Idiopatski i sekundarni stečeni megakolon kod pasa udruženi su sa smanjenom vip-inervacijom u oštećenom kolonu

It is well established that megacolon in carnivores, including both cats and dogs, is a common finding. Megacolon occurs more often in the cat that the dog. Based on current data idiopathic megacolon is a common cause of constipation in cats (62% of constipated cats are affected by diopathic megacolon). There is no evidence of idiopathic megacolon in dogs and publications about this disease in this species is very scarce. We investigated the enteric nervous system in the dilated portion (DP) of the colon in dogs with idiopathic aquired (n=7) or secondary aquired megacolon (n=21) and compared the results with a normal colon in control dogs (n=3). Colonic sections of surgical specimens were investigated by conventional and immunohistochemical methods, including pan-neuronal markers (NSE, synaptophisin, and neurofilament) and VIP, as well as S-100 protein for detection of ganglionic glial cells. Compared to controls, the two megacolon groups showed no changes of density of enteric neurons in both submucosal and myenteric nervous plexuses in DP of the colon and of enteric glial cells. However, compared to controls and dogs with secondary megacolon, there was a significant decrease in the density of NFP-ir nerve fibers in the longitudinal muscle layer in dogs with idiopathic acquired megacolon. In addition, dogs with idiopathic megacolon display decreased VIP-ir in the myenteric plexus and lamina propria mucosae, and absence of VIP-ir neurons in the submucosal plexus of DP of the colon. Similar alterations, although of lesser severity, may be found in dogs with secondary aquired megacolon. We consider that both idiopathic and secondary aquired megacolon might occur on the basis of a dysplastic changes of VIP-ir enteric neurons.Poznato je da se magakolon javlja kod mesojeda, uključujući mačke i pse, pri čemu je ovo oboljenje daleko učestalije kod mačaka. Na osnovu dosadašnjih saznanja, idiopatski megakolon je čest uzročnik konstipacije kod mačaka i 62% mačaka sa konstipacijom ima idiopatski megakolon. Istovremeno, podaci o psima sa idiopatskim megakolonom veoma su oskudni. U ovom radu je proučavan enterični nervni sistem u dilatiranom delu kolona kod 7 pasa sa idiopatskim megakolonom i 21 psa sa sekundarnim stečenim megakolonom, a rezultati su upoređeni sa normalnim kolonom kod 3 kontrolne zdrave životinje. Tkivni preseci kolona bojeni su klasičnim histološkim i imunohistohemijskim metodama, pri čemu su primenjeni pan-neuronski markeri (NSE, sinaptofizin i neurofilament) i VIP, kao i S-100 protein za detekciju glijalnih ćelija u enteričnim ganglijama. Nisu otkrivene razlike u gustini enteričnih neurona u submukoznom i mijenteričnom pleksusu kod životinja sa megakolonom, kao ni razlike u gustini glijalnih ćelija enteričnih ganglija, u odnosu na kontrolnu grupu životinja. Međutim, u odnosu na kontrolnu grupu, kod životinja sa idiopatskim megakolonom dokazana je smanjena VIP-imunoreaktivnost (ir) u mienteričnom pleksusu i krznu mukoze, kao i kompletno odsustvo VIP-ir neurona u submukoznom pleksusu dilatiranog dela kolona. Slične promene, ali u manjem stepenu, postojale su kod pasa sa sekundarnim stečenim megakolonom. Može da se zaključi da u patogenezi idiopatskog i sekundarnog stečenog megakolona značajnu ulogu imaju displastične promene u VIP-ergičkim neuronima enteričkog nervnog sistema

