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Objective. To investigate the frequency of isolates of various uropathogens in patients of the COVID-19 Hospital "Mišeluk" in comparison with patients from the nonCOVID-19 zone of the Clinical Center of Vojvodina (KCV). Methods. For the purpose of this study, information was collected on urine isolates sent from the COVID-19 hospital and the non-COVID-19 zones of the Clinical Center of Vojvodina in a three-month period, with samples from the non-COVID-19 zone KCV used as controls. Results. Of the 449 patients from the COVID-19 Hospital, the most common causative agents of intrahospital urinary infection (IUI) were members of the genus Enterococcus, (216/449; 48.10%), which in most cases caused IUI in women (127/216; 58.79%). The second most common causative agent of IUT in COVID-19 was Klebsiella pneumoniae (85/449; 18.93%). Of the 913 patients from the non-COVID-19 KCV clinics, the most common cause of IUI was Escherichia coli (249/913; 27.27%), predominantly in women (199/249; 79.91%, (ch2 (1) = 49,042). p

The tick-borne pathogens (TBPs) with adhesive phenotype can use platelets for dissemination and colonization of distant tissues and organs, and it has been shown that they can be found concentrated in the platelet fraction of blood. This study shows the differential presence of TBPs in samples of human platelet fraction (n = 68), whole blood samples (n = 68) and ticks collected (n = 76) from the same individuals, using an unbiased high-throughput pathogen detection microfluidic system. The clinical symptoms were characterized in enrolled patients. In patients with suspected TBP infection, serological assays were conducted to test for the presence of antibodies against specific TBPs. Tick species infesting humans were identified as Ixodes ricinus, Dermacentor reticulatus, and Haemaphysalis punctata. Eight patients developed local skin lesions at the site of the tick bite including non-specific lesions, itching sensation at the lesion site, and eschar. Most common TBPs detected in platelet fraction were Borrelia spielmanii and Rickettsia sp., followed by Borrelia afzelii and Anaplasma phagocytophilum. Multiple infections with three TBPs were detected in platelet fraction. In whole blood, most common TBPs detected were Anaplasma spp. and A. phagocytophilum, followed by Rickettsia spp. and B. afzelii. In ticks, the most common TBP detected was Rickettsia spp., followed by Borrelia spp. and Anaplasma spp. Overall, nine different pathogens with variable prevalence were identified using species-specific primers, and the most common was Rickettsia helvetica. In three patients, there were no coincidences between the TBPs detected in whole blood and tick samples. Only in one patient was detected A. phagocytophilum in both, whole blood and tick samples. These results suggest the unequal detection of TBPs in whole blood, platelet fraction and ticks collected, from the same individual. The results justify the use of both whole blood and platelet fraction for molecular diagnosis of TBPs in patients.

Mediterranean spotted fever-like illness (MSF-like illness) is a tick-borne disease caused by

Myiasis is a condition characterized by fly larvae infestation, most common in tropical regions, but with a risk of occurring anywhere in the world. Here, we report a case of nasal myiasis caused by a sarcophagid fly in a critically ill COVID-19 patient hospitalized in a reallocated ICU department in Serbia and discuss procedures that could prevent the occurrence of similar incidents in reallocated ICU departments worldwide.

Here we present the first case of Aerococcus urinae isolated from urine sample of a 5-year-old boy from Vojvodina province (Serbia). The patient had predisposing factors related to the susceptibility of genitourinary system to infection. Thus, he reported no complaints related to active urinary infections. Eradication of infection was achieved after 10-day treatment with meropenem. From this case, the question emerges: Is there a possibility that A. urinae infections are neglected in pediatric patients with susceptibility of genitourinary system to infection? Only after wide introduction of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) in competent laboratories, we will be able to give a precise answer.

