Morphological and functional changes in rats` skin following epicutaneous application of warfarin

Varfarin (4-OH kumarin) je antikoagulant prve generacije koji je u upotrebi kao lek i rodenticid. Osnovni biološki efekat ostvaruje inhibicijom vitamin K-zavisnog puta u kompletnoj sintezi brojnih faktora koagulacije u jetri. Varfarin deluje i na druge proteine za čiju je biološku aktivnost neophodan vitamin K, kao što su proteini uključeni u proces rasta, kalcifikacije kostiju, proliferacije mezangijskih ćelija, i proteine uključene u prenos ćelijskih signala. Širokom upotrebom varfarina kao leka i kao rodenticida uočeni su njegovi brojni neželjeni efekti. U terapijskim dozama on može da uzrokuje akutna i hronična neželjena dejstva, kao što su nekroza kože i kalcifikacija zida arterija, a osim antikoagulantnog može da ima i proinflamatorno i antiinflamatorno sistemsko dejstvo. Nepravilno rukovanje u proizvodnji i upotrebi varfarina kao rodetnicida može dovesti do jednokratne ili ponavljane inhalacione, oralne ili perkutane ekspozicije. Kontakt kože sa antikoagulantnim rodenticidima može da uzrokuje perkutanu intoksikaciju sa razvojem koagulopatije i krvarenja čak do smrtnog ishoda i lokalnu iritaciju kože. Poseban problem predstavlja neprepoznata izloženost malim dozama varfarina čiji se efekti mogu manifestovati i kao simptomi nepoznate bolesti i tada se varfarin može posmatrati i kao ksenobiotik. U našim eksperimentalnim istraživanjima opredelili smo se za ispitivanje ksenobiotičkih efekata varfarina u koži, s obzirom da je ona čest put ekspozicije, a ovakvi njegovi efekti nisu poznati U početnim istraživanjima utvrdili smo da epikutano aplikovan varfarin pokazuje lokalni i sistemski proinflamatorni, prooksidativni i imunomodulatorni efekat, a zavisno od aplikovane doze i direktan imunotoksični i dermatotoksični efekat. Preliminarna histomorfološka analiza je pokazala promene u koži koje mogu, sa jedne strane, da odražavaju inflamatorno-reparatorni program, ali, sa druge, da predstavljaju uvod u patološke događaje u koži. U ovom istraživanju ispitan je efekat jednokratne lokalne epikutane aplikacije varfarina na koži kod Dark Agouty pacova u dozama od 1μg, 10μg i 100μg. Analizirana je sedmodnevna dinamika selektivnih funkcionalnih parametara oksidativnog stresa, promena u imunskom sistemu, mikrovaskulaturi, ćelijskom ciklusu keratinocita i dermalnih fibroblasta i strukturne promene u koži histohemijskim, imunohistohemijskim metodama i morfometrijskim analizama uz pomoć svetlosnog, elektronskog i konfokalnog mikroskopa...Warfarin (4-OH coumarin) is an anticoagulant of the first generation which is used as a drug and rodenticide. It’s main biological effect is achieved through inhibition of vitamin K-dependent path in a complete synthesis of many coagulation factors in the liver. Warfarin acts on proteins for which biological activity of vitamin K is necessary, such as proteins involved in the process of growth, bone calcification, proliferation of mesangial cells, and proteins involved in the transduction of cellular signals. By widespread use of warfarin as a drug and as a rodenticide, it’s numerous side effects have been noted. At therapeutic doses, it may cause acute and chronic side effects, such as skin necrosis and calcification of the arterial wall, and, besides anticoagulant, it can have proinflammatory and anti-inflammatory systemic effect. Improper handling in the production and use of warfarin as rodetnicide can lead to a single or repeated inhalational, oral or percutaneous exposure. Contact between skin and anticoagulant rodenticides can cause percutaneous intoxication with the development of coagulopathy and bleeding, death and local skin irritation. A particular problem presents unrecognized exposure to low doses of warfarin whose effects can be manifested as symptoms of an unknown disease and then warfarin can be seen as a xenobiotic. In our experimental study, we decided to examine the xenobiotic effects of warfarin in the skin, since it is a common route of exposure, and such effects of warfarin are not known. In initial studies we have found that epicutaneously applied warfarin show local and systemic proinflammatory, pro-oxidative and immunomodulatory effects and, depending on the applied dose, direct immunotoxic and dermatotoxic effect. Preliminary histomorphological analysis revealed changes in the skin that can, on the one hand, reflect the inflammatory-reparative program, but, on the other, can represent an introduction to the pathological events in the skin. In this study the effect of a single local epicutaneous application of warfarin on the skin of Dark Agouty rats at doses of 1μg, 10μg and 100μg was examined. The seven-day dynamics of selective functional parameters of oxidative stress, changes in the immune system, microvasculature, the cell cycle of keratinocytes and dermal fibroblasts and structural changes in the skin, have been analyzed by immunohistochemical methods and morphometric analysis with the help of light-, electron- and confocal microscope..

