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Nitric oxide synthases (NOSs) and Na+/K+-ATPase are enzymes essential for regular functioning of the heart. Since both enzymes are under insulin and androgen regulation and since insulin action and androgen level were disturbed in polycystic ovary syndrome (PCOS), we hypothesized that cardiac nitric oxide (NO) production and sodium/potassium transport would be deteriorated in PCOS. To test our hypothesis we introduced animal model of PCOS based on dihydrotestosterone (DHT) treatment of female Wistar rats and analyzed protein expression, phosphorylation or subcellular localization of endothelial NOS (eNOS), inducible NOS (iNOS) and alpha subunits of Na+/K+-ATPase in the heart. Obtained results indicate that DHT treatment significantly decreased cardiac eNOS protein level and activating phosphorylation at serine 1 177, while inhibitory phosphorylation at threonine 495 was increased. In contrast to expression of eNOS, iNOS protein level in the heart of DHT-treated rats was significantly elevated. Furthermore, cardiac protein level of alpha 1 subunit of the ATPase, as well as its plasma membrane content, were decreased in rats with PCOS. In line with this, alpha 2 subunit protein level in fraction of plasma membranes was also significantly below control level. In conclusion, DHT treatment impaired effectiveness of NOSs and Na+/K+-ATPase in the female rat heart. Regarding the importance of NO production and sodium/potassium transport in the cardiac contraction and blood flow regulation, it implicates strong consequences of PCOS for heart functioning. [ABSTRACT FROM AUTHOR]

Molecular mechanisms underlying pathophysiology of polycystic ovary syndrome (PCOS), especially those related to cortisol signaling, are poorly understood. We hypothesized that modulation of glucocorticoid receptor (GR) expression and function, may underlie possible PCOS-related impairment of feedback inhibition of hypothalamic-pituitary-adrenocortical (HPA) axis activity and thus contribute to increased adrenal androgen production in women with PCOS.24 normal-weight and 31 obese women with PCOS were compared to 25 normal-weight controls. Fasting blood samples were collected for measurements of serum concentrations of dehydroepiandrosterone sulfate, testosterone, sex hormone-binding globulin, insulin, basal cortisol and cortisol after oral administration of 0.5 mg dexamethasone. Concentrations of GR mRNA, GR protein, mineralocorticoid receptor (MR) protein and heat shock proteins (Hsps), as well as the number of GR per cell (Bmax) and its equilibrium dissociation constant (KD) were measured in isolated peripheral blood mononuclear cells.An increase in HPA axis sensitivity to dexamethasone, an elevation of the GR protein concentration, and unaltered receptor functional status were found in both normal-weight and obese women with PCOS vs. healthy controls. Lymphocyte MR, Hsp90 and Hsp70 concentrations, and MR/GR ratio were similar in all groups. Correlation between Bmax and KD was weaker in the group of obese women with PCOS than in the other 2 groups.The results did not confirm the initial hypothesis, but imply that PCOS is associated with increased GR protein concentration and HPA axis sensitivity to dexamethasone. [ABSTRACT FROM AUTHOR]

Accessible summary We examined effects of physical activity on fitness of children with and without hyperactivity., No significant differences were observed in fitness levels between two groups., Hyperactive children improved coordination of whole body, trunk strength and agility., Abstract The objectives of this study were to assess the effects of participating in regular physical activity on the fitness of children with hyperactivity. The study compared a sample of children who were assessed as hyperactive with the same number of children rated as non-hyperactive. The Conners' Rating Scale was used for hyperactivity evaluation, while fitness levels were assessed using a battery of six motor tests prior to and following the completion of the physical exercise programme. The findings indicated that while overall fitness levels of both groups improved, the differences were not statistically significant. However, improvements shown within the group of hyperactive children with regard to coordination of the whole body, trunk strength and agility were statistically significant. Physical activity may be associated with enhanced levels of some aspects of physical fitness. [ABSTRACT FROM AUTHOR]

This paper analyses the influence of mechanical loads on railway vehicle solid wheel reliability. Reliability calculations is carried out according to the specifically set methodology — reliability is calculated according to the data about working loads and critical stress distribution. The influences on solid wheel reliability that are analysed are the following: standard deviation of lateral force distribution and mathematical expectation and standard deviation of the vertical force. [ABSTRACT FROM AUTHOR]

