Cerebrospinal fluid mtDNA concentrations are increased in multiple sclerosis and were normalized after intervention with autologous hematopoietic stem cell transplantation.

BACKGROUND: Mitochondrial DNA (mtDNA) is a pro-inflammatory damage-associated molecular pattern molecule and could be an early indicator for inflammation and disease activity in MS. Autologous hematopoietic stem cell transplantation (aHSCT) is a potent treatment for MS, but its impact on mtDNA levels in cerebrospinal fluid (CSF) remains unexplored. OBJECTIVES: To verify elevated CSF mtDNA concentrations in MS patients and assess the impact of aHSCT on mtDNA concentrations. METHODS: Multiplex droplet digital PCR (ddPCR) was used to quantify mtDNA and nuclear DNA in 182 CSF samples. These samples were collected from 48 MS patients, both pre- and post-aHSCT, over annual follow-ups, and from 32 healthy controls. RESULTS: CSF ccf-mtDNA levels were higher in patients with MS, correlated to multiple clinical and analytical factors and were normalized after intervention with aHSCT. Differences before aHSCT were observed with regard to MRI-lesions, prior treatment and number of relapses in the last year prior to aHSCT. CONCLUSION: Our findings demonstrate elevated CSF mtDNA levels in MS patients, which correlate with disease activity and normalize following aHSCT. These results position mtDNA as a potential biomarker for monitoring inflammatory activity and response to treatment in MS.

Utility of salivary mycophenolic acid concentration monitoring: Modeling and Monte Carlo validation approach.

The results of the previous studies demonstrated an association between mycophenolic acid (MPA) exposure, serum albumin level (ALB), and adverse effects in kidney transplant patients. The aim was the identification of mathematical correlation and association between both, total and unbound MPA concentration in relation to ALB, body mass (BM), age and estimated glomerular filtration rate (eGFR) in stable kidney transplant recipients. Furthermore, investigation was conducted with the aim to clarify the role of salivary concentration (CSAL ) of MPA in adverse effect profile. In order to analyze the association between total and salivary concentration of MPA in relation to ALB, BM, age and eGFR, a least squares method for determining the correlation between these parameters was performed. In addition, derived mathematical model based on experimental data can also be performed and simulated through the Monte Carlo (MC) approach. Adverse effects were grouped according to the nature of symptoms and scored by a previously published validated system. Numerically calculated values of CSAL from the models [CSAL  = f(ALB, BM, age, eGFR, CP ) = a00 + a10 *(ALB, BM, age, eGFR) + a01 *CP ] were then compared with those from validation set of patients, where the best fitting model was for ALB [CSAL  = 54.96-1.64*ALB +13.4*CP ]. Adverse effects estimation showed the difference in esthetic score, positively correlated with CSAL in the lower ALB group (145.41 ± 219.02 vs. 354.08 ± 262.19; with statistical significance p = .014) and almost significant for gastrointestinal score (167.69 ± 174.79 vs. 347.55 ± 320.95; p = .247). The study showed that CSAL MPA may contribute to management of adverse effects, but these findings require confirmation of clinical utility.

Factors Associated With Serological Response to SARS-CoV-2 Vaccination in Patients With Multiple Sclerosis Treated With Rituximab.

