Helminth fauna of Rattus norvegicus Berkenhout, 1769 from the Belgrade area, Serbia

The aims of this study were to provide baseline knowledge about intestinal parasites in Norway rats (Rattus norvegicus) that inhabit the Belgrade area, and to analyze the associations among helminths. Of 302 trapped rats, 52% were females and 48% males, with 39% and approx. 37% of juvenile-subadult individuals, per sex, respectively. The following parasites were detected (with their respective prevalence): Cestoda - Hymenolepis diminuta (30.46%) and Rodentolepis fraterna (12.58%); Nematoda - Heterakis spumosa (36.75%), Nippostrongylus brasiliensis (16.22%), Capillaria sp. (5.96%), Trichuris muris (5.96%), Syphacia muris (4.30%) and Strongylus sp. larvae (0.33%). Flukes (Trematoda) were not recorded. Of all examined rats, 68.54% were found to harbor at least one parasite species, with higher prevalence in male hosts and in adult individuals. There were no age-related differences in the prevalence of infection with individual helminth species. Multiple infections occurred with up to four species per rat showing different combinations of parasite infections. These are the first records of the gastrointestinal helminth fauna of Norway rats in Serbia.Projekat ministarstva br. 14303

Characteristics of local pulmonary response following intranasal application of Aspergillus fumigatus conidia

In this study, histopathology of local pulmonary response following intranasal inoculation of different doses of Aspergillus fumigatus conidia in laboratory rats was evaluated. Development of response was evaluated on days seven and twenty one, following conidia inoculation by cell infiltration and by presence of A. fumigatus conidia in homogenates of lung tissue. Total and differential peripheral blood leukocyte counts and state of leukocyte adhesion/aggregation were monitored to estimate the presence of systemic response in infected individuals. Mycological examination revealed the presence of conidia in lung tissue homogenates of infected animals, with high number of non-germinating spores on day twenty one, following the inoculation of lower conidia dose. Histopathological examination revealed the presence of lymphocytes perivascularly and in vascular lumen in the lungs. Presented data demonstrate pulmonary immune response following A. fumigatus conidia administration.Aspergillus fumigatus je oportunistička gljiva koja najčešće dovodi do pulmonarnih aspergiloza kod imunokompromitovanih organizama. U ovom radu su prikazani rezultati istraživanja izazivanja infekcije aspergilusom kod pacova. U cilju dobijanja podataka o prisustvu i karakteristikama lokalnog inflamatornog odgovora u plućima određen je diferencijalni sastav leukocita periferne krvi i testirana je njihova aktivnost, izvršeni su histopatološki pregled pluća i provera prisustva spora A. fumigatus u homogenatima pluća. Naši podaci su pokazali prisustvo lokalnog odgovora u plućima posle 7 dana dok podaci dobijeni posle 21 dan ne pokazuju značajne razlike u odnosu na kontrolnu grupu. Povećani broj neklijajućih spora u homogenatu pluća sedmog dana nakon inokulacije može da se objasni prisustvom limfocitnog infiltrata u plućima i aktivnošću limfocita in situ. Povećan broj klijajućih spora 21. dana nakon inokulacije može da bude posledica kapaciteta malog broja preostalih spora da klijaju.Projekat ministarstva br. 14303

Characteristics of local pulmonary response following intranasal application of Aspergillus fumigatus conidia

In this study, histopathology of local pulmonary response following intranasal inoculation of different doses of Aspergillus fumigatus conidia in laboratory rats was evaluated. Development of response was evaluated on days seven and twenty one, following conidia inoculation by cell infiltration and by presence of A. fumigatus conidia in homogenates of lung tissue. Total and differential peripheral blood leukocyte counts and state of leukocyte adhesion/aggregation were monitored to estimate the presence of systemic response in infected individuals. Mycological examination revealed the presence of conidia in lung tissue homogenates of infected animals, with high number of non-germinating spores on day twenty one, following the inoculation of lower conidia dose. Histopathological examination revealed the presence of lymphocytes perivascularly and in vascular lumen in the lungs. Presented data demonstrate pulmonary immune response following A. fumigatus conidia administration

Basic indices of spleen immune activity in natural populations of Norway rats (Rattus norvegicus berkenhout, 1769) in Serbia