Endocrine disruptors (EDs) are considered to have an impact on the function of reproductive axis at different levels as well on reproductive organs in both sexes. Complexity of female reproductive system influenced with various stressors including EDs lead to morphological and functional alterations. This is resulting in modulation of neuroendocrine regulation with consequent developmental irregularities and derangements, causative infertility, endometriosis as well as premature ovarian insufficiency or polycystic ovary syndrome. A number of experimental clues was obtained on female animal models using various EDs such as synthetic estrogens and phytoestrogens, neurotransmitters, pesticides or various chemicals. These substances lead towards consequent derangement of the neuroendocrine control of reproduction from early phases of reproductive development towards different phases of adult reproductive period. This text will address some novel insights into the effects of EDs on neuroendocrine regulation of gonadal axis, effects on ovaries as well on endometrium during implantation period. [ABSTRACT FROM AUTHOR]

The aim of the study was to investigate the morphology of mesothelial cells of the peritoneum of patients with terminal renal failure (TRF), taken by the biopsy immediately before the onset of peritonal dialysis (PD), and to compare it with the findings in patients with PD. The samples were prepared in the way standard for light microscopy and transmission electron microscopy. In patients with TRF intracytoplasmatic inclusions could be observed, unusual protrusions of mesothelial apical surfaces, deformation of mesothelial cells and their detachment from the basal membrane, as well as the dilatated cisternae of granulated endoplasmatic reticulum with filamentous structures in some of them. In patients on PD cytoplasmic protrusions of different shapes and contents were observed at the surface of mesothelial cells, multiplication of basal membrane, occurrence of young forms of mesothelial cells as well as the detachment of those cells from the basal lamina. [ABSTRACT FROM AUTHOR]

Background/objectives: Multiple sclerosis is characterised by the manifestation of heterogeneous symptoms that affect daily functioning. Patients face physical impairment, psychological problems and socioeconomic changes. Depending on the form of the disease, they may have different needs, which are often unsatisfied and could be overcome by including palliative care in the treatment. Despite the above, not enough is known about this population's needs for palliative care. This cross-sectional study aimed to identify the need for palliative care and assess the functional dependence level in daily living activities in patients with multiple sclerosis., Methods: The sample consisted of 120 patients from the Neurology Clinic of the University Clinical Center of Vojvodina, Serbia. The following research instruments were used: a General questionnaire, Sheffield Profile for Assessment and Referral for Care, Multiple Sclerosis Impact Scale and Barthel Index., Results: The need for palliative care was expressed by 36.7% of patients, who expressed the most concern for the physical symptoms, independence and activity domains. Also, it was determined that the disease prevents patients from doing demanding physical tasks and that they are worried about multiple sclerosis. Almost a quarter of patients have had complete/severe dependence on other persons in performing daily activities., Conclusions: The most frequently reported physical symptoms by patients with MS were weakness, bladder problems, fatigue and different levels of dependence, while concerns related to sexual health, low mood and anxiety emerged as key challenges in the psychological domain. These findings underscore the importance of conducting needs assessments to guide the development of an appropriate palliative care model for MS patients.

As healthcare workers run a high and constant occupational risk of hepatitis B virus (HBV) infection through exposure to biological material, vaccination is mandatory as well as the monitoring of antibody levels one to two months after complete immunisation. The aim of this descriptive cross-sectional study was to determine HBV vaccine coverage of 200 primary and secondary healthcare workers (100 each) from Šabac, Serbia and their blood anti-HBs titre. We also wanted to identify factors that could predict the titre. Anti-HBV vaccination covered all participants, of whom 89.5 % were fully vaccinated, and 85 % had a protective antibody titre. We found a statistically significant association between antibody titre and the number of received vaccine doses, chronic jaundice, autoimmune disease, and cancer in our participants. The fact that 15 % did not achieve the protective antibody titre confirms the necessity of its control after immunisation, which is not routinely carried out in most countries, Serbia included. It is, therefore, necessary to develop a detailed strategy for monitoring vaccination and serological status of healthcare workers in order to improve their safety at work. An important role should also be given to continuous education of healthcare workers from the beginning of schooling to the end of their professional career., (© 2024 Slobodanka Bogdanović Vasić et al., published by Sciendo.)