Importance: B-cell-depleting monoclonal antibodies are widely used for treatment of multiple sclerosis but are associated with an impaired response to vaccines. Objective: To identify factors associated with a favorable vaccine response to tozinameran. Design, Setting, and Participants: This prospective cohort study was conducted in a specialized multiple sclerosis clinic at a university hospital from January 21 to December 1, 2021. Of 75 patients evaluated for participation who received a diagnosis of multiple sclerosis with planned or ongoing treatment with rituximab, 69 were included in the study, and data from 67 were analyzed. Exposures: Sex, age, number of previous rituximab infusions, accumulated dose of rituximab, previous COVID-19 infection, time since last rituximab treatment, CD19+ B-cell count before vaccination, CD4+ T-cell count, and CD8+ T-cell count were considered potential factors associated with the main outcome. Main Outcomes and Measures: Serological vaccine responses were measured by quantitation of anti-spike immunoglobulin G (IgG) antibodies, anti-receptor-binding domain (RBD) IgG antibodies, and their neutralizing capacities. Cellular responses to spike protein-derived SARS-CoV-2 peptide pools were assessed by counting interferon gamma spot-forming units in a FluoroSpot assay. Results: Among 60 patients with ongoing rituximab treatment (49 women [82%]; mean (SD) age, 43 [10] years), the median (range) disease duration was 9 (1-29) years, and the median (range) dose of rituximab was 2750 (500-10 000) mg during a median (range) time of 2.8 (0.5-8.3) years. The median (range) follow-up from the first vaccination dose was 7.3 (4.3-10.0) months. Vaccine responses were determined before vaccination with tozinameran and 6 weeks after vaccination. By using established cutoff values for anti-spike IgG (264 binding antibody units/mL) and anti-RBD IgG (506 binding antibody units/mL), the proportion of patients with a positive response increased with the number of B cells, which was the only factor associated with these outcomes. A cutoff for the B-cell count of at least 40/µL was associated with an optimal serological response. At this cutoff, 26 of 29 patients (90%) had positive test results for anti-spike IgG and 21 of 29 patients (72%) for anti-RBD IgG, and 27 of 29 patients (93%) developed antibodies with greater than 90% inhibition of angiotensin-converting enzyme 2. No factor associated with the cellular response was identified. Depending on the peptide pool, 21 of 25 patients (84%) to 22 of 25 patients (88%) developed a T-cell response with interferon gamma production at the B-cell count cutoff of at least 40/µL. Conclusions and Relevance: This cohort study found that for an optimal vaccine response from tozinameran, rituximab-treated patients with multiple sclerosis may be vaccinated as soon as possible, with rituximab treatment delayed until B-cell counts have reached at least 40/µL. An additional vaccination with tozinameran should be considered at that point.

Spasmolytic, Gastroprotective and Antioxidant Activities of Dry Methanol Extract of Ferula heuffelii Underground Parts.

Dry MeOH extract of Ferula heuffelii (Apiaceae) underground parts was tested for spasmolytic, gastroprotective and antioxidant activities. HPLC analysis revealed that chlorogenic acid (CGA; 34.6 mg/g) was its main constituent. Extract in vitro exhibited notable total antioxidant activity (FRAP value=1.0 µmol Fe2+ /mg), and scavenging of DPPH (SC50 =62.5 µg/ml) and • OH radicals (49.5 % at 20 µg/ml in 2-deoxyribose assay). In vitro on isolated rat ileum, extract exhibited significant spasmolytic activity, i. e., it showed 124.6 % of maximal atropine effect on spontaneous contractions (at 100 µg/ml), and reduced spasmogenic effect of KCl (80 mm) to 44.4 % (at 60 µg/ml) and of highest applied concentration of ACh to 26.3 % (at 120 µg/ml). In parallel experiments, spasmolytic effect of CGA was also demonstrated. In acute EtOH-induced gastric ulceration model in rats, extract (100 mg/kg p.o.) showed significant gastroprotective effect (gastric damage score 0.50), similar to ranitidine (20 mg/kg p.o.). Obtained results showed that tested F. heuffelii polar extract represents new herbal preparation with potential use against some gastrointestinal complaints.

The presence of deer ked (Lipoptena cervi, Linnaeus, 1758) in Balkan chamois from the National Park Sutjeska, Bosnia and Herzegovina.

This case study describes the presence of a deer ked (Lipoptena cervi, Linnaeus, 1758) in the Balkan chamois (Rupicapra rupicapra balcanica) from the area of the National Park "Sutjeska", Bosnia and Herzegovina (B&H). This is the first finding of this ectoparasite in this animal species in B&H, which indicates that deer ked has significant adaptive abilities to different animal hosts.

Effect of the Interrelation between CYP3A5 Genotype, Concentration/Dose Ratio and Intrapatient Variability of Tacrolimus on Kidney Graft Function: Monte Carlo Simulation Approach.