Basic parameters of spleen immune activity (spleen weight, histomorphology of splenic compartments, and mitogen-induced splenocyte proliferative capacity in vitro) were evaluated in adult individuals of wild Norway rats from urban habitats and compared to the same data obtained in laboratory rat strains. A wider range of relative spleen mass and differential histomorphological characteristics, together with differences in the level and pattern of responsiveness of splenocytes to exogenous stimulation, were noted in spleens of wild Norway rats. Evidence of both enhanced and low-level immune-relevant spleen activity in wild rats demonstrates the complexity of changes in spleen immune activity in rats from natural populations.Ispitivani su osnovni parametri imunske aktivnosti u slezini (masa slezine, histomorfologija i sposobnost ćelija slezine da profilerišu u odgovoru na mitogen u in vitro uslovima) kod adultnih jedinki sivog pacova iz urbanih staništa i poređeni sa podacima dobijenim kod laboratorijskih sojeva pacova. Kod jedinki iz prirodnih populacija je zapažen veći opseg relativnih masa slezine, različite histomorfološke karakteristike i razlike u nivou i načinu odgovara ćelija slezine na egzogenu stimulaciju u poređenju sa jedinkama laboratorijskih sojeva. Ovi podaci ukazuju na kompleksne promene u imunskoj aktivnosti slezine kod jedinki iz prirodnih populacija.Projekat ministarstva br. 14303

Production of TNF-alpha by skin explants of dinitrochlorobenzene-challenged ears in rats: A model for the evaluation of contact hypersensitivity

Background. Contact hypersensitivity (CHS) is a local inflammatory response of the skin following challenge of hapten-sensitized animals. It is the consequence of cell infiltration of derm and the release of inflammation mediators, among which Tumor necrosis factor-alpha (TNF-α) is one of the most important factors. The intensity of the inflammation could be quantified by ear swelling which is the classical manifestation of the reaction. This study was testing the working hypothesis that levels of TNF-α in skin organ culture medium should correlate with the intensity of CHS reaction measured in vivo by ear swelling assay, and with the density of dermal infiltrate in ear skin samples. In order to test the working hypothesis, the intensity of inflammatory reaction following challenge was evaluated by classical measurements of ear swelling, by the determination of TNF-α levels in culture fluids of ear skin following epicutaneous application of dinitrochlorobenzene (DNCB) into the ears of sensitized animals. Methods. Animal model of CHS reaction to DNCB in Albino Oxford rats was used as described. Ear swelling was quantified in percentage terms as the difference in thickness between the challenged and nontreated ears of the same animal. Dermal infiltrate density in histopathologically analyzed samples of ear skin was evaluated by computer-assisted image analysis. Ear skin samples were cultured in standard medium for 24 h, and TNF-α concentration in the conditioned medium was subsequently determined with ELISA test. Results. Dose-dependent increase in the density of the dermal infiltrate and in TNF-α in CM were noted following the application of 0.65%, 1.3% and 2.6% of DNCB to the ears of previously sensitized rats. The correlation between ear swelling and the levels of TNF-α (r=0.933, p<0.001) in CM, and between ear swelling and dermal infiltrate density (r=0.916, p<0.001) was found. Correlation was also found between the density of the dermal infiltrate and the levels of TNF-α (r=0.865, p<0.001). Conclusion. Presented data suggested that skin-organ culture system and the quantification of inflammatory mediators might be used for the evaluation of contact hypersensitivity reaction and its intensity

Immune-related health-relevant changes in natural populations of Norway rat (Rattus norvegicus Berkenhout, 1769): White blood cell counts, leukocyte activity, and peripheral organ infiltration

Basic immune-related health-relevant changes (total and differential white blood cell counts and activity, leukocyte tissue infiltration, and related pathohistology) were assessed in wild Norway rats from urban habitats. Comparative measurements were conducted in individuals of several laboratory strains of Norway rat in order to gain insight into environmental effects on the health of wild rats. Changes in leukocyte counts and activity along with tissue infiltration were noted only in wild rats, indicating systemic as well as tissue inflammation in these animals. Coincidence of these changes with chronic inflammatory pulmonary and kidney disease was observed in the majority of affected rats.U radu su ispitane osnovne imunološke zdravstveno značajne promene kod jedinki sivog pacova iz urbanih staništa (ukupan broj i diferencijalni sastav, kao i aktivnost leukocita periferne krvi, tkivna infiltracija leukocita i patohistološke promene). Uporedo su rađena ispitivanja na nekoliko laboratorijskih sojeva pacova da bi se stekao uvid u efekte spoljašnje sredine na zdravlje jedinki iz prirodnih populacija. Promene u broju i aktivnosti leukocita, kao i infiltracija u organe su primećene samo kod jedinki iz prirodnih populacija i ukazuju na sistemsku i tkivnu inflamaciju kod tih jedinki. Kod većine obolelih jedinki je pokazana povezanost ovih promena i hroničnih inflamatornih oboljenja pluća i bubrega.Projekat ministarstva br. 14303