Background: To provide competent and respectful sexual health care, a critical and comprehensive assessment of students' attitudes towards sexuality is needed, which implies using reliable and valid questionnaires., Aim: To assess whether the extended version of the Students Attitude Towards Addressing Sexual Health (SA-SH-Ext) provides different, additional information about students' attitudes towards providing sexual healthcare compared to the Sexual Attitude and Beliefs Survey (SABS), to explore SA-SH-Ext potential response patterns through latent class analysis and to assess students' attitudes towards providing sexual healthcare and evaluate the differences in their attitudes to the cultural background and sociodemographic characteristics., Design: Multicentre, descriptive, analytical, comparative, and correlational cross-sectional study at three nursing faculties., Participants: Nursing students n = 514 (Serbia n = 180, Poland n = 150 and Lithuania n = 184)., Methods: A general questionnaire for obtaining sociodemographic data, SA-SH-Ext and SABS were used as students' report measures., Results: One part of the variance of each instrument does not overlap with the other, indicating that these instruments, in addition to the shared variance, provide different, additional information. A solution with three latent classes was found in the domain of responses to the SA-SH-Ext items. The response pattern placed nursing students in the class Comfortable and prepared in some situations, and the SABS score revealed students' moderately positive attitudes towards providing sexual healthcare. Significantly more conservative attitudes were demonstrated by students from Poland, those identifying their religious affiliation as Catholic and females, while first-year students showed more positive attitudes than those in later years of study., Conclusions: The SA-SH-Ext and SABS enable a reliable and quick assessment of nursing students' comfort and competence in providing sexual healthcare. However, the differences in students' attitudes towards sexuality concerning belonging to a specific group imply a need for curriculum redesign and facilitating students to be more open to communication about sexuality with people with mental illnesses and intellectual and physical disabilities., Competing Interests: Conflict of interest The authors declare that there is no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Polycystic ovary syndrome (PCOS) is the most common endocrine disorder in women of reproductive age and is closely associated with chronic low-grade inflammation and insulin resistance. To clarify the contribution of prepubertal weight gain to the development of insulin resistance in PCOS, we investigated the effects of early postnatal overfeeding on inflammatory and energy-sensing pathways as well as on markers of insulin signaling in the liver of the PCOS rat model., Methods: Obesity induced by overfeeding was achieved by reducing litter size, while the PCOS-like condition was developed by treatment with 5α-dihydrotestosterone (DHT). Western blot and qPCR were used to analyze the expression of pro-inflammatory transcription factors and cytokines, as well as markers of the energy sensing and insulin signaling pathways., Results: The results showed that hepatic insulin sensitivity was impaired only in DHT-treated rats raised in small litters, as evidenced by increased phosphorylation of IRS1 on Ser307 and decreased expression of total IRS1. Postnatal overfeeding stimulated JNK1 activation independent of hyperandrogenemia; nevertheless, the synergistic effect of both factors triggered NLRP3 activation and increased IL1β expression in the small litter DHT-treated group. This pro-inflammatory state was accompanied by decreased activatory phosphorylation of AMPK and reduced levels of its protein targets., Conclusions: Overfeeding in the early postnatal period leads to a decrease in hepatic insulin sensitivity in the rat model of PCOS, which is associated with decreased activation of AMPK and stimulation of the hepatic NLRP3-IL1β signaling pathway. Accordingly, the inhibition of NLRP3 activation could provide a basis for the development of new therapeutic strategies for the treatment of insulin resistance in women with PCOS., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Veličković, Mićić, Teofilović, Milovanovic, Jovanović, Djordjevic, Macut and Vojnović Milutinović.)

Obesity is a pressing problem worldwide for which standard therapeutic strategies have limited effectiveness. The use of natural products seems to be a promising approach to alleviate obesity and its associated complications. The tepals of Crocus sativus (Cr) plant, usually wasted in saffron production, are an unexplored source of bioactive compounds. Our aim was to elucidate the mechanisms of Cr tepals extract in obesity by investigating its effects on adipocyte differentiation, visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) hypertrophy, and lipid metabolism in an animal model of diet-induced obesity. To this end, mouse 3T3-F442A preadipocytes were treated with Cr tepals extract and the expression of adipocyte differentiation genes was determined. Caloric intake, body mass, triglycerides, systemic insulin sensitivity, histology, insulin signaling, and lipid metabolism in VAT and SAT were analyzed in mice fed a 60% fat diet for 14 weeks and treated orally with Cr tepals extract during the last 5 weeks of the diet. We demonstrated for the first time that Cr tepals extract inhibits adipocyte differentiation in vitro. The animal model confirmed that oral treatment with Cr tepals extract results in weight loss, improved systemic insulin sensitivity, lower triglycerides, and improved lipid peroxidation. The suppressive effect of Cr tepals extract on adipocyte hypertrophy and inflammation was observed only in SAT, which, together with preserved SAT insulin signaling, most likely contributed to improved systemic insulin sensitivity. Our results suggest the functionality of SAT as a possible target for the treatment of obesity and its complications., (© 2024 International Union of Biochemistry and Molecular Biology.)