Fulminant Wegener's granulomatosis: A case report

Introduction. Granulomatosis Wegener is anti-neutrophil cytoplasmic antibodies (ANCAs)-associated systemic vasculitis of unknown etiology. It is manifested as granulomatous necrotizing inflammation of the upper and lower parts of the respiratory tract, glomerulonephritis and systemic vasculitis involving most frequently the skin and oral mucous membrane. Sera markers of this disease are c-ANCA and p-ANCA. Case report. We presented a female patient aged 52 years with purpuric spots that had appeared on the lower legs ten months before admission to our hospital. The disease ran an aggressive course, and a month before admission hemorrhagic bullae, skin ulcers, hoarseness, dyspnea, generalized arthralgia, fatigue and fever had rapidly developed. Histopathological examination of a skin sample revealed necrotizing vasculitis, so that sera markers concentrations were elevated (c-ANCA, p-ANCA). There was a perforation of the nasal septum found on rhinoscopy. During hospitalization acute abdominal pain occurred, a possible tumor in the small intestine and possible granulomas in the liver were seen by multislice computed tomography (MSCT) examination, with normal findings on the lungs and kidneys. The treatment started with methylprednisolone: 500 mg/d i.v. infusion for consecutive 3 days, then 60 mg/d. On exploratory laparotomy small bowel perforation and diffuse peritonitis were found. Unstable in the postoperative period, the patient died on the day 12 of hospitalization. Conclusion. The reported patient was with fulminant Wegener’s granulomatosis, dominantly with skin changes and with gastrointestinal manifestation. This case accents the need for rapid systemic clinical evaluation in a severely ill patient with unclear diagnosis

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

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

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

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

First record of Calodium hepaticum and Taenia taeniaeformis liver infection in wild Norway rats (Rattus norvegicus) in Serbia

The nematode Calodium hepaticum and the cestode Taenia taeniaeformis are zoonotic helminths primarly found in the liver of common wild rats. Most reports on these helminth species with cosmopolitan distribution are from Asia, and there is paucity of data for Europe. Wild Norway rats (Rattus norvegicus) from urban and suburban habitats of the Belgrade area were examined for the presence of Calodium hepaticum and Taenia taeniaeformis larvae liver infections. The presence of visible cysts and a histomorphology of parasite-related inflammatory liver responses were sought as signs of infection. The total prevalence of infection was 10.9% (C. hepaticum) and 29.9% (T. taeniaeformis), with no differences between the sexes. No difference in the annual prevalence of both helminth species was noted. Data obtained in this study provide new information relevant to wild Norway rats as sources of C. hepaticum and T. taeniaeformis liver infection in this geographic area, and, in a wider context, in Europe. .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

Morphological and functional changes in rats` skin following epicutaneous application of warfarin