Background: Tacrolimus (Tac) is characterized by large between- and within-patient (IPV) variability in pharmacokinetics and exposure. Aim: This study aimed to assess and validate the effect of Tac IPV and trough concentration-to-dose ratio (C0/D) over 6-12 months on reduced estimated glomerular filtration rate (eGFR) values in the late period after kidney transplantation (Tx), applying Monte Carlo (MC) simulation. Methods: The previously published linear regression was the basis for MC simulation, performed to determine how variations in significant predictors affect the distribution of eGFR from 13 to 36 months post-transplantation. The input C0/D values were derived from CYP3A5 genotype subgroups. Results: Patients characterized by high Tac IPV and low mean C0/D over 6-12 months could have been at greater risk of lower eGFR values in a three-year period following Tx compared to the other patient groups. This effect was more pronounced in patients with a lower eGFR at the 6th month and a history of acute rejection. The proven contribution of CYP3A5 expresser genotype to low C0/D values may suggest its indirect effect on long-term graft function. Conclusion: The findings indicate that simultaneous assessment of Tac IPV, C0/D, and CYP3A5 genotype may identify patients at risk of deterioration of graft function in the long-term post-transplantation period.

Prevalence, Intensity and Risks Involved in Helminth Infections in Domestic Mountain Pony and Balkan Donkey in Nature Park Stara Planina, Serbia.

BACKGROUND: This study aimed to investigate the prevalence, intensity and risk factors (age and gender) promoting the intestinal helminthic infections of the Domestic mountain ponies and Balkan donkeys in Serbia. METHODS: Prevalence, intensity and risk factors influencing helminth infection in horses (n=39) and donkeys (n=18) at the site of Nature Park Stara Planina, were studied from May to Sep 2015. The presence of one or several gastrointestinal helminth species was confirmed by faecal flotation in 97.43% of horses and 88.88% of donkeys included in the monitoring. The identified helminth species were Trichostrongylus axei, Strongylus edentatus, S. vulgaris, Parascaris equorum, Dictyocaulus arnfieldi and Anoplocephala magna in 84.61%, 46.15%, 5.13%, 58.97%, 94.87% and 38.46% of horses, respectively. The significant association of infection with P. equorum and sex of horses was established. (χ2=13.33, P<0.01). RESULTS: The prevalence of parasitic helminths identified in donkeys was the following: D. arnfieldi, T. axei, Pa. equorum, S. vulgaris, A. margna and Strongyloides westeri in 55.55%, 38.89%, 27.78%, 22.22% and 22.22% of donkeys, respectively. Moreover, the mean faecal egg count was higher in donkeys (369.9 EPG - egg per gram) than in horses (250.2 EPG). The association between the age and the mean EPG was significant (P<0.05) in both equine species. CONCLUSION: The results of the investigation provided basic information that can be helpful for planning strategic control of nematode infection in equine population in Nature Reserves in Serbia.

Immunohistochemical expression of cyclin-dependent kinase inhibitors p16 and p57 in rhabdomyosarcoma.

Rhabdomyosarcoma (RMS) is a highly malignant cancer and is the most common soft tissue sarcoma in children and adolescents, but it is rare in adults (<1% of all adult malignancies). Altered expression and molecular abnormalities of cell-cycle-regulatory proteins are one of the most prominent features in RMS. Therefore, we evaluated the expression of cyclin-dependent kinase inhibitors p57 and p16, as well as p16 methylation status, along with clinicopathological characteristics and overall survival (OS) in RMS patients. This analysis was conducted on 23 pediatric and 44 adult patients. There was a male predominance in both groups and extremities were the most frequent tumor site. In adults, alveolar and pleomorphic types were almost equally represented. The majority of pediatric tumors were low grade, whereas, in adults, only one patient had a low-grade tumor. Seven pediatric (30.43%) and eight adult (18.18%) patients had a low p16 expression. The analysis of methylation status of the p16 promoter showed the presence of methylated allele only in one sample with pleomorphic histology. Six (26.1%) pediatric and 15 (34.1%) adult patients had low p57 expression, while in 17 (73.9%) pediatric and 29 (65.9%) adult patients it was assessed as high. Ninetyone percent of the pediatric patients and 32.6% of adults were alive at the end of the observational period. In adults, significant associations were found between OS and age (P = 0.020), gender (P = 0.027), tumor size (P < 0.001), lymph node status (P < 0.001), presence of metastases (P = 0.015), and p57 expression (P = 0.039). Stratification by histological type showed the correlation of low p57 expression (P = 0.030) and worse OS of patients with alveolar RMS. Univariate analysis identified age > 50 yrs. (HR 2.447), tumors > 5 cm (HR 21.31), involvement of regional lymph nodes (HR 3.96), the presence of metastases (HR 2.53), and low p57 expression (HR 2.11) as predictors of lower OS. Tumor size, regional lymph nodes involvement, and metastases were the independent predictors after multivariate analysis, while p57 did not predict OS in an independent way. In summary, although p57 was not confirmed to be an independent predictor of OS, our results indicate that its low expression may be the marker of aggressive phenotype and poor prognosis in adult RMS patients. Also, our findings suggest that epigenetic inactivation of p16 is not important in the pathogenesis of rhabdomyosarcoma.

