Sex-specific cardiovascular remodeling leads to a divergent sex-dependent development of heart failure in aged hypertensive rats.

INTRODUCTION: The prevalence of heart failure with preserved ejection fraction (HFpEF) is continuously rising and predominantly affects older women often hypertensive and/or obese or diabetic. Indeed, there is evidence on sex differences in the development of HF. Hence, we studied cardiovascular performance dependent on sex and age as well as pathomechanisms on a cellular and molecular level. METHODS: We studied 15-week- and 1-year-old female and male hypertensive transgenic rats carrying the mouse Ren-2 renin gene (TG) and compared them to wild-type (WT) controls at the same age. We tracked blood pressure and cardiac function via echocardiography. After sacrificing the 1-year survivors we studied vascular smooth muscle and endothelial function. Isolated single skinned cardiomyocytes were used to determine passive stiffness and Ca2+-dependent force. In addition, Western blots were applied to analyse the phosphorylation status of sarcomeric regulatory proteins, titin and of protein kinases AMPK, PKG, CaMKII as well as their expression. Protein kinase activity assays were used to measure activities of CaMKII, PKG and angiotensin-converting enzyme (ACE). RESULTS: TG male rats showed significantly higher mortality at 1 year than females or WT male rats. Left ventricular (LV) ejection fraction was specifically reduced in male, but not in female TG rats, while LV diastolic dysfunction was evident in both TG sexes, but LV hypertrophy, increased LV ACE activity, and reduced AMPK activity as evident from AMPK hypophosphorylation were specific to male rats. Sex differences were also observed in vascular and cardiomyocyte function showing different response to acetylcholine and Ca2+-sensitivity of force production, respectively cardiomyocyte functional changes were associated with altered phosphorylation states of cardiac myosin binding protein C and cardiac troponin I phosphorylation in TG males only. Cardiomyocyte passive stiffness was increased in TG animals. On a molecular level titin phosphorylation pattern was altered, though alterations were sex-specific. Thus, also the reduction of PKG expression and activity was more pronounced in TG females. However, cardiomyocyte passive stiffness was restored by PKG and CaMKII treatments in both TG sexes. CONCLUSION: Here we demonstrated divergent sex-specific cardiovascular adaptation to the over-activation of the renin-angiotensin system in the rat. Higher mortality of male TG rats in contrast to female TG rats was observed as well as reduced LV systolic function, whereas females mainly developed HFpEF. Though both sexes developed increased myocardial stiffness to which an impaired titin function contributes to a sex-specific molecular mechanism. The functional derangements of titin are due to a sex-specific divergent regulation of PKG and CaMKII systems.

[External radiation therapy of primary central nervous system tumors - current trends and practice]. / A primer központi idegrendszeri tumorok konvencionális külso sugárkezelése ­ aktuális trendek és gyakorlat.

The family of primary central nervous system (CNS) tumors is a highly heterogeneous group of diseases. In the complex care of CNS tumors, in addition to surgical and systemic treatments, modern external beam radiation therapy (EBRT) plays a prominent role. This summary article provides an overview of the current indications related to EBRT, diagnostic procedures, contouring, planning, and radiation therapy techniques and applications that can be used in daily routine for adults' most common primary CNS tumors.

Stereotactic body radiotherapy in lung cancer: a contemporary review.

The treatment of early stage non-small cell lung cancer (NSCLC) has improved enormously in the last two decades. Although surgery is not the only choice, lobectomy is still the gold standard treatment type for operable patients. For inoperable patients stereotactic body radiotherapy (SBRT) should be offered, reaching very high local control and overall survival rates. With SBRT we can precisely irradiate small, well-defined lesions with high doses. To select the appropriate fractionation schedule it is important to determine the size, localization and extent of the lung tumor. The introduction of novel and further developed planning (contouring guidelines, diagnostic image application, planning systems) and delivery techniques (motion management, image guided radiotherapy) led to lower rates of side effects and more conformal target volume coverage. The purpose of this study is to summarize the current developments, randomised studies, guidelines about lung SBRT, with emphasis on the possibility of increasing local control and overall rates in "fit," operable patients as well, so SBRT would be eligible in place of surgery.

LINAC-based SBRT in treating early-stage NSCLC patients-single institution experience and survival data analysis.