Objective. To test if the direct immunofluorescence can be used for the detection of Borrelia afzelii in brain tissue during the standardization of the animal model of neuroborreliosis in NMRI mice. Methods. The study was performed on 15 mice of NMRI strain. All mice were subcutaneously inoculated with 100 ml of BSK-H medium containing the local isolate of Borrelia afzelii. Animals were sacrificed after inoculation at III (n = 4), IV (n = 6) and V (n = 5) weeks, by cervical dislocation. In the sampled brains of mice, the presence of Borrelia was detected by direct immunofluorescence (DIF) and chain polymerization reaction (PCR). Results. The first brain tested positive for Borrelia three weeks after the inoculation. The bacteria were detected in 1 out of 4 brains (25%). After that, there was a growth in the percentage of positive results. The data showed that 3 out of 6 brains (50%) were found positive on Borrelia presence by the end of the fourth week. Whereas, in 3 out of 5 brains (60%) Borrelia was detected five weeks following the inoculation. Conclusion. According to the preliminary results, direct immunofluorescence appeared to be a practical, low budget method for following the kinetics of neuro-infection. NMRI mice could be considered as an adequate animal model for neuroborreliosis. Thus, more research is needed on the topics of infection kinetics for the period after fifth week post inoculation, as well as sensitivity and specificity of direct immunofluorescence.

Introduction: Stress represents a set of reactions in the organism activated by external factors. In order to maintain homeostasis and protect the organism, numerous mechanisms for adaptation to stress evolved. Stressors that act in short-term period cause acute stress reaction with generally positive effect on organism. When the stressor persists, and the organism fails to respond to the challenge, chronic stress develops, leading to pathological conditions, such as women's menstrual cycle disorders. Aim: To examine the impact of chronic restraint stress on the estrous cycle in NMRI female mice. Material and methods: A number of 12 mature female NMRI mice were randomly divided into control (n = 6) and experimental (n = 6) group. The induction of stress was performed for the experimental group by using restrain chambers 2 hours daily in 14 days. From the beginning of the experiment, vaginal lavages were taken from all mice for making smears that were analyzed to determine estrous cycle stages. For analysis of chronic stress effect, the frequency of estrus stages alternation observed in experimental group compared to control was examined. Results: The estrous cycle was observed and divided into proestrus, estrus, metestrus and diestrus. During the adaptation period, prolonged diestrus was dominantly present in both groups. When stress was induced, in the experimental group animals the absence of diestrus stage and oscillation to other stages was obseved, in contrast to the control group, where the diestus stage was frequently observed. Frequency of diestrus stage deviation in stressed animals was shown to be statistically significant (p < 0.01) when compared to control. Conclusion: The induction of chronic restraint stress in female NMRI mice leads to the alternation of the estrous cycle. Considering the fact that NMRI female mice share the similar hormonal regulation of the estrous cycle with women's menstrual cycle, they could present a solid model for studying women's reproductive disorders.

The microflora of the gastrointestinal tract is essential for mucosal protection, immune education, and for metabolism of humans. Disturbances of these processes results in various diseases. Regarding these processes, there is a clinical interest in the utility of microbial therapy - the feeding of non-pathogenic bacteria, originally derived from the alimentary tract. Considering the commercial success of probiotics, there is still a difficulty for consumers as well as for physicians when choosing the specific product. Since they are classified as food supplements, and not subject to drug regulations, there is ongoing skepticism about the benefits they offer. Fecal bacteriotherapy is reported as highly effective in the treatment of various diseases, including those of the digestive tract. The treatment itself consists of feces infusion from a healthy human donor to the gastrointestinal tract of patient. According to current data, fecal bacteriotherapy represents an emerging and promising low-cost solution for diseases with which antibiotic and probiotic products have been struggling for years.

The reduction in the risk of transfusion-transmissible infections (including HIV infection) is an essential part of transfusion patient care. Here, we report the first incident of HIV transmission via transfusion in north Serbia due to blood donor dishonesty, and the failure of the laboratory screen tests to detect the presence of HIV particles in his blood. Infected blood products were distributed to two recipients, and HIV infection was confirmed in one. This incident finally led to the implementation of Nucleic Acid Amplification Technology as mandatory testing of blood donors for HIV infection in Serbia and raised many questions related to the responsibility and ethics of all the participants in the blood transfusion supply chain. There is a need for the implementation of modern and non-discriminative laws in Serbia in order to reduce transfusion-transmissible infections. In addition, transfusion institutes in Serbia need to be obliged to pursue the constant upgrade of their diagnostic capacities in order to prevent similar incidents and to provide the best possible care for blood donation recipients.

