The prevalence of obesity worldwide has rapidly increased. Numerous studies showed a beneficial effect of probiotics in obese individuals, and changes in hematological parameters are observed in obesity. Therefore, the aim of this study was to investigate the effect of a novel probiotic approach on the red blood cells (RBCs) and platelets. Twenty-five obese women participated in a randomized, placebo-controlled study and were divided into the experimental group (one capsule daily containing Lactiplantibacillus plantarum 299v (DSM9843), Saccharomyces cerevisiae var. boulardii, and 40 mg octacosanol; n = 13) and the placebo group (n = 12). Blood samples were collected for light microscopic examination, morphometric analysis, and an automated hematology analyzer. A possible relationship between hematological parameters and body mass index (BMI), a common indicator of obesity, was investigated using Spearman correlation. The plasma concentration of soluble P-selectin and fibrinogen were determined using an ELISA assay. All measurements were performed before (T0) and after 12 weeks of supplementation (T1). The three-month supplementation of probiotics improved hemoglobin levels, chromic status, and red blood cell morphology. The mean platelet volume (MPV), a measure of platelet size, was restored to normal levels, platelet morphology was improved, and the number of activated platelets was significantly reduced (p < 0.05). A strong negative correlation (r = -0.5904, p < 0.05) was found between BMI and platelet distribution width (PDW), a measure of variation in platelet size and shape. The results show that the probiotic approach improves morphology and normalizes the values of disturbed hematological parameters of RBCs and platelets in obese women.

Clear-cell renal cell carcinoma (ccRCC) is associated with the mutated von Hippel-Lindau (VHL) gene leading to the activation of hypoxia-inducible factor 1A (HIF1A) and subsequent overexpression of erythropoietin (EPO). We analyzed tumor and healthy tissues from 43 ccRCC patients after radical nephrectomy and cultured 786-O (biallelic VHL inactivation) and Caki-1 (wild-type VHL) cells in normal (21% O2) and low oxygen (3% O2) with 10% and 2% fetal bovine serum (FBS). DNA sequencing, including Sanger sequencing, MLPA and LOH, revealed 27 somatic mutations of VHL in ccRCC. HIF1A protein showed decreased or no expression in tumors compared to healthy tissue, independent of VHL alteration. The 786-O cells showed increased HIF1A protein expression after 48 h under low oxygen and 10% FBS. EPO and erythropoietin receptor (EPOR) were significantly decreased in ccRCC without HIF1A expression. EPO mRNA increased in the 786-O cells at 3% O2 after 48 h, while the Caki-1 cells had low or no EPO expression. Hypoxia increased EPOR mRNA in the Caki-1 cells at 10% FBS, but decreased in the 786-O cells at 2% FBS after 48 h. JAK2/STAT5A activity was increased only in HIF1A-positive tumors. These results suggest that EPO/EPOR activation in ccRCC is mainly driven by low oxygen, not VHL regulation of hypoxia-related responses.

Background: The global burden of inflammatory bowel diseases (IBDs), including ulcerative colitis and Crohn's disease, is constantly rising. As IBDs significantly reduce patients' quality of life, prevention and efficient treatment of IBDs are of paramount importance. Although the molecular mechanisms underlying IBD pathogenesis are still not completely understood, numerous studies indicate the essential role of oxidative stress in the progression of the diseases. Objective: The aim of this study was to investigate whether prophylactic administration of recombinant banana lectin (rBanLec) could positively affect antioxidative mechanisms in the colon and thus prevent or alleviate the severity of experimental colitis induced in C57BL/6 mice. Methods: The prophylactic potential of rBanLec, a mannose-binding lectin with immunomodulatory properties, was investigated in a model of 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis in C57BL/6 mice. Mice received rBanLec at various doses (0.1, 1 and 10 μg/mL) before the induction of colitis. The severity of the disease was assessed by weight loss and reduction in colon length, and correlated with histopathological findings, cytokine milieu, and oxidative stress markers in the colon. Results: The obtained results revealed that pretreatment with a low dose of rBanLec (0.1 μg/mL) significantly reduced the severity of TNBS-induced colitis, as indicated by reduced weight loss, less severe histopathological damage, and a favorable anti-inflammatory cytokine milieu (increased IL-10 and TGFβ). In addition, rBanLec pretreatment improved the activity of antioxidant enzymes (SOD, CAT, and GST) and reduced markers of oxidative stress such as nitric oxide levels at the peak of the disease. In contrast, higher doses of rBanLec exacerbated inflammatory responses. Conclusions: Our findings indicate that at low doses rBanLec can alleviate the severity of colitis by modulating oxidative stress and promoting anti-inflammatory cytokine responses, positioning rBanLec as a potential candidate for treating IBDs.