Aim: This single institute prospective study aimed to evaluate the feasibility of LINAC-based stereotactic body radiotherapy (SBRT) in treating patients with early-stage non-small cell lung cancer (NSLSC). We focused on the survival data with the local and distant control profiles and the cancer- and non-cancer-specific survival. Treatment-related side effects were also collected and analyzed. Methods: Patients with early-stage NSCLC between January 2018 and October 2021 were included in our prospective study; a total of 77 patients receiving LINAC-based SBRT were analyzed. All patients had pretreatment multidisciplinary tumor board decisions on SBRT. The average patient age was 68.8 years (median: 70 years, range: 52-82); 70 patients were in ECOG 0 status (91%), while seven patients were in ECOG 1-2 status (9%). 52% of the patients (40) had histologically verified NSCLC, and the other 48% were verified based on PETCT results. We applied the SBRT scheme 8 x 7.5 Gy for central tumors (74%) or 4 x 12 Gy for peripheral tumors (26%). Results: The mean follow-up time was 25.4 months (median 23, range 18-50). The Kaplan-Meier estimation for overall survival in patients receiving LINAC-based SBRT was 41.67 months. Of the 77 patients treated by SBRT, death was reported for 17 patients (9 cases cancer-specific, 8 cases non-cancer specific reason). The mean local tumor control was 34.25 months (range 8.4-41), and the mean systemic control was 24.24 months (range 7-25). During the treatments, no Grade I-II were reported; in 30 cases, Grade I non-symptomatic treatment-related lung fibrosis and two asymptomatic rib fractures were reported. Conclusion: In the treatment of early-stage NSCLC, LINAC-based SBRT can be a feasible alternative to surgery. Although we reported worse OS data in our patient cohort compared to the literature, the higher older average age and the initial worse general condition (ECOG1-2) in our patient cohort appear to be the reason for this difference. With the comparable local control and survival data and the favorable side effect profile, SBRT might be preferable over surgery in selected cases.

Occupational Stress Levels among Radiologists and Radiographers in Hungary during the COVID-19 Era.

The COVID-19 pandemic has heightened stress levels, potentially affecting the occupational wellbeing of radiographers and radiologists. Our study aimed to assess occupational stress levels within the radiology department and identify contributing factors. A cross-sectional survey was conducted between September and November 2022, with participants comprising radiographers and radiologists affiliated with the Hungarian Society of Radiographers and the Hungarian Society of Radiologists. The online survey collected socio-demographic and COVID-19 data, and the participants completed an effort-reward imbalance questionnaire. The analysis of 406 responses revealed significantly higher effort-reward imbalance (ERI) levels among the radiologists compared to the radiographers (p < 0.05). The healthcare professionals with over 30 years of experience exhibited significantly lower ERI levels than those with 1-9 years, 10-19 years, or 20-29 years of experience (p < 0.05). Additionally, the individuals aged 31-40 demonstrated higher ERI levels compared to their counterparts aged 19-30, 41-50, and over 51 (p < 0.05). The respondents cohabiting with a spouse/partner reported significantly higher stress levels than their single colleagues (p < 0.05), while the dog owners exhibited significantly lower ERI levels (p < 0.05). Elevated occupational stress highlights specific groups requiring targeted interventions to reduce stress and mitigate burnout among radiologists and radiographers.

Reduced burnout in medical and health science students during the pandemic COVID-19 - a follow-up study of a single institution in Hungary.

BACKGROUND: The coronavirus pandemic has significantly impacted lives worldwide, especially of medical and health science students. In Hungary, education has been relegated to the online space, with a substantial proportion of students having to attend medical secondments. Increased stress, uncertainty, and the presence of medical secondments can have an impact on students' premature burnout. METHODS: In 2021, we conducted a follow-up survey among students of the University of Pécs studying medicine and health sciences in two data collection periods (from March to May and September to November). Our online questionnaire consisted of the Maslach Burnout Inventory General Survey for Students and our self-designed questionnaire. We used descriptive and paired two-sample t-tests for data analysis at a 95% confidence interval (p ≤ 0.05). RESULTS: We excluded from our survey respondents whose data we could not follow-up; finally, 183 students' responses were analyzed. The majority of students were female (n = 148; 80.9%). Overall, there was a significant decrease in both exhaustion (EX) and cynicism (CY) scores (p = 0.001; p = 0.004). Female respondents had higher EX scores, but a significant decrease was observed for both genders (p ≤ 0.05). Excluding paramedic students, a significant decrease in EX scores was observed for the specialties we studied (p ≤ 0.05). General medicine students' CY scores decreased; physiotherapy students' profesisonal efficacy (PE) scores increased significantly (p ≤ 0.05). Students who were on medical secondments (n = 127; 69. 4%) were found to be more affected by burnout, but in all cases, these scores significantly improved (p ≤ 0.05). Students serving in the National Ambulance Service (n = 76; 41.5%), Hospitals (n = 44; 24.0%), or both (n = 7; 3.8%) had a significant decrease in their burnout score (p ≤ 0.05). Students who served in either a hospital or a hospital and National Ambulance Service had significantly improved CY and PE scores (p ≤ 0.05). Students concerned about their health had elevated EX and CY scores, which also improved (p ≤ 0.05). CONCLUSIONS: In conclusion, medical secondments positively affected student burnout scores for medicine and health sciences students at our institution. This fact implies that it is necessary to have more internships in real-life settings during the training. TRIAL REGISTRATION: Our survey has been approved by the Medical Research Council (Case No IV/4573-1/2021/ECU).