Effects of mood stabilizer lithium on noradrenergic turnover in the prefrontal cortex of chronically stressed rats.

OBJECTIVE: Data about the dynamics of noradrenaline (NA) transmission, storage and degradation may be very important for understanding the reduction of functional deficiency of NA and maintaining the stability of NA levels in animals with depressive-like behavior treated with lithium. This study aimed to investigate the effects of mood stabilizer lithium on concentrations of NA in the prefrontal cortex (PFC), as well as behavior rats exposed to chronic restraint stress (CRS). In addition, this study examined the effects of lithium on protein levels of noradrenaline transporter (NET), vesicular monoamine transporter 2 (VMAT2) and catechol-O-methyltransferase (COMT), as well as the enzyme activity of monoamine oxidase A (MOA) in the PFC of chronically stressed rats. METHODS: The investigated parameters were quantified by Western blot analysis, CAT Research ELISA kits, and an assay of enzyme activity. Also, the forced swim test (FST) was used to assess the behavior of animals. RESULTS: We found that lithium treatment decreased high protein levels of NET and VMAT2, as well as the enzyme activity of MOA in chronically stressed rats to the levels found in unstressed animals. In addition, lithium treatment decreased the concentration of NA (24%) and immobility in animals exposed to CRS. CONCLUSION: Our data confirm that lithium-induced modulation of prefrontal noradrenergic turnover and stabilized the behavior of chronically stressed rats.

Side-by-side evaluation of virtual non-contrast and post-contrast images improves detection of clinically significant urolithiasis on single-phase split bolus dual-energy CT urography.

OBJECTIVES: Studies show insufficient sensitivity of virtual non-contrast (VNC) reconstructions for stone detection in dual-energy CT urography (DE-CTU). The aim of this study was to investigate if side-by-side-evaluation of both VNC and post-contrast images could increase the sensitivity of single-phase split bolus DE-CTU. METHODS: Consecutive patients with haematuria who underwent split bolus DE-CTU on the same dual-source DE-CT scanner were retrospectively enrolled in the study. Intravenous furosemide and oral hydration were employed. Two readers, independently and then jointly in two separate sessions, recorded the location and the longest axial stone diameter on three randomised sets of images: separate VNC and post-contrast images, and side-by-side-reconstructions. True non-contrast (TNC) images served as the standard of reference. RESULTS: A total of 83 urinary stones were detected on TNC images. Independent reader side-by-side-evaluation of VNC and post-contrast images yielded higher stone detection sensitivity (76 and 84%, respectively) compared to evaluation of only VNC (71 and 81%, respectively) or post-contrast images (64 and 80%, respectively). The sensitivity of joint reader evaluation of side-by-side-images reached almost 86% and was not significantly different from TNC images (p = 0.77). All stones larger than 3 mm were correctly detected by side-by-side-evaluation. Dose reduction of 55% could be achieved by omitting TNC scans. CONCLUSION: Side-by-side-VNC and post-contrast image evaluation enable detection of clinically significant urolithiasis on single-phase split bolus DE-CTU with significant dose reduction. ADVANCES IN KNOWLEDGE: This study shows that single-phase DE-CTU is feasible if VNC imaging is simultaneously utilised with post-contrast images.

Canine intestinal parasites as a potential source of soil contamination in the public areas of Krusevac, Serbia.