Chronic stress is regarded as a significant factor in the etiology of the many diseases. Numerous methods have been developed through which the effect of chronic stress is examined. The aim of this study is to demonstrate the new experimental model for analysis of immuno-suppression induced by chronic restraint stress, through challenge with conditionally tumorigenic cell line BHK-21/C13. 20 male NMRI mice were randomly divided into 2 groups-control and experimental. Each mouse was subcutaneously inoculated with BHK-21/C13 cells. Stress in the experimental group was induced for 20 days. After the experiment, tumor masses were removed, and analyzed using histology and immunohistochemistry techniques. We found a statistically significant difference (p = 0.034) in tumor expression and tumor volumes (p = 0.0061) between groups, as well as in immunopositivity on Ki67, cytochrome C and matrix metalloproteinase 9. Absence of immune infiltrate was noticed in experimental, and the presence of inflammatory infiltrate at tumor invasion front in control group.

The impact of tick-borne pathogens (TBPs) on human health has increased in the last decades, since the incidence of emerging and re-emerging infectious and zoonotic tick-borne diseases has increased worldwide. Tick-borne rickettsiae of the Spotted Fever group (SFGR) are considered as emerging pathogens that can infect humans and cause a variety of non-specific clinical symptoms. Here, we report nine cases of atypical tick-borne diseases (9/460; 1.95%) that occurred over a period of four months (from 15 April 2021 to 16 August 2021) in Serbia, from which five cases were classified as confirmed SFGR infection, two cases as probable SFGR infection and two cases as suspected SFGR infection. Within cases of confirmed SFGR infection, R. helvetica was detected as the causative agent in two cases. The most common clinical finding was non-expanding persistent circular redness, followed by eschar and enlargement of regional lymph nodes, and pain at lesion site. Rickettsia outer membrane protein B (ompB) and citrate synthase (gltA) gene fragments were amplified from clinical samples and ticks attached to patients and IgG reacting with Rickettsia conorii antigen were detected in sera samples of patients, which are highly suggestive of exposure to SFGR. Surveillance and monitoring of rickettsial diseases in Serbia should continue and extended to new areas due to the increasing trend of clinical infections caused by SFGR in the country.

Passive immunotherapy with convalescent COVID-19 plasma (CCP) is used as a therapeutic procedure in many countries, including Serbia. In this study, we analyzed the association between demographic factors, COVID-19 severity and the reactivity of anti-SARS-CoV-2 antibodies (Abs) in Serbian CCP donors. Individuals (n = 468) recovered from confirmed SARS-CoV-2 infection, and who were willing to donate their plasma for passive immunization of COVID-19 patients were enrolled in the study. Plasma samples were tested for the presence of IgG reactive to SARS-CoV-2 spike glycoprotein (S1) and nucleocapsid antigens. Individuals were characterized according to age, gender, comorbidities, COVID-19 severity, ABO blood type and RhD factor. Total of 420 candidates (420/468; 89.74%) reached the levels of anti-SARS-CoV-2 IgG that qualified them for inclusion in CCP donation program. Further statistical analysis showed that male individuals (p = 0.034), older age groups (p < 0.001), existence of hypertension (p = 0.008), and severe COVID-19 (p = 0.000) are linked with higher levels of anti-SARS-CoV-2 Abs. These findings will guide the selection of CCP donors in Serbia. Further studies need to be conducted to assess the neutralization potency and clinical efficiency of CCP collected from Serbian donors with high anti-SARS-CoV-2 IgG reactivity.

Aeromonas hydrophila is representative of group within the Aeromonadaceae family that mainly causes infections in humans. Aeromonads can induce meningitis, septicemia, respiratory and hemolytic uremic syndrome as well as gastroenteritis. Regarding diverse mechanisms involved in virulence and metabolic adaptation of Aeromonas hydrophila to various hosts and environments it is often introduced as ?Jack of all trades.? Here we report the first isolation of Aeromonas hydrophila from urine samples of three patients from the Urology Department within the Clinical Centre of Vojvodina. Epidemiological survey identified contact with surface water as the only mutual risk parameter. Following that, novel antibacterial agents against Aeromonas hydrophila are discussed

Mast cells are mainly distributed in tissues exposed to the external environment, but they are also found in ovaries, oviducts and uterus. We determined the difference in the number and distribution of Alcian Blue (+) and Safranine O (+) mast cells in reproductive systems of gravid and non-gravid female mice by histological examination. We showed that the number of mast cells is significantly higher in the reproductive system of gravid mice than in the reproductive system of non-gravid mice. Mast cells are unevenly distributed in ovaries, oviducts and uterus in both gravid and non-gravid mice. A large number of mast cells reside close to blood vessels throughout the entire reproductive system.