Gene Therapy of Extracellular Vesicles in Cardiovascular and Metabolic Diseases.

The ultimate and most complex form of treating human diseases is embodied by gene therapy. For an effective gene therapeutic product we need to hack the cellular plasma membrane entry-system, then escaping degradation in the cytosol and in most cases, we need an efficient hacking of the nuclear membrane-system, achieving the delivery of genetic construct into the central stage of the target cells: nucleoplasm or chromosomal DNA found in this highly controlled space. These steps need to be performed in a targeted, ordered, and efficient way. Possessing intrinsic ability of nucleic acid and protein delivery, extracellular vesicles can bypass biological barriers and may be able to deliver a next-generation platform for gene therapy. Fine-tuned genetic constructs included in (synthetic) extracellular vesicles may provide an upgraded approach to the current gene therapeutical technologies by significantly upgrading and improving biosafety, versatility, and delivery, thus evoking the desired therapeutic response. This chapter addresses the main types, vectors, challenges, and safety issues of gene therapy. Afterwards, a brief introduction and beneficial roles of extracellular vesicles are given. The concept of engineering vesicles for gene therapy is also discussed. A snapshot of most relevant clinical trials in the field of cardiovascular and metabolic diseases is shown. Finally, a wrap-up and outlook about gene therapy are presented.

Oxidative stress and inflammation distinctly drive molecular mechanisms of diastolic dysfunction and remodeling in female and male heart failure with preserved ejection fraction rats.

Heart failure with preserved ejection fraction (HFpEF) is a complex cardiovascular insufficiency syndrome presenting with an ejection fraction (EF) of greater than 50% along with different proinflammatory and metabolic co-morbidities. Despite previous work provided key insights into our understanding of HFpEF, effective treatments are still limited. In the current study we attempted to unravel the molecular basis of sex-dependent differences in HFpEF pathology. We analyzed left ventricular samples from 1-year-old female and male transgenic (TG) rats homozygous for the rat Ren-2 renin gene (mRen2) characterized with hypertension and diastolic dysfunction and compared it to age-matched female and male wild type rats (WT) served as control. Cardiomyocytes from female and male TG rats exhibited an elevated titin-based stiffness (Fpassive), which was corrected to control level upon treatment with reduced glutathione indicating titin oxidation. This was accompanied with high levels of oxidative stress in TG rats with more prominent effects in female group. In vitro supplementation with heat shock proteins (HSPs) reversed the elevated Fpassive indicating restoration of their cytoprotective function. Furthermore, the TG group exhibited high levels of proinflammatory cytokines with significant alterations in apoptotic and autophagy pathways in both sexes. Distinct alterations in the expression of several proteins between both sexes suggest their differential impact on disease development and necessitate distinct treatment options. Hence, our data suggested that oxidative stress and inflammation distinctly drive diastolic dysfunction and remodeling in female and male rats with HFpEF and that the sex-dependent mechanisms contribute to HF pathology.

Burnout and occupational stress among Hungarian radiographers working in emergency and non-emergency departments during COVID-19 pandemic.

