Histone deacetylase inhibitor givinostat has ameliorative effect in the colitis model

Purpose: To investigate the effect of givinostat treatment in acetic acid-induced ulcerative colitis model in rats. Methods: Thirty male Wistar albino rats were used. Rats were randomly divided into three equal groups, and colitis was induced on 20 rats by rectal administration of %4 solutions of acetic acid. Twenty rats with colitis were randomly divided into two groups. %0.9 NaCl (saline) solution was administered intraperitoneally to the first group of rats (saline group, n=10) at the dose of 1 mL/kg/day. Givinostat was administered intraperitoneally to the second group rats (Givinostat group, n=10) at the dose of 5 mg/kg/day. Samples were collected for biochemical analysis. Colon was removed for histopathological and biochemical examinations. Results: Plasma tumor necrosis factor-α (TNF-α), pentraxin-3 (PTX-3), and malondialdehyde levels were significantly decreased in the givinostat group compared to the saline group (p<0.05, p<0.001, and p<0.001 respectively; p<0.001, p<0.001, and p<0.001, respectively). Colon TNF-α and prostaglandin F2 alpha (PGF-2) levels were significantly decreased (p<0.05, and p<0.001, respectively). The givinostat group had a significantly lower histologic score than saline group (p<0.001, and p<0.001, respectively). Conclusions: Givinostat, a good protector and regenerator of tissue and an anti-inflammatory agent, may be involved in the treatment of colitis in the future.

Surgery-induced changes in erythrocyte and plasma lipid peroxidation, enzymatic and non-enzymatic antioxidants of female rats: protective role of heparin and pentoxifylline

Background: Intra abdominal adhesions are a complication that occurs after many abdominal surgical procedures, especially gynecological operations. These complications occur by irritating the peritoneum due to such circumstances as infection or surgical trauma, and are considered a pathological part of the healing process of peritoneal injury. It manifests itself with symptoms such as pain, intestinal or urethral obstruction and abdominal abscesses. Oxidative stress due to adhesions plays an important role on adhesion formation. In addition to many researches done at the point of prevention of adhesion and decreasing stress parameters, in this study, it was planned to determine the effect of Heparin (H) and Pentoxifylline (PTX) on malondialdehyde (MDA) and some antioxidant values.Materials, Methods & Results: This study was performed on rats and thirty-seven female Sprague-Dawley rats were randomized into four groups. The first group was sham (Sh) group (n = 7) and laparotomy was performed and 2 mL of 0.9 % NaCl was applied. For all other rats (n = 30) the small intestine was withdrawn and the uterus was uncovered and the anti mesenteric surfaces of the left uterine horn and left abdominal wall were superficially tilted until slight bleeding was seen. Lesion areas have been covered. Two mL 0.9 % NaCl to control (C) group (n = 10), 500 IU heparin to group H, and 25 mg / kg Pentoxifylline to group PTX (n = 10) have been given and then the abdominal incision was closed. The adhesion score of group Sh was found to be more important than C and PTX groups (P < 0.05). The adhesion score of group C was determined to be more significant than group H (P < 0.05). In group Sh, erythrocyte reduced glutathione (GSH) levels were found to be more significant (P < 0.01) than C, H and PTX groups whereas it was found that group C was more significant than group H (P < 0.01).[...]

Surgery-induced changes in erythrocyte and plasma lipid peroxidation, enzymatic and non-enzymatic antioxidants of female rats: protective role of heparin and pentoxifylline