The majority of suggested mechanisms of Borrelia spreading inside erythema migrans (EM) are developed from in vitro studies and animal models. This report is the first to describe pathomorphological substrate of EM caused by Borrelia spielmanii in humans, addressing the hypothesis of enhanced Borrelia penetration through extracellular matrix. In the process of ruling out of atypical Masters’ disease, we conducted a punch biopsy of suspected EM and a two-tier serology testing for Lyme borreliosis, where we registered antibodies against B. spielmanii. Skin biopsy showed CD4+and CD8+lymphocyte involvement and high activity of matrix metalloproteinase 9. No alterations were detected in distribution and morphology of collagen type I and IV. Therefore, it is suggested that other mechanisms should be considered as major contributing factors to local spreading of B. spielmanii.

Introduction: A broad spectrum of research involving stress and stress protocols has long proven that a point of anhedonia, social defeat and learned helplessness can be achieved and observed – with anhedonia being a clinical symptom of Anxiety, Depression and Bipolar Affective Disorder. Aim: The aim of this study is the development of a simplified protocol for anhedonia induction in NMRI male mice in order to shorten the period of mice suffering and decrease complexity of the procedure for other researchers and introduction of new parameter in order to achieve better standardization of results. Materials and Methods: 21 male NMRI mice were introduced to 2 different stress protocols (one found in literature and one simplified) where cognitive- behavioral status was tested using the Sucrose Preference Test, Open Field Test, Grooming Pattern and histological examination of adrenal glands, and to propose a new protocol for fellow researchers. Results: Results observed include the successful induction of anhedonia proven by Sucrose Preference Tests, Barbering effect and microhemorrhage of the adrenal glands. Simplified protocol showed superiority compared to the one found in literature. Conclusion: Simplified protocol showed higher efficiency and reduced amount of work during testing phase. Introduction of NMI as a new parameter during behavioral evaluation resulted in better standardization of measured SPT values that incorporates common knowledge of mammal physiology. A big developmental step was the introduction of the Normalised Mass Index to even out mass fluctuations and differences in basal metabolism, which we recommend to other researchers and institutions.

Exiguobacterium aurantiacum is isolated from a variety of environmental samples but rarely from patients. The aim of the study was to represent isolation of unusual bacterial strains that could cause infection in patients. Final identification was performed using matrix-assisted description/ionization time-of-flight mass spectrometry (MALDI-TOF). Two isolates strains of E. aurantiacum were isolated, one isolate from distilled water used during surgical treatment and the second one from a patient with bacteremia after radical prostatectomy, both sensitive to all tested antimicrobials. Environmental strains could cause infection, especially in immunocompromised patients; therefore, rare bacteria testing is required, in which identification special assistance is provided by an automated system MALDI-TOF.

Lyme borreliosis (LB) is the most common tick-borne disease in Serbia and other European countries. Rabies is a fatal zoonosis distributed worldwide and is caused by the rabies virus. Professionals at risk of rabies—including veterinarians, hunters, communal service workers, and forestry workers—overlap with some professions at a higher risk of exposure to tick bites and tick-borne pathogen infections. We hypothesized that individuals identified by the public health system as at risk of rabies virus infection, and consequently vaccinated against rabies virus, also share a higher likelihood of Borrelia exposure. To test our hypothesis, a case-control study was carried out during 2019 in Serbia to determine the seroprevalence of anti-Borrelia antibodies in two case groups (individuals at risk and vaccinated against rabies virus) and a control group (individuals without risk of rabies). Individuals vaccinated against rabies following either “pre-exposure protocol” (PrEP, n = 58) or “post-exposure protocol” (PEP, n = 42) were considered as rabies risk groups and healthy blood donors (n = 30) as the control group. The results showed higher Borrelia seroprevalence in PrEP (17.2%, 10/58) and PEP (19.0%, 8/42) groups compared with the control group (6.67%, 2/30). Furthermore, odds ratio (OR) analysis showed that risk of rabies (in either the PrEP (OR = 2.91) or PEP (OR = 3.29) groups) is associated with increased odds of being seropositive to Borrelia. However, the difference in Borrelia seroprevalence between groups was not statistically significant (Chi-square (χ²) test p &gt, 0.05). The shared odds of LB and rabies exposure found in this study suggest that, in countries where both diseases occur, the common citizen can be at risk of both diseases when in a risky habitat. These findings are important to guide physicians in targeting high-risk groups, and diagnose LB, and to guide decision-makers in targeting control and prevention measures for both infections in risk areas.