INTRODUCTION: The increased workload caused by the coronavirus pandemic may have had a significant impact on the mental health of radiographers. The aim of our study was to investigate burnout and occupational stress in radiographers working in emergency departments (ED) and non-emergency departments (NED). METHODS: Quantitative, cross-sectional, descriptive research was carried out among radiographers working in the public health sector in Hungary. Due to the cross-sectional nature of our survey, there was no overlap between the ED and NED groups. For data collection, we used simultaneously the Maslach Burnout Inventory (MBI), the Effort-Reward Imbalance questionnaire (ERI), and our self-designed questionnaire. RESULTS: We excluded incomplete questionnaires from our survey; finally, 439 responses were evaluated. Significantly higher scores for depersonalisation (DP; 8.43 (SD = 6.69) vs. 5.63 (SD = 4.21) and emotional exhaustion (EE; 25.07 (SD = 11.41) vs. 19.72 (SD = 11.72)) were observed in radiographers working in ED (p = 0.001; p = 0.001) when compared to NED. Male radiographers working in ED aged 20-29 and 30-39 years with experience of 1-9 years were more affected by DP (p ≤ 0.05). Worrying about one's own health had a negative effect on DP and EE (p ≤ 0.05). Having close friend with a COVID-19 infection had a negative effect on EE (p ≤ 0.05); not being infected with coronavirus, not being quarantined and relocating within the workplace had a positive effect on personal accomplishment (PA); radiographers who were 50 years or older with 20-29 years of experience were more affected by depersonalisation (DP); and those who worried about their health had significantly higher stress scores (p ≤ 0.05) in both ED and NED settings. CONCLUSION: Male radiographers at the beginning of their careers were more affected by burnout. Employment in EDs had a negative impact on DP and EE. IMPLICATIONS FOR PRACTICE: Our results support the implementation of interventions to counter the effects of occupational stress and burnout among radiographers working in ED.

Upregulation of exofacial peroxiredoxin-thioredoxin system of lymphocytes and monocytes in preeclampsia.

OBJECTIVES: An imbalanced redox homeostasis resulting in oxidative stress is present in preeclampsia. Peroxiredoxin-1 (PRDX1) and thioredoxin-1 (TRX1) regulatory enzymes are also contributing to the redox homeostasis, but were not investigated so far in preeclampsia. Thus, we have aimed to characterize PRDX1, TRX1 and oxidative stress biomarkers in blood samples of pregnant women with preeclampsia. STUDY DESIGN: Twelve patients with preeclampsia (PE) were enrolled into the study. Seven third trimester healthy pregnant women (HP) were accepted as control group. MAIN OUTCOME MEASURES: Peripheral venous blood samples of healthy and preeclamptic pregnant women were analyzed. Plasma level of advanced oxidation protein products (AOPP) was determined by spectrophotometry. The exofacial PRDX1 and TRX1 expression of lymphocytes and monocytes was detected by flow cytometry. RESULTS: The plasma AOPP level was significantly higher in preeclampsia compared to the healthy pregnant group. Significantly higher percentage of PRDX1 and TRX1 expressing lymphocytes and monocytes were detected in the blood samples of preeclamptic women compared to healthy pregnant controls. The ratio of circulating PRDX1 and TRX1 expressing lymphocytes and monocytes showed a significant inverse correlation with the birth weight of newborns. CONCLUSIONS: We have revealed that the level of advanced oxidation protein products is increased and the exofacial peroxiredoxin-1 and thioredoxin-1 system in lymphocytes and monocytes is upregulated in preeclampsia. In addition, the ratio of peroxiredoxin-1 and thioredoxin-1 positive circulating lymphocytes and monocytes correlates inversely with the neonatal birth weight, which finding indicates that pregnancies complicated by intrauterine growth restriction are accompanied by a higher level of oxidative stress.

Optical genome mapping and revisiting short-read genome sequencing data reveal previously overlooked structural variants disrupting retinal disease-associated genes.

PURPOSE: Structural variants (SVs) play an important role in inherited retinal diseases (IRD). Although the identification of SVs significantly improved upon the availability of genome sequencing, it is expected that involvement of SVs in IRDs is higher than anticipated. We revisited short-read genome sequencing data to enhance the identification of gene-disruptive SVs. METHODS: Optical genome mapping was performed to improve SV detection in short-read genome sequencing-negative cases. In addition, reanalysis of short-read genome sequencing data was performed to improve the interpretation of SVs and to re-establish SV prioritization criteria. RESULTS: In a monoallelic USH2A case, optical genome mapping identified a pericentric inversion (173 megabase), with 1 breakpoint disrupting USH2A. Retrospectively, the variant could be observed in genome sequencing data but was previously deemed false positive. Reanalysis of short-read genome sequencing data (427 IRD cases) was performed which yielded 30 pathogenic SVs affecting, among other genes, USH2A (n = 15), PRPF31 (n = 3), and EYS (n = 2). Eight of these (>25%) were overlooked during previous analyses. CONCLUSION: Critical evaluation of our findings allowed us to re-establish and improve our SV prioritization and interpretation guidelines, which will prevent missing pathogenic events in future analyses. Our data suggest that more attention should be paid to SV interpretation and the current contribution of SVs in IRDs is still underestimated.