Background: Intra abdominal adhesions are a complication that occurs after many abdominal surgical procedures, especially gynecological operations. These complications occur by irritating the peritoneum due to such circumstances as infection or surgical trauma, and are considered a pathological part of the healing process of peritoneal injury. It manifests itself with symptoms such as pain, intestinal or urethral obstruction and abdominal abscesses. Oxidative stress due to adhesions plays an important role on adhesion formation. In addition to many researches done at the point of prevention of adhesion and decreasing stress parameters, in this study, it was planned to determine the effect of Heparin (H) and Pentoxifylline (PTX) on malondialdehyde (MDA) and some antioxidant values.Materials, Methods & Results: This study was performed on rats and thirty-seven female Sprague-Dawley rats were randomized into four groups. The first group was sham (Sh) group (n = 7) and laparotomy was performed and 2 mL of 0.9 % NaCl was applied. For all other rats (n = 30) the small intestine was withdrawn and the uterus was uncovered and the anti mesenteric surfaces of the left uterine horn and left abdominal wall were superficially tilted until slight bleeding was seen. Lesion areas have been covered. Two mL 0.9 % NaCl to control (C) group (n = 10), 500 IU heparin to group H, and 25 mg / kg Pentoxifylline to group PTX (n = 10) have been given and then the abdominal incision was closed. The adhesion score of group Sh was found to be more important than C and PTX groups (P < 0.05). The adhesion score of group C was determined to be more significant than group H (P < 0.05). In group Sh, erythrocyte reduced glutathione (GSH) levels were found to be more significant (P < 0.01) than C, H and PTX groups whereas it was found that group C was more significant than group H (P < 0.01).[...](AU)

The expression of endothelial and inducible nitric oxide synthase and apoptosis in intestinal ischemia and reperfusion injury under the action of ischemic preconditioning and pentoxifylline

Purpose:To investigate the expression of nitric oxide synthase (NOS) and apoptosis associated with ischemic preconditioning (IPC) and pentoxifylline (PTX) in intestinal ischemia (I) and reperfusion (R) injury.Methods:Thirty male rats were assigned to 5 groups: (CG), no clamping of the superior mesenteric artery (90 minutes); (IR-SS) saline + ischemia (30 minutes) + reperfusion (60 minutes); (IR-PTX) PTX + ischemia (30 minutes) + reperfusion (60 minutes); (IPC-IR-SS) 5 minutes of ischemia + 5 minutes of reperfusion (IPC) + saline + I(30 minutes)+R(60 minutes); and (IPC-IR-PTX) IPC + PTX + I(30 minutes)+ R(60 minutes).Results:The application of IPC and PTX showed a significantly lower immunohistochemistry reaction for active caspase-3 (P<0.05) compared to IR+SS. The number of cells immunoreactive to BCL-2 was higher in the IR-PTX group (P>0.05). The NOS-2 expression (qRTPCR) in the IR-PTX group (P<0.05) was higher than the values for the IPC+IR-SS and IPC-IR-PTX groups. The NOS-3 expression was significantly upper in the IPC-IR-PTX group than in the CG (P<0.05), the IR-SS (P<0.05) and the IR-PTX (P<0.05) groups.Conclusions:The BCL-2 and active caspase-3 showed beneficial effects on PTX and IPC. The expression of NOS-2 and NOS-3 in the IPC and IPC-PTX groups showed no synergistic effect.(AU)

Direct intraperitoneal resuscitation with lidocaine, methylene blue and pentoxiphylline combination does not decreases inflammation after intestinal ischemia-reperfusion injury in rats

PURPOSE:To evaluate the effects of an intraperitoneal solution of methylene blue (MB), lidocaine and pentoxyphylline (PTX) on intestinal ischemic and reperfusion injuryMETHODS:Superior mesenteric artery was isolated and clamped in 36 adult male Sprague Dawley rats. After 60 minutes, clamp was removed and a group received intraperitoneally UNITO solution (PTX 25mg/kg + lidocaine 5mg/kg + MB 2mg/kg), while the other group was treated with warm 0.9% NaCl solution. Rats were euthanized 45 min after drug administration. Lung and bowel were collected for histological evaluation (using Park's score) and determination of myeloperoxidase (MPO) and malondialdehyde (MDA) levels.RESULTS:Control samples showed lymphoplasmocytic infiltrate and crypt necrosis of villi. MPO and MDA measurements shown no differences between treated and control groups.CONCLUSION:The combination of lidocaine, methylene blue and pentoxyphylline administered intraperitoneally at the studied dose, did not decreased histological lesion scores and biochemical markers levels in intestinal ischemia/reperfusion injury.(AU)

ADIÇÃO DE COLESTEROL LIGADO À CICLODEXTRINA NO PROCESSAMENTO DE ESPERMATOZOIDES EQUINOS CRIOPRESERVADOS E ADIÇÃO DE ESTIMULANTES DA MOTILIDADE NO PÓS DESCONGELAMENTO