The majority of suggested mechanisms of

Implementation of IMRT offers possibility to escalate radiation therapy dose without increased acute and late toxicity. The aim of this study is to compare acute and late genitourinary and gastrointestinal toxicity in patients treated with IMRT and 3DCRT technique. This study included 35 patients in study group a treated with IMRT technique, and 35 patients in study group B treated with 3DCRT technique. Patients were selected and referred to radical radiotherapy treatment prostate cancer. Acute genitourinary and gastrointestinal toxicity was evaluated during radiotherapy treatment according to recommendation of RTOG group. Late gastrointestinal and genitourinary toxicity was evaluated during regular control exams after radical radiotherapy treatment for six months. Based on the results χ2 test there was no statistical significant difference (p>0,05) between study group a i B in terms of acute gastrointestinal and genitourinary despite escalated radiotherapy dose in study group B treated with IMRT technique. Based on the results χ2 test there was no statistical significant difference (p>0, 05) between study group a i B in terms of late gastrointestinal and genitourinary toxicity. Intensity modulated radiation therapy is optimal technique in the radical treatment prostate cancer. This technique allows clinical benefit compared with 3D conformal radiotherapy-escalation of radiotherapy dose without increased toxicity in patients treated with IMRT technique.

Aim To characterize baby hamster kidney fibroblast (BHK 21/C13) cells and test the effects of antibodies against podoplanin and disodium cromolyn on BHK 21/C13 cell line-derived tumors grown on chick embryo chorioallantoic membrane (CAM). Material and methods BHK 21/C13 cell-derived fibrosarcomas developed in hamsters were implanted on CAM and treated with anti-podoplanin antibodies and disodium cromolyn. BHK 21/C13 cell immunophenotype was assessed. Results Fibrosarcoma cells were positive for vimentin, CD117, smooth muscle actin, vascular endothelial growth factor epidermal growth factor receptor, homebox prospero gene 1 and negative for platelet-derived growth factor B, neuron-specific enolase, S100, CD34, Ewing sarcoma and podoplanin. CAM-grown fibrosarcomas were highly sensitive to disodium cromolyn and anti-podoplanin antibodies. Conclusion Immunophenotyping BHK 21/C13 cells and their response to drugs represent the first step in revealing cell line utility and a reliable tool for experimental cancer research.

Although clinical trials have not been completed, it has already been confirmed that mebendazole, a well-known anti-parasitic drug widely used in the tropical areas, inhibits cancer cell growth. Preclinical studies show that mebendazole notably impedes the growth of malignant and metastatic tumors such as osteosarcoma and soft tissue sarcoma, melanoma, carcinoma (lung, colorectal, breast, ovarian, hepatocellular and adrenocortical), acute myeloid leukaemia, glioblastoma multiforme and meduloblastoma. Mebendazole can induce the depolymerization of microtubules in neoplasms and newly formed vasculature, stopping tumor growth and neoangiogenesis, along with other proposed mechanisms of action.Keywords: Anthelmintic, Mebendazole, Cancer treatment, Antimicrotubullar effect, Antineoangiogenesis