Sleep Quality and Perceived Stress among Health Science Students during Online Education-A Single Institution Study.

Recently, online education has been gaining prominence in university life. Our survey aimed to examine sleep quality and perceived stress levels among students at the University of Pécs Faculty of Health Sciences. A cross-sectional, quantitative, descriptive survey was conducted between February and March 2023. The online survey included the Hungarian versions of the internationally validated Athens Insomnia Scale (AIS) and Perceived Stress Scale (PSS). Statistical analysis involved descriptive statistics, independent t-tests, analysis of variance (ANOVA), and Mann-Whitney and Kruskal-Wallis tests (p < 0.05). We analyzed 304 responses, and females dominated (n = 270; 88.8%). Students in a relationship had significantly higher AIS scores (t = -2.470; p = 0.014). Medium average (2.50-3.49) students and those who rarely/never exercise showed significantly higher AIS and PSS (p ≤ 0.05). Students on the phone/watching a series during online education, daily laptop/TV use for more than 2 h, and pre-sleep use of smart devices for more than 60 min also negatively affected AIS and PSS scores (p ≤ 0.05). Nursing, physiotherapy, and radiography students were the most affected regarding insomnia and perceived stress (p ≤ 0.05). Our survey shows that excessive smart device use and lack of exercise are associated with higher stress levels and poorer sleep quality.

Enhancing Molecular Testing for Effective Delivery of Actionable Gene Diagnostics.

There is a deep need to navigate within our genomic data to find, understand and pave the way for disease-specific treatments, as the clinical diagnostic journey provides only limited guidance. The human genome is enclosed in every nucleated cell, and yet at the single-cell resolution many unanswered questions remain, as most of the sequencing techniques use a bulk approach. Therefore, heterogeneity, mosaicism and many complex structural variants remain partially uncovered. As a conceptual approach, nanopore-based sequencing holds the promise of being a single-molecule-based, long-read and high-resolution technique, with the ability of uncovering the nucleic acid sequence and methylation almost in real time. A key limiting factor of current clinical genetics is the deciphering of key disease-causing genomic sequences. As the technological revolution is expanding regarding genetic data, the interpretation of genotype-phenotype correlations should be made with fine caution, as more and more evidence points toward the presence of more than one pathogenic variant acting together as a result of intergenic interplay in the background of a certain phenotype observed in a patient. This is in conjunction with the observation that many inheritable disorders manifest in a phenotypic spectrum, even in an intra-familial way. In the present review, we summarized the relevant data on nanopore sequencing regarding clinical genomics as well as highlighted the importance and content of pre-test and post-test genetic counselling, yielding a complex approach to phenotype-driven molecular diagnosis. This should significantly lower the time-to-right diagnosis as well lower the time required to complete a currently incomplete genotype-phenotype axis, which will boost the chance of establishing a new actionable diagnosis followed by therapeutical approach.

Altered Cellular Protein Quality Control System Modulates Cardiomyocyte Function in Volume Overload-Induced Hypertrophy.

Volume-induced hypertrophy is one of the risk factors for cardiac morbidity and mortality. In addition, mechanical and metabolic dysfunction, aging, and cellular redox balance are also contributing factors to the disease progression. In this study, we used volume overload (VO), which was induced by an aortocaval fistula in 2-month-old male Wistar rats, and sham-operated animals served as control. Functional parameters were measured by transthoracic echocardiography at termination 4- or 8-months after VO. The animals showed hypertrophic remodeling that was accompanied by mechanical dysfunction and increased cardiomyocyte stiffness. These alterations were reversible upon treatment with glutathione. Cardiomyocyte dysfunction was associated with elevated oxidative stress markers with unchanged inflammatory signaling pathways. In addition, we observed altered phosphorylation status of small heat shock proteins 27 and 70 and diminished protease expression caspases 3 compared to the matched control group, indicating an impaired protein quality control system. Such alterations might be attributed to the increased oxidative stress as anticipated from the enhanced titin oxidation, ubiquitination, and the elevation in oxidative stress markers. Our study showed an early pathological response to VO, which manifests in cardiomyocyte mechanical dysfunction and dysregulated signaling pathways associated with enhanced oxidative stress and an impaired protein quality control system.