Increased fructose consumption and chronic stress, the major characteristics of modern lifestyle, impact human health; however, the consequences of their combination on the uterus remain understudied. In this study, we investigated contractile activity, morphology, and intracellular activity of antioxidant enzymes in uteri from virgin Wistar rats subjected to liquid fructose supplementation and/or unpredictable stress over 9 weeks. Contractile activity and uterine response to oxytocin or adrenaline were examined ex vivo using isolated bath chambers. Fructose supplementation, irrespective of stress, affected uterine morphology by increasing endometrium while decreasing myometrium volume density, attenuated uterine response to increasing doses of oxytocin, and increased glutathione peroxidase activity. Stress, irrespective of fructose, attenuated dose-dependent adrenaline-induced uterine relaxation. Stress, when applied solely, decreased mitochondrial superoxide dismutase activity. In the combined treatment, irregular estrous cycles and both reduced response to oxytocin and to adrenaline (as a consequence of fructose consumption and exposure to stress), along with fructose-related alteration of uterine morphology, were detected. In conclusion, fructose and stress affect uterine contractile activity, irrespective of each other, by inducing completely distinct responses in isolated uteri. In the combined treatment, the effects of both factors were evident, suggesting that the combination exerts more detrimental effects on the uterus than each factor individually.

Many patients require administering one or more blood components during hospitalisation in the Intensive Care Unit (ICU). Therefore, nurses' knowledge of who is responsible for immediately administering blood transfusions, monitoring patients, and identifying and managing transfusion reactions is crucial. This cross-sectional descriptive-analytical study aimed to assess the knowledge of ICU nurses in tertiary healthcare institutions about blood transfusion procedures. The questionnaire about the transfusion procedure was designed and reviewed by experts. The questionnaire consisted of 29 items divided into three domains. The scores on the knowledge test ranged from 10 to 27. Generally, 57.7% of nurses had moderate, 23.4% low, and 18.9% high levels of knowledge about the transfusion procedure. Most nurses answered correctly about refreezing fresh frozen plasma, verifying the transfusion product, and identifying the patient. Of the nurses, 91.0% would recognise mild allergic reactions, and 98.2% knew about the supervision of sedated patients. Nurses showed poor knowledge of the length of usage of the same transfusion system for red blood cells, labelling, and transfusion administration in febrile patients. Nurses with higher education and longer working experience had significantly better outcomes ( p = 0.000) on the knowledge test. Continuous education of ICU nurses on safe transfusion usage is recommended.

Purpose : To translate the original Jefferson Scale of Empathy-Health Profession Students version in a group of nursing students and asses its psychometrical characteristics according to the COnsensus-based Standards for the selection of health Measurement INstruments guidelines. Design : A descriptive cross-sectional study. Methods : The study included 345 bachelor's and master's students in the nursing degree program. Construct validity was tested by exploratory factor analysis using principal component analysis with varimax rotation. Convergent validity was tested using the Pearson correlation coefficient to test the relationship between empathy and emotional intelligence. The reliability of internal consistency was estimated using Cronbach's alpha. Findings : The final version of the 16-item scale with its three-factor structure has a high reliability with a Cronbach's alpha value of 0.86, and the Pearson correlation coefficient revealed a positive and significant relationship between emotional intelligence and the total score of the empathy scale and the three subscales. Conclusion : The contribution of the study is significant as the instrument is qualitatively validated and its content is clinically and educationally important for understanding the concept of empathy, especially in the education of nursing students. The correct assessment of empathy contributes to a better understanding and implementation of the components of holistic care., Competing Interests: Ethical ConsiderationThis study received approval from the Higher Education Institution committee (IRB approval number: 2158/97-97-07-20-03). The study adhered to the principles of the Helsinki Declaration of Human Rights, with participation being entirely voluntary and respondents informed of their right to withdraw from the study without consequences. The responses were anonymized, ensuring the confidentiality of the students' identities. During the test-retest phase, an anonymized and easily memorable code was used. Each questionnaire was accompanied by informed consent and consent for voluntary participation in the research. Consent for the translation and validation of the JSE-HPS was obtained from the Thomas Jefferson University (Center for Research in Medical Education and Health Care), and consent for the use of the ESCQ-45 was obtained from author Vladimir Takšić