International audience; Tick-borne pathogens (TBPs) pose a major threat to human health in Europe and the whole northern hemisphere. Despite a high prevalence of TBPs in Ixodes ricinus ticks, knowledge on the incidence of tick-borne diseases in humans infested by this tick species is limited. This study was conducted in the year 2019 on patients who presented themselves to the Pasteur Institute Novi Sad with tick infestations. Ticks (n = 31) feeding on human (n = 30) and blood samples from the same individuals were collected by physicians and a microfluidic real-time high-throughput PCR system was used to test the genomic DNA of the samples for the presence of 27 bacterial and eight parasitic microorganisms in Serbia. Except for one Rhipicephalus sanguineus s.l. adult male tick, all ticks infesting humans were morphologically identified as I. ricinus. A high proportion of ticks (74 %, 23/31) were infected with at least one of the tested TB microorganisms, being Rickettsia helvetica (54 %, 17/31) the most common pathogen, but Borrelia afzelii (9 %, 3/31), Anaplasma phagocytophilum (6 %, 2/31), Borrelia miyamotoi (6 %, 2/31), and Francisella like-endosymbiont (6 %, 2/31), Borrelia valaisiana (3 %, 1/31), Borrelia lusitaniae (3 %, 1/31), Rickettsia felis (3 %, 1/31) and Rickettsia aeschlimannii (3 %, 1/31) were also identified. Despite the high infection rate of TBPs in ticks, only two human blood samples (6 %, 2/30) tested positive for the presence of TBPs, one patient (code H12, 67 years old female) was diagnosed with Borrelia spp. and the other patient was diagnosed (code H17, 71 years old female) with R. felis infection. The tick infesting patient H12 tested positive for B. afzelii, and R. helvetica and the tick infesting patient H17 tested positive for R. felis. Upon clinical examination, both patients were diagnosed with erythema migrans. No additional discomfort was reported by the patient and no additional pathology was observed by the physician. We concluded that humans bitten by I. ricinus in Serbia are exposed to a diverse array of TBPs with clinical impact in the Serbian cohort studied.

Occipital condyles, located at the inferior sides of lateral parts of occipital bone, are important structures that connect the cranium and the vertebral column. Their size, shape, location and, also, their congruence with superior articular facets of the atlas vertebra are of great importance for the stability of craniovertebral junctions. The progress in medical diagnostic and surgical techniques in the area of foramen magnum, requires the knowledge regarding anatomical aspects of this region. The purpose of this study was to evaluate the measurements of the occipital condyles, as well as to analyse the variations in the shape of the condyles and their position in relation to the foramen magnum. This study included 25 adult human skulls (11 male and 14 female) from the Osteological collection of the Department of Anatomy at Medical Faculty in Novi Sad. Nine parameters, which define morfological properties of the occipital condyles, were measured using Vernier caliper. Also, the shape and location of the condyles, as well as the narrowness of the foramen magnum were described. Our findings show that length of the left occipital condyle, as well as distance between the posterior top of the left occipital condyle and basion are statistically greater in male crania, comparing to female crania. According to the shape of the occipital condyles, we classified them in seven groups. Predominant type is oval shaped. The condylar foramen is bilaterally present in 36 % and the occipital condyles bilaterally protrude the foramen magnum in 40 % of analyzed crania. Morphometric analysis of the occipital condyles showed variations in their size, shape, presence of condylar canal and relation to the foramen magnum.

From the beginning of the 20th Century, scientific interest for animal tumor models has continuously grown. Early works confirmed tumorigenic effect of many animal viruses, parasites, chemicals like tar, smoke and many others. Influences of animal species and their immune status have also a great impact. Our laboratory at the Faculty of Medicine in Novi Sad introduced tumor models in teaching process for students on the bachelor degree and PhD level. We used models as spontaneous tumors in laboratory animals, chemically induced tumors, and transplanted tumors for demonstration of drug effects and some immune mechanisms in tumor biology. From spontaneous tumors we examined micemammary carcinoma, mice lymphoma, hedgehog ovarian carcinoma, and dog prostate hyperplasia. For chemically induction of neoplasia we used 4-NQO in mice and produced squamous carcinoma of the tongue. In the group of transplanted tumors, we used Ehrlich carcinoma in mice and BHK 21C13 sarcoma in diverse species of hamsters and mice. Tested drugs were thalidomide, dexamethason, heparin, metformin, doxorubicin, podoplanin. At this moment, examination of favorable effects of immobilization stress on tumor growth is in progress. Spontaneous and chemically induced tumors need time for growth and their appearance is rare. Because of that, formation of experimental groups is very complicated. We conclude that model of BHK 21C13 cell culture induced sarcoma in Syrian hamsters is the best model, easy reproducible and without influence of host immune mechanisms.