Finding and characterizing new bacterial strains, including probiotic strains, is a crucial task in today's world to expand the genetic data pool and identify new genes. In this study, we investigated the gut microbiota of one industrial species, Cyprinus carpio, and identified representatives of various microbial genera, including Citrobacter, Serratia, Bacillus, Enterococcus, and Kocuria. Notably, we discovered two strains of Hafnia with potentially probiotic properties. We conducted next-generation sequencing (NGS) of these strains, described their antibiotic resistance and antibacterial activity, and compared them with other representatives of the Hafnia genus. These strains, characterized by rapid growth, the presence of the ClpB heat shock protein gene, and genes associated with microplastic degradation, provide a promising basis for further research, including studies on their potential application in plastic biodegradation.

Deep proteomic analyses identified, in total, 159 master proteins (with 1% FDR and 2 unique peptides) from 26 protein families in the venom of Vipera ammodytes ammodytes (Vaa). Data are available via ProteomeXchange with the identifier PXD056495. The relative abundance of PLA2s is 11.60% of the crude venom, of which 4.35% are neurotoxic Ammodytoxins (Atxs). The neurotoxicity of the venom of Vaa and the neutralizing effect of the antivenom were tested on the neuromuscular preparation of the diaphragm (NPD) of rats. The activity of PLA2 in the venom of Vaa and its neutralization by the antivenom were determined under in vitro conditions. The Vaa venom leads to a progressive decrease in NPD contractions. We administered pre-incubated venom/antivenom mixtures at various ratios of 1:2, 1:10 and 1:20 (w/w) and observed the effects of these mixtures on NPD contractions. The results show that the mean effective time (ET50) for NPD contractions with the 1:20 mixture is highly significantly different (p < 0.001) from the ET50 for the venom and the ET50 for the 1:2 and 1:10 mixture ratios. We also found a highly significant (p < 0.001) reduction in Na+/K+-ATPase activity in the NPD under the influence of the venom. The reduction in the activity of this enzyme was reversible by the antivenom. Under in vitro conditions, we have achieved the complete neutralization of PLA2 by the antivenom. In conclusion, the antivenom abolished the venom-induced progressive decrease in NPD contractions in a concentration-dependent manner. Antivenom with approximately the same mass proportion almost completely restores Na+/K+-ATPase activity in the NPD and completely neutralizes the PLA2 activity of the venom in vitro.

The progression of monoclonal gammopathies is affected by a range of factors, including the microenvironment surrounding plasma cells. It is recognized that TGF-β1 plays a distinct role in stimulating IgA production. Hence, this study aims to investigate whether individuals with serum IgA monoclonal immunoglobulins (paraproteins) exhibit elevated total TGF-β1 levels compared to those with IgG or IgM paraproteins. To achieve this goal, individuals with a positive laboratory finding of monoclonal gammopathy were segregated according to the paraprotein class as well as according to the type of the light chain. Total TGF-β1 levels were assessed in blood serum samples containing IgG (n = 50), IgA (n = 46), and IgM (n = 31) paraproteins. Elevated level of TGF-β1 was confirmed in sera with IgA paraproteins (median 25.8 ng/mL; interquartile range IQR: 19.0-33.7) compared to those having IgG (median: 18.2 ng/mL; IQR: 14.3-22.1; p < 0.001) or IgM paraproteins (21.5 ng/mL; IQR: 15.0-27.4; p = 0.043). Also, a higher TGF-β1 level was detected in sera with IgMλ than those with IgMκ paraproteins (p = 0.043). This research affirms the role of TGF-β1 in the pathophysiology of IgA monoclonal gammopathies and the potential switch towards the IgA isotype, known for a less favourable prognosis.