Introduction: Effective diabetes self-management and collaborative responsibility sharing with parents are imperative for pediatric patients with type 1 diabetes mellitus, particularly as they gradually assume more self-care responsibilities. The primary goal of this study was to assess differences in adherence to self-care activities regarding sociodemographics and clinical characteristics in pediatric patients with type 1 diabetes. The secondary goal of this study was to understand the level of parental involvement in diabetes management and to assess the pediatric patients' behaviors (independent or dependent on disease self-management) that relate to sociodemographic and clinical characteristics., Methods: This was a comparative cross-sectional and correlational study. The study sample included 182 children and adolescents who had been diagnosed with type 1 diabetes at least 3 months prior. Data collection instruments included a sociodemographic and questionnaire about Adherence to self-care activities and parental involvement in diabetes self-management, as well as a documentation sheet for recording clinical data., Results: A majority of participants (71%) exhibited non-adherence to self-care tasks, despite 78.0% asserting their independence in diabetes self-management. Notably, insufficient parental involvement in administering insulin therapy significantly predicted severe hypoglycemic episodes., Conclusions: Pediatric patients dealing with type 1 diabetes demonstrate a substantial degree of autonomy in managing their condition, paradoxically coupled with self-reported non-adherence to critical self-care responsibilities. Notably, children (aged 8-12) rely more heavily on parental support, especially concerning insulin therapy administration. The study underscores the crucial role of parental engagement in insulin therapy, as its deficiency significantly predicts the likelihood of severe hypoglycemic episodes., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Smudja et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Objective: The number of participants in sports or some form of recreation globally has led to an increase in the incidence of anterior cruciate ligament (ACL) injuries and the number of surgeries performed. Although it does not belong to risky surgical interventions, this operation is accompanied by complications that slow down post-operative rehabilitation. The objective is to analyze the effects of intra-articular (IA) injection of tranexamic acid (TXA) on the reduction of post-operative drained blood volume, pain intensity, and incidence of hemarthrosis after ACL reconstruction., Methods: This prospective research included 124 patients undergoing ACL reconstruction surgery, randomly divided into two groups. The TXA group received IA TXA, whereas an equal amount of placebo was administered using the same route in the control group., Results: The research has shown that IA injection of TXA effectively reduces post-operative blood loss (TXA group 71.29 ± 40.76 vs. control group 154.35 ± 81.45), reducing the intensity of post-operative pain (p < 0.001) and the incidence of hemarthrosis., Conclusion: The application of TXA significantly reduced post-operative bleeding and pain intensity, which accelerated the post-operative period., (Copyright: © 2024 Permanyer.)

Polycystic ovary syndrome (PCOS) is a complex disorder characterized by endocrine and metabolic abnormalities such as obesity and insulin resistance. PCOS is also associated with psychiatric disorders and cognitive impairment. The animal model of PCOS was induced by treating rats with 5α-dihydrotestosterone (5α-DHT) and additionally modified to induce adiposity by litter size reduction (LSR). Spatial learning and memory were assessed using the Barnes Maze test, and striatal markers of synaptic plasticity were analyzed. Striatal insulin signaling was estimated by the levels of insulin receptor substrate 1 (IRS1), its inhibitory phosphorylation at Ser307, and glycogen synthase kinase-3α/β (GSK3α/β) activity. Both LSR and DHT treatment significantly decreased striatal protein levels of IRS1, followed by increased GSK3α/β activity in small litters. Results of the behavioral study showed that LSR had a negative effect on learning rate and memory retention, whereas DHT treatment did not induce impairment in memory formation. While protein levels of synaptophysin, GAP43, and postsynaptic density protein 95 (PSD-95) were not altered by the treatments, DHT treatment induced an increase in phosphorylation of PSD-95 at Ser295 in both normal and small litters. This study revealed that LSR and DHT treatment suppressed insulin signaling by downregulating IRS1 in the striatum. However, DHT treatment did not have an adverse effect on learning and memory, probably due to compensatory elevation in pPSD-95-Ser295, which had a positive effect on synaptic strength. This implies that hyperandrogenemia in this setting does not represent a threat to spatial learning and memory, opposite to the effect of overnutrition-related adiposity., Competing Interests: Declaration of competing interest None., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Aims and Objectives: To evaluate demographic and occupational factors of professional life on life satisfaction among nurses in Serbia and Poland., Background: Compassion satisfaction and compassion fatigue, as aspects of a nurse's professional life, may affect their satisfaction with life in general. However, most studies on this topic have been conducted in Asian countries, and because there are differences in health care system performance across countries amid a global shortage of nurses, it is necessary to investigate professional quality of life in the European nursing context., Methods: This multicenter, comparative, and correlational study was conducted on a sample of 669 hospital nurses in Serbia and Poland. The survey data were collected between November 2019 and February 2020 using the Demographic and Occupational Questionnaire (DOQ), Professional Quality-of-Life Scale (ProQOL-5), and Satisfaction with Life Scale (SWLS). STROBE Statement guidelines for cross-sectional studies were followed., Results: Most nurses from both countries reported having moderate compassion satisfaction, moderate risk for compassion fatigue, and an average life satisfaction. The predictive factors of life satisfaction that most stand out include country of origin, financial situation, education, job satisfaction, and compassion fatigue., Discussion and Conclusion: Quality of professional life is significantly affected by shift duration, age, work experience, and job satisfaction. Negative aspects of professional life are significant predictors of life satisfaction., Implications for Nursing Policy: The findings of this study indicate a need for implementing flexible shift patterns and enhancing interventions that will improve job satisfaction and compassion satisfaction while reducing the risk of compassion fatigue., (© 2022 International Council of Nurses.)