Karakterizacija matičnih ćelija izolovanih iz zubne pulpe mlečnih zuba dece

Background/Aim. The last decade has been profoundly marked by persistent attempts to use ex vivo expanded and manipulated mesenchymal stem cells (MSCs), as a tool in different types of regenerative therapy. In the present study we described immunophenotype and the proliferative and differentiation potential of cells isolated from pulp remnants of exfoliated deciduous teeth in the final phase of root resorption. Methods. The initial adherent cell population from five donors was obtained by the outgrowth method. Colony forming unit-fibroblast (CFU-F) assay was performed in passage one. Cell expansion was performed until passage three and all tests were done until passage eight. Cells were labeled for early mesenchymal stem cells markers and analysis have been done using flow cytometry. The proliferative potential was assessed by cell counting in defined time points and population doubling time was calculated. Commercial media were used to induce osteoblastic, chondrogenic and adipogenic differentiation. Cytology and histology methods were used for analysis of differentiated cell morphology and extracellular matrix characteristics. Results. According to immunophenotype analyses all undifferentiated cells were positive for the mesenchymal stem cell markers: CD29 and CD73. Some cells expressed CD146 and CD106. The hematopoietic cell marker, CD34, was not detected. In passage one, incidence of CFU-F was 4.7 ± 0.5/100. Population doubling time did not change significantly during cell subcultivation and was in average 25 h. After induction of differentiation, the multicolony derived cell population had a tri-lineage differentiation potential, since mineralized matrix, cartilage-like tissue and adipocytes were successfully formed after three weeks of incubation. Conclusion. Altogether, these data suggest that remnants of deciduous teeth dental pulp contained cell populations with mesenchymal stem cell-like features, with a high proliferation and tri- lineage differentiation potential and that these cultures are suitable for further in vitro evaluation of cell based therapies.Uvod/Cilj. Prošla dekada je bila posebno obeležena naporima na polju korišćenja ex vivo razvijenih i usmeravanih mezenhimskih matičnih ćelija (MSCs), kao sredstva za različite tipove regenerativne terapije. Cilj ove studije bio je da se utvrdi imunofenotip i potencijal za proliferaciju i diferencijaciju ćelija izolovanih iz zubne pulpe mlečnih zuba dece eksfoliranih u periodu kada je koren zuba bio u poslednjoj fazi resorpcije. Metode. Primarna adherentna populacija ćelija poreklom od pet donora dobijena je metodom eksplanta. Prisustvo progenitorskih ćelija koje obrazuju kolonije fibroblasta (CFU-F) pokazano je u prvoj pasaži. Do treće pasaže ćelije su ekspandirane, a potom korišćene za analiziranje. Imunofenotip je određen korišćenjem protočne citometrije. Proliferativni potencijal i vreme udvajanja ćelija (PDT) u kulturi je definisano na osnovu apsolutnog broja ćelija na početku i na kraju svake pasaže. Posle tronedeljne kultivacije ćelija u komercijalnim medijumima za stimulaciju osteogeneze, hondrogeneze i adipogeneze, citološkim i histološkim metodama je određena morfologija ćelija i karakteristike vanćelijskog matriksa. Rezultati. Antigeni koji karakterišu mezenhimske matične ćelije CD29 i CD73 su bili eksprimirani na svim nediferenciranim ćelijama, dok su antigeni CD146 i CD106 bili eksprimirani na ograničenom broju ćelija. Antigen CD34 (karakterističan za ćelije hematopoetske loze) nije bio eksprimiran. Incidencija CFU-F bila je 4,7 ± 0,5/100 ćelija. PDT se nije menjao tokom osam pasaža i u proseku je iznosio 25 h. Posle tronedeljne stimulacije diferencijacije u kulturama sa adipogenim medijumom došlo je do stvaranja ćelija sa masnim kapljicama, a u kulturama sa osteogenim medijumom došlo je do formiranja vanćelijskog matriksa sa deponovanim kalcijumovim solima. U kulturama sa hondrogenim medijumom došlo je do stvaranja tkiva sličnog hrskavici i vanćelijskog matriksa sa glikozaminoglikanima i kolagenom II. Zaključak. Zubna pulpa mlečnih zuba dece sadrži ćelijsku populaciju koja odgovara mezenhimskim matičnim ćelijama prema svojim karakteristikama, ima visok proliferativni potencijal i potencijal da se diferencira u tri ćelijske linije što je čini pogodnom za dalje in vitro analize i evaluaciju ćelijske terapije