Abstract Introduction Integrated sentinel SARI surveillance of influenza, SARS -CoV-2 and RSV has been established in Serbia since 2021-2022 influenza season. Nine sentinel hospitals in 3 cities participated in the SARI surveillance system. The aim of this study is to provide a review of integrated sentinel surveillance of SARI in Serbia in 2023-2024 influenza season, from October 2023 to April 2024. Methods On a weekly basis, both epidemiological and virological data were collected. Surveillance is be conducted year-round. Specimens were tested for influenza, SARS-CoV-2,RSV and other respiratory viruses. Respiratory disease wards, intensive care units (ICUs) and pediatric wards were all represented. Real time polymerase chain reaction (RT-PCR) was used. Results From October 2023 to April 2024, a total of 1571 SARI cases were reported with 100% recorded age. Of these, 35% were 0-4 years old, 16% were 5-14, 6% were 15-29, 14% were 30-64, 18% were 65-79 and 11% were 80+. Among these cases, 1002 (64%) respiratory specimens were collected. The number of positive samples for influenza was 294 (29%). The positivity rate was 10% for SARS-CoV-2 and 19 % for RSV. The highest proportion of laboratory-confirmed influenza cases was 67% in week 04/2023. Then above 50% in week 05/2023 and 06/2023. Type A viruses was predominated, accounting for 97% of all influenza SARI detections. Activity was dominated by A(H3) with 42% positivity rate. Also, collected swabs were tested for rhinoviruses, positivity rate was 26%, for adenoviruses, 12%, for human metapneumovirus, 7% and for parainfluenza 3 viruses, 4%. Conclusions Sustaining and intensified effective integrated surveillance for influenza, SARS-CoV-2,RSV and other respiratory viruses in Serbia is the priority especially in the frame of International Health Regulations. In this way, we improve national capacity for preparedness and emergency response to national and cross-border public health threats. Key messages • Building and maintaining integrated respiratory surveillance that can rapidly integrate the surveillance of a new viruses is crucial. • Establishing well structured surveillance of severe acute respiratory infections, strengthening and expanding respiratory viral surveillance is one the base pillars preparation for future pandemics.

Abstract Issue/problem To date, Serbia has not detected locally acquired Dengue cases. Since a competent Dengue virus vector Aedes albopictus (Ae. albopictus) has been detected since 2017, imported cases have the potential to trigger local transmission. Description of the problem To ensure preparedness and timely response in the event of an epidemic signal in Serbia, we described the imported Dengue cases and analysed the geographical distribution and density of the vector. Probable and confirmed imported cases meeting the 2018 EU case definition were reported via the National Notifiable System between January 2019 and December 2023. We monitored the density and geographical spread of mosquitos, including Ae. albopictus, which were collected during May-October 2022 and 2023. Mosquito genera were determined according to morphological criteria and DNA barcoding. Results During the investigation period, physicians reported three probable and four confirmed imported Dengue cases; the mean age was 28.4 (standard deviation (SD)=11.5) years and 57% were male. Cases had travelled in Southeast Asia (n = 6) and the Caribbean (n = 1). The mean time interval from symptom onset to seeking medical care was 3.6 (SD = 2.1) days and from symptom onset to laboratory confirmation 7.4 (SD = 6.3) days. We collected 1994 Ae. albopictus mosquitoes in 308 sampling sites across 12 Serbian regions. We observed the highest number of mosquitoes in September (38%, p &amp;lt; 0.001) and in the Belgrade region (81%, p &amp;lt; 0.001). In 2022, we detected Ae. albopictus mosquitos in seven regions and in 2023 in nine. Lessons Ae. albopictus mosquitoes were widely distributed in Serbia, with the highest presence during September and in the capital region. The importation of Dengue cases from affected countries underscores the need for enhanced integrated and “real-time” surveillance linked with vector control activities.

This study aimed to investigate the probiogenomic features of artisanal bacteriocin-producing Enterococcus faecium BGPAS1-3 and the use of the improved pMALc5HisEk expression vector for overexpressing class II bacteriocins and the application of purified bacteriocin 31 in a milk model as a preservative against L. monocytogenes. The BGPAS1-3 strain was isolated from traditional fresh soft cheese manufactured in households on a small scale in rural locations surrounding Pale Mountain City in Bosnia and Herzegovina. The whole-genome sequencing approach and bioinformatics analyses revealed that the strain BGPAS1-3 was non-pathogenic to humans. The presence of bacteriocin operons suggested the ability of the isolate to suppress the growth of pathogens. Coding regions for three maturated bacteriocins (bacteriocin 31, bacteriocin 32, and enterocin P) produced by BGPAS1-3 were amplified and expressed in Escherichia coli ER2523 using the pMALc5HisEk system. All three bacteriocins were successfully overexpressed and purified after enterokinase cleavage but showed different antimicrobial activity. Bacteriocin 31 showed significantly stronger antimicrobial activity compared with bacteriocin 32. It was the only one that proved to be suitable for use as a food preservative against L. monocytogenes in a milk model.