Este estudo propôs avaliar o efeito da adição de colesterol ligado à ciclodextrina (CLC) antes do processo de criopreservação e estimulantes de motilidade após o descongelamento (Fert Talp - FT e Pentoxifilina - PTX), sobre a cinética espermática (MT, MP, RAP, VCL, VSL, VAP e LIN), integridade de membrana plasmática, produção de peróxido de hidrogênio intracelular, potencial mitocondrial, desestabilização da membrana plasmática e fertilidade in vivo. Foram criopreservados dois ejaculados de 12 garanhões e cada ejaculado foi subdividido em quatro grupos: G1 (sem CLC), G2 (1mg de CLC), G3 (1,5mg de CLC) e G4 (2mg de CLC). No experimento I o objetivo foi comparar os parâmetros espermáticos após a inclusão de CLC (G1-G4) e avaliar a longevidade celular pelo teste de Termorresitência - TTR após o descongelamento: imediatamente (T0), 40 minutos (T40), 80 minutos (T80) e 120 minutos (T120). Além disso, objetivou-se avaliar o potencial mitocondrial e a desestabilização da membrana. No experimento II o objetivo foi comparar os parâmetros espermáticos nos grupos anteriores (G1-G4), com e sem adição do estimulante FT (20% - v/v) após o descongelamento, quanto a longevidade celular (TTR - T0, T40, T80 e T120 minutos) e avaliar a produção de peróxido de hidrogênio intracelular. No experimento III o objetivo foi comparar os parâmetros espermáticos nos grupos anteriores (G1-G4), com e sem adição do estimulante PTX (20% - v/v) após o descongelamento, avaliar a longevidade celular (TTR - T0, T40, T80 e T120 minutos), avaliar a produção de peróxido de hidrogênio intracelular, o potencial mitocondrial e a desestabilização da membrana. Além disso, objetivou-se avaliar a fertilidade do sêmen criopreservado em três grupos: 1) Grupo controle (sem CLC); 2) Grupo 1,5mg de CLC; e 3) Grupo 1,5 mg de CLC + PTX. No experimento I, os grupos G3 e G4 apresentaram valores superiores de VCL, VSL e VAP. No T0 os parâmetros de MT e MP foram superiores no G3. Quanto ao potencial mitocondrial e a desestabilização da membrana não foram observadas diferenças significativas entre os grupos (p<0,05). No experimento II, a adição de FT foi benéfica à velocidade espermática (VCL, VSL e VAP) em alguns grupos experimentais e em tempos diferentes. O G2 com FT demonstrou maior percentual de células com membrana plasmática íntegra e sem peróxido de hidrogênio em comparação aos grupos sem adição do estimulante. No experimento III, a adição de PTX não foi benéfica aos parâmetros de cinética após o descongelamento ao decorrer do tempo em alguns grupos experimentais. No grupo G1 com PTX notou-se superioridade de células com membrana lesionada com peróxido de hidrogênio em comparação ao G4 com PTX e uma maior população celular com membranas íntegras sem peróxido de hidrogênio no grupo G4 com PTX em comparação ao G2 com PTX. A taxa de concepção nos grupos G1, G2 e G3 foram 50%, 66,7% e 66,7%, respectivamente. A adição de CLC melhorou os parâmetros cinéticos in vitro, sendo que as concentrações de 1,5 mg e 2 mg conferiram melhor resistência ao processo de criopreservação. A adição de FT conferiu longevidade e superioridade aos parâmetros de velocidade espermática. Em contrapartida, a adição do estimulante PTX, afetou a longevidade celular, demonstrando efeito prejudicial ao movimento espermático. Além disso, a inclusão de CLC não demostrou prejudicar a taxa de concepção em inseminações realizadas após a ovulação. Mediante os resultados encontrados no atual trabalho, pode-se concluir que as concentrações de CLC que melhor conferiram resistência aos espermatozoides ao processo de criopreservação com consequente incremento cinético às células espermáticas foram 1,5 mg e 2 mg. Os achados com a adição do estimulante FT, considerando-o estimulante de motilidade através do aumento no metabolismo celular, conferiu longevidade e superioridade aos parâmetros de velocidade espermática, sendo essa uma atribuição satisfatória além de conferir também maiores populações celulares com membrana plasmática íntegra sem a produção de peróxido de hidrogênio. Em contrapartida a adição do estimulante PTX, afetou a longevidade celular, demonstrando efeito prejudicial ao movimento espermático. E quanto a fertilidade do sêmen criopreservado, os resultados encontrados com a utilização de colesterol ligado a ciclodextrina não interferiu na fertilidade. Dessa forma, o colesterol ligado à ciclodextrina é benéfico à célula espermática ao processo de criopreservação, entretanto a adição de estimulante causou efeitos divergentes, aumentando os parâmetros e a longevidade celular (FT) ou diminuindo os parâmetros de cinética com o tempo (PTX).