Diagnostics and follow-up strategy for Silver­Russell syndrome based on a case report showing familial accumulation / A Silver­Russell-szindróma diagnosztikai lépései és terápiás lehetoségei egy családi halmozódást mutató eset kapcsán

"Characterized by both intrauterine and postnatal growth retardation, and consequent small stature, Silver­Russell syndrome is associated with typical minor anomalies (relative macrocephalia, protruding forehead, downturned corners of mouth, micrognathia, low set ears, facial, skeletal and limb asymmetry) and findings involving mainly the endocrine system. The molecular background of the syndrome is complex, but it is most often caused by the involvement of critical regions of chromosome 11 and/or chromosome 7. Beside the molecular diagnosis, the Netchine­Harbison clinical scoring system aims to contribute to the successful diagnosis of Silver­Russell syndrome. Although Silver­Russell syndrome is mostly sporadic, in our case report we present an extremely rare familial accumulation, where three of four siblings are affected by Silver­Russell syndrome. Early diagnosis is important to initiate adequate feeding and nutritional guidance, enhance early development and start growth hormone therapy as soon as possible. We would like to emphasize that management and long-term follow-up is crucial to prevent potential complications and treat specific issues appropriately."

Current Understanding of Molecular Pathophysiology of Heart Failure With Preserved Ejection Fraction.

Heart Failure (HF) is the most common cause of hospitalization in the Western societies. HF is a heterogeneous and complex syndrome that may result from any dysfunction of systolic or diastolic capacity. Abnormal diastolic left ventricular function with impaired relaxation and increased diastolic stiffness is characteristic of heart failure with preserved ejection fraction (HFpEF). HFpEF accounts for more than 50% of all cases of HF. The prevalence increases with age: from around 1% for those aged <55 years to >10% in those aged 70 years or over. Nearly 50% of HF patients have HFrEF and the other 50% have HFpEF/HFmrEF, mainly based on studies in hospitalized patients. The ESC Long-Term Registry, in the outpatient setting, reports that 60% have HFrEF, 24% have HFmrEF, and 16% have HFpEF. To some extent, more than 50% of HF patients are female. HFpEF is closely associated with co-morbidities, age, and gender. Epidemiological evidence suggests that HFpEF is highly represented in older obese women and proposed as 'obese female HFpEF phenotype'. While HFrEF phenotype is more a male phenotype. In addition, metabolic abnormalities and hemodynamic perturbations in obese HFpEF patients appear to have a greater impact in women then in men (Sorimachi et al., European J of Heart Fail, 2022, 22). To date, numerous clinical trials of HFpEF treatments have produced disappointing results. This outcome suggests that a "one size fits all" approach to HFpEF may be inappropriate and supports the use of tailored, personalized therapeutic strategies with specific treatments for distinct HFpEF phenotypes. The most important mediators of diastolic stiffness are the cardiomyocytes, endothelial cells, and extracellular matrix (ECM). The complex physiological signal transduction networks that respond to the dual challenges of inflammatory and oxidative stress are major factors that promote the development of HFpEF pathologies. These signalling networks contribute to the development of the diseases. Inhibition and/or attenuation of these signalling networks also delays the onset of disease. In this review, we discuss the molecular mechanisms associated with the physiological responses to inflammation and oxidative stress and emphasize the nature of the contribution of most important cells to the development of HFpEF via increased inflammation and oxidative stress.

SARS-CoV-2 infects human cardiomyocytes promoted by inflammation and oxidative stress.