INTRODUCTION: Environmental contamination by infected canine faeces presents an indirect source of contamination in people. In this research the presence of parasitic elements in canine faeces found in the public areas of Krusevac was examined. METHODOLOGY: During May and October 2018, 282 samples were collected from different public areas (ten kindergartens, six public squares and four parks). The examination of faeces was performed by means of direct smear and flotation. For statistical testing of the difference between the frequencies of attributes χ2 test was applied. RESULTS: Out of 282 samples, 221 (78.4%) were positive, of which 17/20 (85.0%) from kindergartens, 125/160 (78.1%) from parks and 79/102 (77.5%) from public squares. The presence of Ancylostomatidae spp. was discovered in 113 (40.1%) samples, Toxocara canis in 82 (29.1%), Dipylidium caninum in 76 (27.0%), Giardia intestinalis in 45 (16.0%), Taenia spp. in 44 (15.6%), Amoeba spp. in 32 (11.3%), Trichuris vulpis in 19 (6.7%), Toxascaris leonina in 18 (6.4%), Strongyloides stercoralis in 17 (6.0%), Isospora spp. in 7 (2.5%) and Cryptosporidium spp. in 3 (1.1%). In May there were 115/141 (81.6%) positive samples, whereas in October there were 106/141 (72.5%) positive ones. No positive sample with three or four parasites was found in October. CONCLUSIONS: This study shows that canine faeces from public areas in Krusevac can be the cause of a significant environmental contamination by eggs of canine intestinal parasites as well as of human infections.

Assessment of pharmacokinetic mycophenolic acid clearance models using Monte Carlo numerical analysis.

Previously, we performed population pharmacokinetic analysis and indicated age, mycophenolate mofetil (MMF)/mycophenolic acid (MPA) daily dose, and presence of nifedipine in patient therapy as significant predictors of MPA apparent clearance (CL/F) variability. This study aimed to determine the reliability of previously published population pharmacokinetic models derived from similar studies. Furthermore, this study investigated correspondence between chosen population models from the literature.By means of the Monte Carlo simulation method, pharmacokinetic models from different studies are simulated and analysed in the range of standard deviations of measured system parameters as well as the range of observed model parameters taken from the comparison studies.The 1000 numerical simulations were performed for every analysed model in order to calculate the most possible MPA CL/F values according to the expected values from the performed experiment. Fitting our results with other models showed how the presence of nifedipine makes difference in MPA CL/F values.By testing the data from selected studies into our model, a similar range of expected CL/F values was obtained, which may confirm the validity of our model. The results of our population pharmacokinetic study are partially applicable in models by other researchers.

The effect of antioxidant status on overall survival in renal cell carcinoma.

INTRODUCTION: The oxidative stress contributes to all three phases of carcinogenesis and represents a concomitant condition in renal cell carcinoma (RCC). RCC is the most common type of neoplasm of the kidney, and despite numerous studies the set of predictive and prognostic markers of survival are still unknown. The aim of our study was to examine the relation between antioxidant (AO) status and overall survival (OS) in RCC patients. MATERIAL AND METHODS: Our study included 95 patients with RCC, who underwent radical nephrectomy. We analysed the prognostic role of antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase, glutathione reductase, glutathione, and malondialdehyde) and other clinicopathological factors (size, grade, stage, and histological subtype) on the OS of RCC patients. RESULTS: The 5-year OS was 54.6%. The survival analysis related to AO parameters showed no significant difference in survival of RCC patients. The concentration of malondialdehyde, an indicator of lipid peroxidation, also had no significant effect on the survival rate of RCC patients. Univariate and multivariate analysis confirmed the significance of clinicopathological parameters (size, p < 0.001; Fuhrman grade, p = 0.001, and stage, p < 0.001) for patients' survival. CONCLUSIONS: In our cohort of patients, different antioxidant parameters were not found to be predictors for OS of patients with RCC, who underwent radical nephrectomy.

Influence of breed on selected quality parameters of fresh goat meat.

The potential of goats to produce a high-quality meat is mainly reflected in their healthy fats, low calorie intramuscular fats, saturated fats, and, especially, their high ratios of unsaturated (UFA) and saturated (SFA) fatty acids, as well as hypocholesterolemic and hypercholesterolemic fatty acids. The aim of this study was to collect and compare meat quality parameters for domestic Balkan, Alpine and Saanen goats of the same age. Samples for all tests were taken from musculus gluteus superficialis. Chemical composition, pH value, fatty acid composition, content of volatile compounds, color and overall sensory quality (appearance, texture and smell) were determined. In chemical composition, moisture, fat, protein and ash varied significantly between each of the examined groups as opposed to pH values. Furthermore, among all the examined groups a significant difference was found for fatty acids and volatile compounds. Determined ratio of polyunsaturated fatty acids (PUFAs) to SFAs was 0.089, 0.085 and 0.071 for Balkan, Alpine and Saanen goat meats, respectively. Regarding that ratio, Saanen goat meat had the most favorable characteristics. Saanen goat meat showed the highest nutritional value. On the other hand, Balkan goat meat had the lowest intramuscular fat content. Measurements of the meat color from all three groups, as well as overall acceptability, showed significant differences between breeds. Obtained results point to the impact of breed on chemical composition and fatty acid profile of goat meat.