Polycystic ovary syndrome (PCOS) is a common endocrinopathy in women of reproductive age, often associated with obesity and insulin resistance. Childhood obesity is an important predisposing factor for the development of PCOS later in life. Being particularly interested in the interplay between prepubertal obesity and hyperandrogenemia, we investigated the effects of early postnatal overfeeding, accomplished by reducing litter size during the period of suckling, on energy sensing and insulin signaling pathways in the gastrocnemius muscle of a rat model of PCOS-induced by 5α-dihydrotestosterone (DHT). The combination of overfeeding and DHT treatment caused hyperinsulinemia and decreased systemic insulin sensitivity. Early postnatal overfeeding induced defects at critical nodes of the insulin signaling pathway in skeletal muscle, which was associated with reduced glucose uptake in the presence of hyperandrogenemia. In this setting, under a combination of overfeeding and DHT treatment, skeletal muscle switched to mitochondrial β-oxidation of fatty acids, resulting in oxidative stress and inflammation that stimulated AMP-activated protein kinase (AMPK) activity and its downstream targets involved in mitochondrial biogenesis and antioxidant protection. Overall, a combination of overfeeding and hyperandrogenemia resulted in a prooxidative and insulin-resistant state in skeletal muscle. This was accompanied by the activation of AMPK, which could represent a potential therapeutic target in insulin-resistant PCOS patients.

Wastewater-based epidemiology (WBE) surveillance of COVID-19 and other future outbreaks is a challenge for developing countries as most households are not connected to a sewerage system. In December 2019, SARS-CoV-2 RNA was detected in the Danube River at a site severely affected by wastewaters from Belgrade. Rivers are much more complex systems than wastewater systems, and efforts are needed to address all the factors influencing the adoption of WBE as an alternative to targeting raw wastewater. Our objective was to provide a more detailed insight into the potential of SARS-CoV-2 surveillance in Serbian surface waters for epidemiological purposes. Water samples were collected at 12 sites along the Sava and Danube rivers in Belgrade during the fourth COVID-19 wave in Serbia that started in late February 2021. RNA was concentrated using Amicon Ultra-15 centrifugal filters and quantified using RT-qPCR with primer sets targeting nucleocapsid (N1 and N2) and envelope (E) protein genes. Microbiological (faecal indicator bacteria and human and animal genetic faecal source tracking markers), epidemiological, physicochemical and hydromorphological parameters were analysed in parallel. From 44 samples, SARS-CoV-2 RNA was detected in 31, but only at 4 concentrations above the level of quantification (ranging from 8.47 × 10 3 to 2.07 × 10 4 gc/L). The results indicated that surveillance of SARS-CoV-2 RNA in surface waters as ultimate recipients could be used as an epidemiological early-warning tool in countries lacking wastewater treatment and proper sewerage infrastructure. The performance of the applied approach, including advanced sampling site characterization to trace and identify sites with significant raw sewage influence from human populations, could be further improved by adaptation of the methodology for processing higher volumes of samples and enrichment factors, which should provide the quantitative instead of qualitative data needed for WBE., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest., (Copyright © 2022 Elsevier B.V. All rights reserved.)