Helminth fauna of Mus musculus Linnaeus, 1758 from the suburban area of Belgrade, Serbia

The helminth fauna of the house mouse (Mus musculus Linnaeus, 1758) was studied on the basis of 429 host individuals from the suburban area of Belgrade. Eleven helminth species were recorded: three cestode species - Catenotaenia pusilla, Rodentolepis fraterna, and Cysticercus (= Strobilocercus) fasciolaris [larval stage of Taenia taeniaeformis (Batsch, 1821)]; and eight nematode species - Heligmosomoides polygyrus, Syphacia sp., Aspiculuris tetraptera, Syphacia obvelata, Heterakis spumosa, Trichuris muris, Mastophorus muris, and Gongylonema sp. Within the general helminth fauna, H. polygyrus was found to be the most prevalent species (39.2%) and caused the highest infection intensity. Prevalences of A. tetraptera, C. pusilla, and S. obvelata ranged from 12.8% to 6.1%, while the remaining species showed prevalences ranging from 4.9% (for Syphacia sp.) to 0.2% (for Gongylonema sp.). All the species found in males were also present in females, with the exceptions of M. muris and Gongylonema sp. No significant differences were found between males and females regarding prevalence (P%), mean infection intensity (MI), or mean abundance (MA).Helmintofauna kućnog miša (Mus musculus) istraživana je kod 429 jedinki izlovljenih sa suburbanog područja Beograda (selo Jabuka, 15 km severoistočno od Beograda), a prisustvo 3 vrste pantljičara (Cestoda) i 8 vrsta valjkastih crva (Nematoda) utvrđeno je u 262 jedinke (61.1%). Od pantljičara su bile prisutne Catenotaena pusilla, Rodentolepis fraterna i larveni stadijum Taenia taeniaeformis - Cysticercus (= Strobilocercus) fasciolaris, a od valjkastih crva Helygmosomoides polygirus, Syphacia sp., S. obvelata, Aspiculuris tetraptera, Heterakis spumosa, Trichuris muris, Mastophorus muris i Gongylonema sp. Dominantna vrsta bila je H. polygyrus sa prevalencom od 39,2% i prouzrokovala je najviši intenzitet infekcije. Za A. tetraptera, C. pusilla i S. obvelata prevalence su iznosile od 12,8 do 6,1%, a za ostale vrste helminata od 4,9 (za Syphacia sp.) do 0,2% (za Gongylonema sp.). Sve vrste nađene u mužjacima kućnog miša bile su nađene i u ženkama, izuzimajući M. muris i Gongylonema sp. Nisu nađene statistički značajne razlike između mužjaka i ženki kućnog miša u pogledu prevalence (P%) i srednjih vrednosti intenziteta infekcije (MI) i abundancije (MA).Projekat ministarstva br. 14303

Biodegradable polymer/hydrogel composite for controlled delivery of cationic formulations

Composites of biodegradable polymers and hydrogels are promising materials for controlled delivery systems with prolonged drug release. In this contribution, we present an innovative implant design comprising poly(DL-lactide-co-ε-caprolactone) copolymer base and a crosslinked poly(acrylic acid) hydrogel. Implants were prepared in the form of disks using the modified traditional liquid phase inversion process. Solutions containing all implant precursors were dispensed into transparent non-stick molds and cured by UV irradiation. UV curing was followed by immersion into the phosphate buffer solution bath to achieve phase separation and solidification. Structure and composition of the implant were characterized using SEM and FTIR. Obtained implants exhibited high loading capacity for cationic formulations and a moderate degree of swelling. Studies of implant loading and subsequent release of methylene blue into the phosphate-buffered saline demonstrated diffusioncontrolled delivery kinetics over a period of several weeks. To assess biocompatibility of implants as possible materials for drug delivery systems in mammals, we evaluated their effects on viability (Trypan blue exclusion assay), metabolic activity, proliferation (MTT assay) and priming (nitric oxide/NO production) of freshly isolated rat splenocytes during 24 h and 48 h of cultivation. The viability was unaltered, metabolic activity/proliferation was increased after 48 h and the decrease of NO production, as well as drop in responsiveness to cell mitogen concanavalin A (ConA) in cells on implants were observed. These results suggest that implants could be used as a suitable material for drug delivery systems, but their capacity to stimulate cell proliferation and their immunosuppressive potential deserve further investigations