Varfarin (4-OH kumarin) je antikoagulant prve generacije koji je u upotrebi kao lek i rodenticid. Osnovni biološki efekat ostvaruje inhibicijom vitamin K-zavisnog puta u kompletnoj sintezi brojnih faktora koagulacije u jetri. Varfarin deluje i na druge proteine za čiju je biološku aktivnost neophodan vitamin K, kao što su proteini uključeni u proces rasta, kalcifikacije kostiju, proliferacije mezangijskih ćelija, i proteine uključene u prenos ćelijskih signala. Širokom upotrebom varfarina kao leka i kao rodenticida uočeni su njegovi brojni neželjeni efekti. U terapijskim dozama on može da uzrokuje akutna i hronična neželjena dejstva, kao što su nekroza kože i kalcifikacija zida arterija, a osim antikoagulantnog može da ima i proinflamatorno i antiinflamatorno sistemsko dejstvo. Nepravilno rukovanje u proizvodnji i upotrebi varfarina kao rodetnicida može dovesti do jednokratne ili ponavljane inhalacione, oralne ili perkutane ekspozicije. Kontakt kože sa antikoagulantnim rodenticidima može da uzrokuje perkutanu intoksikaciju sa razvojem koagulopatije i krvarenja čak do smrtnog ishoda i lokalnu iritaciju kože. Poseban problem predstavlja neprepoznata izloženost malim dozama varfarina čiji se efekti mogu manifestovati i kao simptomi nepoznate bolesti i tada se varfarin može posmatrati i kao ksenobiotik. U našim eksperimentalnim istraživanjima opredelili smo se za ispitivanje ksenobiotičkih efekata varfarina u koži, s obzirom da je ona čest put ekspozicije, a ovakvi njegovi efekti nisu poznati U početnim istraživanjima utvrdili smo da epikutano aplikovan varfarin pokazuje lokalni i sistemski proinflamatorni, prooksidativni i imunomodulatorni efekat, a zavisno od aplikovane doze i direktan imunotoksični i dermatotoksični efekat. Preliminarna histomorfološka analiza je pokazala promene u koži koje mogu, sa jedne strane, da odražavaju inflamatorno-reparatorni program, ali, sa druge, da predstavljaju uvod u patološke događaje u koži. U ovom istraživanju ispitan je efekat jednokratne lokalne epikutane aplikacije varfarina na koži kod Dark Agouty pacova u dozama od 1μg, 10μg i 100μg. Analizirana je sedmodnevna dinamika selektivnih funkcionalnih parametara oksidativnog stresa, promena u imunskom sistemu, mikrovaskulaturi, ćelijskom ciklusu keratinocita i dermalnih fibroblasta i strukturne promene u koži histohemijskim, imunohistohemijskim metodama i morfometrijskim analizama uz pomoć svetlosnog, elektronskog i konfokalnog mikroskopa...Warfarin (4-OH coumarin) is an anticoagulant of the first generation which is used as a drug and rodenticide. It’s main biological effect is achieved through inhibition of vitamin K-dependent path in a complete synthesis of many coagulation factors in the liver. Warfarin acts on proteins for which biological activity of vitamin K is necessary, such as proteins involved in the process of growth, bone calcification, proliferation of mesangial cells, and proteins involved in the transduction of cellular signals. By widespread use of warfarin as a drug and as a rodenticide, it’s numerous side effects have been noted. At therapeutic doses, it may cause acute and chronic side effects, such as skin necrosis and calcification of the arterial wall, and, besides anticoagulant, it can have proinflammatory and anti-inflammatory systemic effect. Improper handling in the production and use of warfarin as rodetnicide can lead to a single or repeated inhalational, oral or percutaneous exposure. Contact between skin and anticoagulant rodenticides can cause percutaneous intoxication with the development of coagulopathy and bleeding, death and local skin irritation. A particular problem presents unrecognized exposure to low doses of warfarin whose effects can be manifested as symptoms of an unknown disease and then warfarin can be seen as a xenobiotic. In our experimental study, we decided to examine the xenobiotic effects of warfarin in the skin, since it is a common route of exposure, and such effects of warfarin are not known. In initial studies we have found that epicutaneously applied warfarin show local and systemic proinflammatory, pro-oxidative and immunomodulatory effects and, depending on the applied dose, direct immunotoxic and dermatotoxic effect. Preliminary histomorphological analysis revealed changes in the skin that can, on the one hand, reflect the inflammatory-reparative program, but, on the other, can represent an introduction to the pathological events in the skin. In this study the effect of a single local epicutaneous application of warfarin on the skin of Dark Agouty rats at doses of 1μg, 10μg and 100μg was examined. The seven-day dynamics of selective functional parameters of oxidative stress, changes in the immune system, microvasculature, the cell cycle of keratinocytes and dermal fibroblasts and structural changes in the skin, have been analyzed by immunohistochemical methods and morphometric analysis with the help of light-, electron- and confocal microscope..