INTRODUCTION: The respiratory illness triggered by severe acute respiratory syndrome virus-2 (SARS-CoV-2) is often particularly serious or fatal amongst patients with pre-existing heart conditions. Although the mechanisms underlying SARS-CoV-2-related cardiac damage remain elusive, inflammation (i.e. 'cytokine storm') and oxidative stress are likely involved. METHODS AND RESULTS: Here we sought to determine: 1) if cardiomyocytes are targeted by SARS-CoV-2 and 2) how inflammation and oxidative stress promote the viral entry into cardiac cells. We analysed pro-inflammatory and oxidative stress and its impact on virus entry and virus-associated cardiac damage from SARS-CoV-2 infected patients and compared it to left ventricular myocardial tissues obtained from non-infected transplanted hearts either from end stage heart failure or non-failing hearts (donor group). We found that neuropilin-1 potentiates SARS-CoV-2 entry into human cardiomyocytes, a phenomenon driven by inflammatory and oxidant signals. These changes accounted for increased proteases activity and apoptotic markers thus leading to cell damage and apoptosis. CONCLUSION: This study provides new insights into the mechanisms of SARS-CoV-2 entry into the heart and defines promising targets for antiviral interventions for COVID-19 patients with pre-existing heart conditions or patients with co-morbidities.

Ca2+/calmodulin-dependent protein kinase II and protein kinase G oxidation contributes to impaired sarcomeric proteins in hypertrophy model.

AIMS: Volume overload (VO) induced hypertrophy is one of the hallmarks to the development of heart diseases. Understanding the compensatory mechanisms involved in this process might help preventing the disease progression. METHODS AND RESULTS: Therefore, the present study used 2 months old Wistar rats, which underwent an aortocaval fistula to develop VO-induced hypertrophy. The animals were subdivided into four different groups, two sham operated animals served as age-matched controls and two groups with aortocaval fistula. Echocardiography was performed prior termination after 4- and 8-month. Functional and molecular changes of several sarcomeric proteins and their signalling pathways involved in the regulation and modulation of cardiomyocyte function were investigated. RESULTS: The model was characterized with preserved ejection fraction in all groups and with elevated heart/body weight ratio, left/right ventricular and atrial weight at 4- and 8-month, which indicates VO-induced hypertrophy. In addition, 8-months groups showed increased left ventricular internal diameter during diastole, RV internal diameter, stroke volume and velocity-time index compared with their age-matched controls. These changes were accompanied by increased Ca2+ sensitivity and titin-based cardiomyocyte stiffness in 8-month VO rats compared with other groups. The altered cardiomyocyte mechanics was associated with phosphorylation deficit of sarcomeric proteins cardiac troponin I, myosin binding protein C and titin, also accompanied with impaired signalling pathways involved in phosphorylation of these sarcomeric proteins in 8-month VO rats compared with age-matched control group. Impaired protein phosphorylation status and dysregulated signalling pathways were associated with significant alterations in the oxidative status of both kinases CaMKII and PKG explaining by this the elevated Ca2+ sensitivity and titin-based cardiomyocyte stiffness and perhaps the development of hypertrophy. CONCLUSIONS: Our findings showed VO-induced cardiomyocyte dysfunction via deranged phosphorylation of myofilament proteins and signalling pathways due to increased oxidative state of CaMKII and PKG and this might contribute to the development of hypertrophy.

Plan Quality Assessment of Fractionated Stereotactic Radiotherapy Treatment Plans in Patients With Brain Metastases.

Background and Purpose: The treatment options available in the management of brain metastases includes fractionated stereotactic radiotherapy (FSRT) and stereotactic radiosurgery (SRS) treatments. FSRT treatments have proved to be useful mainly in the treatment of larger volumes. This study aims to evaluate the FSRT treatment technique used in our department based on various plan quality indices. Methods and Materials: 24 treatment plans of 23 patients were analyzed. Volumetric modulated arc therapy (VMAT) plans were generated in line with the department protocol. The following parameters were extracted: Radiation Therapy Oncology Group conformity index (RTOG CI), Paddick conformity index (Paddick CI), gradient index (GI), quality index (Q), homogeneity index (HI), and V24.4 volume as a parallel index of V12 used at SRS plan evaluation. Results: Plan conformity was acceptable, RTOG CI mean was 0.942; Paddick CI mean was 0.824. The mean GI value was 6.146. The mean of HI and Q indices were 1.263 and 0.94, respectively. V24.4 mean was 33.434 cm3. All plans achieved clinically acceptable organs-at-risk (OAR) constraints. PTV volumes were clustered into either 10 cm3 or 15 cm3 bins depending on the plan quality metric we used. The mean values show a balanced distribution of plan indices along the various PTV bins. Discussion: Our results based on the derived indices show that our FSRT approach can achieve clinically acceptable treatment plans. Furthermore, the clustering of PTV volumes show that these plan quality metrics remain acceptable for a wide spectrum of PTV volumes.