Helminth fauna of Rattus norvegicus Berkenhout, 1769 from the Belgrade area, Serbia

The aims of this study were to provide baseline knowledge about intestinal parasites in Norway rats (Rattus norvegicus) that inhabit the Belgrade area, and to analyze the associations among helminths. Of 302 trapped rats, 52% were females and 48% males, with 39% and approx. 37% of juvenile-subadult individuals, per sex, respectively. The following parasites were detected (with their respective prevalence): Cestoda - Hymenolepis diminuta (30.46%) and Rodentolepis fraterna (12.58%); Nematoda - Heterakis spumosa (36.75%), Nippostrongylus brasiliensis (16.22%), Capillaria sp. (5.96%), Trichuris muris (5.96%), Syphacia muris (4.30%) and Strongylus sp. larvae (0.33%). Flukes (Trematoda) were not recorded. Of all examined rats, 68.54% were found to harbor at least one parasite species, with higher prevalence in male hosts and in adult individuals. There were no age-related differences in the prevalence of infection with individual helminth species. Multiple infections occurred with up to four species per rat showing different combinations of parasite infections. These are the first records of the gastrointestinal helminth fauna of Norway rats in Serbia.Projekat ministarstva br. 14303

Tumor necrosis factor-alfa and interleukin-4 in cerbrospinal fluid and plasma in different clinical forms of multiple sclerosis

Background/Aim. Multiple sclerosis (MS) is an immunemediated central nervous system disease characterized by inflammation, demyelination and axonal degeneration. Cytokines are proven mediators of immunological process in MS. The aim of this study was to investigate whether there is a difference in the production of the tumor necrosis factor alpha (TNF-alpha) and interleukin-4 (IL-4) in cerebrospinal fluid (CSF) and plasma in the MS patients and the controls (other neurological non-inflammatory diseases) and to determine a possible difference in these cytokines in plasma and CSF in different clinical forms of MS. Methods. This study involved 60 consecutive MS patients - 48 patients with relapsing-remitting MS (RRMS) and 12 patients with secondary progressive MS (SPMS). The control group consisted of 20, age and sex matched, nonimmunological, neurological patients. According to the clinical presentation of MS at the time of this investigation, 34 (56.7%) patients had relapse (RRMS), 14 (23.3%) were in remission (RRMS), while the rest of the patients, 12 (20.0%), were SPMS. TNF-alpha and IL-4 concentrations were measured in the same time in CSF and plasma in the MS patients and the controls. Extended disability status score (EDSS), albumin ratio and IgG index were determined in all MS patients. Results. The MS patients had significantly higher CSF and plasma levels of TNF-alpha than the controls (p < 0.001 for both samples). IL-4 CSF levels were significantly lower in the MS patients than in the controls (p < 0.001), however plasma levels were similar. The patients in relapse (RRMS) and with progressive disease (SPMS) had higher concentrations of CSF TNF-alpha levels than the patients in remission (p < 0.001). IL-4 CSF levels in relapse (RRMS) and SPMS groups were lower than in the patients in remission. The patients in remission had an unmeasurable plasma TNF-alpha level and the patients with SPMS had significantly lower IL-4 levels in plasma than the patients in relapse and remission (p < 0.001). The only significant correlation between cytokine level with either EDSS, or albumin ratio, or IgG index, was found between CSF TNF-alpha levels and albumin ratio in the patients with relapse (R square = 0.431, p < 0.001). Conclusion. According to the obtained data MS relapse was characterized by high concentrations of TNF-alpha in CSF and plasma and low concentrations of IL-4 in CSF. Remission was characterized by high concentrations of IL-4 and low concentrations of TNF-alpha both in CSF and plasma. SPMS was characterized with lower concentrations of TNF-alpha and IL-4 compared to relapse, both in CSF and plasma