Age-related changes in the content of insulin: Like growth factor-l in rat brain

Although there has been extensive research on the effect of IGF-I on muscles and bone tissue, the effect on brain aging has received little attention. We investigated the IGF-I content in brains of differently aged rats. The IGF-I contents in cerebellar and cerebral cortex were found to be higher in immature rats (4-5 days old) compared to young adult (2.5 months old) and middle-aged (7.5-9 months old) rats. However, the decrease of mean IGF-I in middle-aged rats compared to immature animals was statistically significant only in the cerebellar codex. Our results indicate that IGF-I content decreases through the lifespan and maybe selectively in some brain regions.Vršena su istraživanja insulinu sličnog faktora rasta (IGF-I) na mišićno i koštano tkivo, ali je posvećena mala pažnja efektu na mozak u toku starenja. Mi smo ispitivali sadržaj IGF-I u moždanom tkivu pacova različite starosti. Nađeno je da su IGF-I koncentracije u kori malog mozga kao i velikog mozga mladih pacova (4-5 dana starih) više u poređenju sa sadržajima grupe tek-odraslih pacova starosti 2,5 meseca i grupe nešto starijih odraslih pacova (7,5-9 meseci starih). Međutim, smanjenje koncentracije IGF-I sadržaja samo u kori malog mozga nešto starijih pacova (7,5-9 meseci) bilo je značajno u odnosu na vrednosti u novorođenih (4-5 dana starih pacova). Naši rezultati ukazuju da IGF-I opada tokom života i moguće - selektivno u određenim moždanim regionima.nul

Age-related changes in the content of insulin: Like growth factor-l in rat brain

Although there has been extensive research on the effect of IGF-I on muscles and bone tissue, the effect on brain aging has received little attention. We investigated the IGF-I content in brains of differently aged rats. The IGF-I contents in cerebellar and cerebral cortex were found to be higher in immature rats (4-5 days old) compared to young adult (2.5 months old) and middle-aged (7.5-9 months old) rats. However, the decrease of mean IGF-I in middle-aged rats compared to immature animals was statistically significant only in the cerebellar codex. Our results indicate that IGF-I content decreases through the lifespan and maybe selectively in some brain regions.Vršena su istraživanja insulinu sličnog faktora rasta (IGF-I) na mišićno i koštano tkivo, ali je posvećena mala pažnja efektu na mozak u toku starenja. Mi smo ispitivali sadržaj IGF-I u moždanom tkivu pacova različite starosti. Nađeno je da su IGF-I koncentracije u kori malog mozga kao i velikog mozga mladih pacova (4-5 dana starih) više u poređenju sa sadržajima grupe tek-odraslih pacova starosti 2,5 meseca i grupe nešto starijih odraslih pacova (7,5-9 meseci starih). Međutim, smanjenje koncentracije IGF-I sadržaja samo u kori malog mozga nešto starijih pacova (7,5-9 meseci) bilo je značajno u odnosu na vrednosti u novorođenih (4-5 dana starih pacova). Naši rezultati ukazuju da IGF-I opada tokom života i moguće - selektivno u određenim moždanim regionima.nul

Acute effect of ethanol on IgA immunoreactive cells in the intestine-associated immune system

The purpose of this study was to investigate the acute effect of ethanol on mucosa-associated lymphoid tissue at the level of Peyer's patches and the intestinal lamina propria in female rats and to determine whether this action of ethanol is modulated during the estrous cycle. Adult female rats showing proestrus or diestrus day 1 were treated intraperitoneally (ip) with ethanol (4 g/kg). Untreated and saline-injected rats were used as controls. The animals were sacrificed by decapitation 0.5 h after ethanol administration. Immunoglobulin A (IgA) immunoreactive cells were analyzed by indirect immunohistochemistry using mouse anti-rat IgA and a Dako LSAB+ kit. The number of IgA-immunoreactive cells in Peyer's patches was unaltered by ethanol treatment at both phases of the estrous cycle. However, stereological analysis revealed a significant increase in the number of IgA-immunoreactive cells (p lt 0.01) in the intestinal lamina propria following acute ethanol administration at proestrus and on diestrus day 1. The results indicate that the intestinal lamina propria, the effector site of the mucosal immune system, can be affected by a single dose of ethanol at both phases of the estrous cycle