Beneficial Role of ROS in Cell Survival: Moderate Increases in H2O2 Production Induced by Hepatocyte Isolation Mediate Stress Adaptation and Enhanced Survival.

High levels of reactive oxygen species (ROS) can lead to impairment of cell structure, biomolecules' loss of function and cell death and are associated with liver diseases. Cells that survive increased ROS often undergo malignant transformation. Many cancer cells tolerate high levels of ROS. Here we report a transiently increased production of H2O2 and concomitant upregulation of antioxidative enzymes triggered by hepatocyte isolation; the H2O2 levels revert in about two days in culture. Three-day survival rate of the isolated cells in the presence of 2.5-fold increase of H2O2 is almost 80%. Apoptosis activation through the mitochondrial pathway is meanwhile reduced by inhibition of caspase-9 triggering. This reduction depends on the amount of H2O2 production, as decreased production of H2O2 in the presence of an antioxidant results in increased apoptosis triggering. These stress adaptations do not influence urea production, which is unchanged throughout the normal and stress adapted phases. We conclude that hepatocytes' stress adaptation is mediated by increased ROS production. In this case, high ROS improve cell survival.

Evidence of West Nile Virus (WNV) Circulation in Wild Birds and WNV RNA Negativity in Mosquitoes of the Danube Delta Biosphere Reserve, Romania, 2016.

West Nile virus (WNV) is a zoonotic flavivirus whose transmission cycle in nature includes wild birds as amplifying hosts and ornithophilic mosquito vectors. Bridge vectors can transmit WNV to mammal species potentially causing West Nile Fever. Wild bird migration is a mode of WNV introduction into new areas. The Danube Delta Biosphere Reserve (DDBR) is a major stopover of wild birds migrating between Europe and Africa. The aim of this study was to investigate the presence of WNV in the DDBR during the 2016 transmission season in wild birds and mosquitoes. Blood from 68 wild birds (nine different species) trapped at four different locations was analyzed by competitive ELISA and Virus Neutralization Test (VNT), revealing positive results in 8/68 (11.8%) of the wild birds by ELISA of which six samples (three from juvenile birds) were confirmed seropositive by VNT. Mosquitoes (n = 6523, 5 genera) were trapped with CDC Mini Light traps at two locations and in one location resting mosquitoes were caught. The presence of WNV RNA was tested in 134 pools by reverse transcription quantitative PCR (RT-qPCR). None of the pools was positive for WNV-specific RNA. Based on the obtained results, WNV was circulating in the DDBR during 2016.

Serum Concentrations and Pharmacokinetics of Tranexamic Acid after Two Means of Topical Administration in Massive Weight Loss Skin-Reducing Surgery.

BACKGROUND: Topical administration of tranexamic acid to reduce bleeding is receiving increasing attention, as it is inexpensive, simple, and possibly beneficial in most surgery. Concerns regarding potential systemic adverse effects such as thromboembolic events and seizures may prevent general use of tranexamic acid. Although serum concentrations after topical application are assumed to be low, proper pharmacokinetic studies of tranexamic acid after topical application are lacking. METHODS: The authors have investigated systemic absorption of tranexamic acid after two means of topical administration in patients undergoing abdominoplasty after massive weight loss: a bolus of 200 ml of 5 mg/ml into the wound cavity versus moistening the wound surface with 20 ml of 25 mg/ml. Twelve patients were recruited in each group. Serum concentrations achieved were compared with those after administration of 1 g as an intravenous bolus to arthroplasty patients. Serial blood samples for tranexamic acid analysis were obtained for up to 24 hours. RESULTS: After intravenous administration, the peak serum concentration was 66.1 ± 13.0 µg/ml after 6 ± 2 minutes. Peak serum concentration after topical moistening was 5.2 ± 2.6 µg/ml after 80 ± 33 minutes, and in the topical bolus group, it was 4.9 ± 1.8 µg/ml after 359 ± 70 minutes. Topical moistening resulted in homogenous and predictable absorption across the individuals included, whereas topical bolus administration caused variable and unpredictable serum concentrations. CONCLUSION: Topical administration of tranexamic acid in patients undergoing abdominoplasty results in low serum concentrations, which are highly unlikely to cause systemic effects.

