is limited to the key kinetic and field data, inviting independent kinetic analysis of the data in terms of reaction moments or reaction susceptibilities for the various normal modes indicated by the data.

The effects of low level microwaves on the fluidity of photoreceptor cell membrane.

Due to the extensive use of electromagnetic fields in everyday life, more information is required for the detection of mechanisms of interaction and the possible side effects of electromagnetic radiation on the structure and function of the organism. In this paper, we study the effects of low-power microwaves (2.45 GHz) on the membrane fluidity of rod photoreceptor cells. The retina is expected to be very sensitive to microwave irradiation due to the polar character of the photoreceptor cells [Biochim. Biophys. Acta 1273 (1995) 217] as well as to its high water content [Stud. Biophys. 81 (1981) 39].

Effects of prooxidants on human serum exposed to 50 Hz magnetic fields.

The purpose of this article is to evaluate magnetic field effects (50 Hz, different magnetic intensities) on the chemiluminescence intensity of human serum. We find that 1 and 2 h of exposure increased the chemiluminescence emission. The addition to the serum of prooxidants FeCl(2) and H(2)O(2) in different concentrations increased the chemiluminescence intensity even more.