Blockade of Nitric Oxide Synthesis Modulates Rat Immunoglobulin A

Objective: Nitric oxide (NO) is known as a regulator of inflammation and immunity. The purpose of this study was to investigate the influence of this signal molecule on the rat immunoglobulin A (IgA) system using N omega-nitro-L-arginine-methyl ester (L-NAME), which inhibits the activity of all isoforms of NO synthase. Methods: The experiments were performed on adult female Wistar rats showing diestrus day 1 that were treated with L-NAME (30 or 50 mg/kg, s.c.). Untreated and saline-injected animals were used as controls. The rats were sacrificed 3 h following L-NAME or saline administration. The concentration of IgA in serum and intestinal extracts was determined by a sandwich enzyme-linked immunosorbent assay. The number of IgA-expressing cells per area unit of Peyer's patches and the intestinal lamina propria was evaluated using stereological analysis. Results: The results showed that L-NAME decreased the level of IgA in serum and elevated its concentration in intestinal extracts. Additionally, the increased number of IgA+ cells was found in the intestinal lamina propria in both experimental groups. Conclusion: Obtained findings imply that endogenous NO may modulate the IgA system in the rat. Copyright (C) 2009 S. Karger AG, Base

Malignant melanoma in a brown bear (Ursus arctos)

Programme and Proceeding

Expression of heat shock protein 70 (HSP70) in patients with colorectal adenocarcinoma - immunohistochemistry and Western blot analysis

The role of heat shock protein 70 (HSP70) expression has been investigated in various types of tumors. There are only little and controversial data about its clinical relevance in colorectal carcinoma, one of the most common carcinomas observed in humans. In this study we investigated expression of HSP70 in human colonic carcinoma and possible correlation with clinicopathology. To assess patterns (cytosolic and membrane) of HSP70 expression, the 48 surgically removed colorectal adenocarcinomas and 12 normal colonic and rectal mucosal samples were examined by immunohistochemistry and Western-blot. According to results of immunohistochemistry, expression of cytoplasmic HSP72 was significantly higher in colorectal carcinoma compared with normal and adjacent mucosa (p LT 0.01). In addition, there was significant increase in HSP72 expression in lymph node-positive compared to node-nevative group (p LT 0.001). Dukes C2 stage of colonic cancer showed significantly higher immunohistochemical score than Dukes B2 and B1 stage groups (p LT 0.05 i.e. p LT 0.02). There was no relation between expression of HSP72 and degree of tumor differentiation. Using Western blot analyses, we noticed elevated levels of cytosolic HSP70 in colorectal cancer cells compared to normal. Densitometric analysis of blots of plasma membrane HSP70 expression has shown decrease in colorectal cancer cells compared to normal mucosa. According to our results, overexpression of HSP72 in malignant tissues of patients with colorectal carcinoma is related to tumor progression, suggesting that these proteins could play an important role not only in tumorigenesis but also in the development of drug resistance. Further research is necessary to clarify the mechanisms responsible for differential HSP70 expression as well as its definitive role in colorectal cancer

Expression of heat shock protein 70 (HSP70) in patients with colorectal adenocarcinoma - immunohistochemistry and Western blot analysis

The role of heat shock protein 70 (HSP70) expression has been investigated in various types of tumors. There are only little and controversial data about its clinical relevance in colorectal carcinoma, one of the most common carcinomas observed in humans. In this study we investigated expression of HSP70 in human colonic carcinoma and possible correlation with clinicopathology. To assess patterns (cytosolic and membrane) of HSP70 expression, the 48 surgically removed colorectal adenocarcinomas and 12 normal colonic and rectal mucosal samples were examined by immunohistochemistry and Western-blot. According to results of immunohistochemistry, expression of cytoplasmic HSP72 was significantly higher in colorectal carcinoma compared with normal and adjacent mucosa (p LT 0.01). In addition, there was significant increase in HSP72 expression in lymph node-positive compared to node-nevative group (p LT 0.001). Dukes C2 stage of colonic cancer showed significantly higher immunohistochemical score than Dukes B2 and B1 stage groups (p LT 0.05 i.e. p LT 0.02). There was no relation between expression of HSP72 and degree of tumor differentiation. Using Western blot analyses, we noticed elevated levels of cytosolic HSP70 in colorectal cancer cells compared to normal. Densitometric analysis of blots of plasma membrane HSP70 expression has shown decrease in colorectal cancer cells compared to normal mucosa. According to our results, overexpression of HSP72 in malignant tissues of patients with colorectal carcinoma is related to tumor progression, suggesting that these proteins could play an important role not only in tumorigenesis but also in the development of drug resistance. Further research is necessary to clarify the mechanisms responsible for differential HSP70 expression as well as its definitive role in colorectal cancer