Alaria alata mesocercariae in domestic pigs and wild boars in South Banat, northern Serbia.

Alaria alata (Diplostomidae, Trematoda), a potentially zoonotic pathogen, is the only Alaria species in Europe. In recent years, increasing reports of mesocercariae of A. alata in wild boars have been recorded in European countries; however there have been no described cases in domestic pigs over the past decades. Out of 272 diaphragm samples from wild boars (Sus scrofa) and domestic pigs (Sus scrofa), from Serbia, included in the present investigations, mesocercariae of A. alata were found in six (3%) samples from wild boars and in two (2.77%) samples from domestic pigs. Due to the lack of sensitivity of artificial digestion with a magnetic stirrer which was applied in the study, the true prevalence is estimated to be considerably higher in the area of investigation. Confirming the infection in domestic pigs and a wide distribution in wild boars in the area of investigation, the present paper should influence the growing awareness of alariosis as a potential human disease. The meat from free-range domestic pigs and game meat as a potential source of alariosis should always be adequately thermally treated before consumption.

Cardiopulmonary parasitic nematodes of the red fox (Vulpes vulpes) in Serbia.

Among the wild canids, the red fox (Vulpes vulpes) is recognised as an important reservoir species for a range of parasites, including cardiopulmonary nematodes of public health and veterinary importance. As cross-host transmission between the red fox and domestic carnivores can play an important role in the epizootiology of cardiopulmonary parasitic diseases, the aim of the present investigations was to obtain data on the geographical distribution of cardiopulmonary nematodes of the red fox. The material for examination consisted of 83 foxes which were legally hunted at different locations during a three-month period from December 2017 to February 2018. The presence of four emerging species in Europe (Angiostrongylus vasorum, Crenosoma vulpis, Dirofilaria immitis and Eucoleus aerophilus) was revealed in red foxes of Serbia. Crenosoma vulpis and E. aerophilus were detected in foxes in both the plain and the mountainous areas across the country. Dirofilaria immitis is distributed in red foxes near alluvial rivers in Vojvodina province (northern Serbia). Angiostrongylosis caused by A. vasorum was demonstrated to exist in two enzootic foci with a high percentage of infected foxes in a plain area of northern Serbia. To the best of our knowledge, C. vulpis and A. vasorum were discovered for the first time in red foxes in central Serbia. The results provide strong evidence for veterinarians to take into consideration the parasitic nematodes discovered in red foxes in the differential diagnosis of diseases of companion animals. In the context of the 'One Health' approach the results related to the distribution of the zoonotic species E. aerophilus and D. immitis can be useful for medical epidemiology.

Cyclin D1 and p57 expression in relation to clinicopathological characteristics and overall survival in patients with renal cell carcinoma.

PURPOSE: There is a need for identifying molecular prognostic biomarkers to better predict clinical outcomes in patients with renal cell carcinoma (RCC). This study investigated the pattern of cyclin D1 and p57 expression in RCC patients and evaluated their relation with clinicopathological characteristics and overall survival (OS). METHODS: Immunohistochemistry was applied to paraffin-embedded tissue sections of 74 RCC patients. Two cut-off groups were defined by the fraction of positive cells as follows: ≤10% and >10% positive cells for cyclin D1, and ≤5% and >5% positive cells for p57. RESULTS: Cyclin D1 expression in >10% of positive cells was observed mostly in the clear cell RCC, while p57 expression in ≤5% of positive cells was found in 86% of chromophobe RCC specimens. The higher expression of cyclin D1 and lower expression of p57 were more frequent in grade I-II tumors. OS was associated with unfavorable clinicopathological characteristics. However, cyclin D1/p57 expression did not influence the survival rates. CONCLUSION: Although cyclin D1 and p57 expression did not affect survival rates in RCC patients, proper validation and establishment of the qualitative cut-off point are needed for these tumors.