The current study proposed to evaluate the effect of the addition of cholesterol bound to cyclodextrin (CLC) before the cryopreservation process and motility stimulants post thawed (fert talp FT and pentoxifylline PTX) on the spermatic kinetics (MT, MP, RAP, VCL, VSL, VAP and LIN), membrane integrity, production of intracellular hydrogen peroxide, mitochondrial potential, destabilization of the plasma membrane and in vivo fertility. Two ejaculates of 12 stallions were cryopreserved; subdivided into four experimental groups: G1 (no CLC); G2 (1 mg of CLC), G3 (1.5 mg of CLC) and G4 (2 mg of CLC). In experiment I, the objective was to compare spermatic parameters after the inclusion of CLC (G1-G4), to evaluate cell longevity through the thermoresistance test - TTR post thawed: immediately (T0), 40 minutes (T40), 80 minutes (T80) and 120 minutes (T120). In addition, mitochondrial potential and membrane destabilization were evaluated. In experiment II the objective was to compare spermatic parameters after the inclusion of CLC (G1-G4) and the addition of the stimulant FT, to evaluate cell longevity (TTR) post thawed (T0, T40, T80 and T120) and to evaluate the production of intracellular hydrogen peroxide. In experiment II the objective was to compare spermatic parameters in the groups (G1-G4), with and without the addition of the stimulant FT (20% - v/v) post thawed, as to the cellular longevity (TTR T0, T40, T80 and T120 minutes) and to evaluate the production of intracellular hydrogen peroxide. In experiment III the objective was to compare spermatic parameters in the groups (G1-G4), with and without the addition of the stimulant PTX (20% - v/v) post thawed, to evaluate cellular longevity (TTR - T0, T40, T80 and T120 minutes), to evaluate the production of intracellular hydrogen peroxide, the mitochondrial potential and the membrane destabilization. In addition, to evaluate the fertility of the cryopreserved semen in three groups: 1) Control group: no cholesterol addition; 2) Group cholesterol: 1.5 mg, and 3) Group cholesterol 1.5 mg + PTX. In experiment I, groups G3 and G4 presented superior values of VCL, VSL and VAP. In T0, the parameters of MT and MP were superior in G3. As to mitochondrial potential and membrane destabilization, no meaningful differences were observed between the groups (p<0.05). In experiment II, the addition of FT was beneficial to spermatic speed (VCL, VSL and VAP) in some experimental groups and in different times. G2 with FT has demonstrated higher percentage of cells with intact membrane and without hydrogen peroxide in comparison to groups without the addition of the stimulant. In experiment III, the addition of PTX was not beneficial to the kinetics parameters post thawed along time in some experimental groups. In the group G1 with PTX it was observed superiority in cells with injured membrane and with hydrogen peroxide in comparison to G4 with PTX and a higher cell population with intact membranes and without hydrogen peroxide in the group G4 with PTX in comparison to G2 with PTX. The conception rate in the groups G1, G2 and G3 was, respectively, 50%, 66.7% and 66.7%. In this way, it is possible to conclude that the addition of the cholesterol bounded to cyclodextrin is beneficial to the spermatic cell in the cryopreservation process, however, the addition of stimulants post thawed has caused divergent effect. The stimulant FT has provided longevity and superiority to the spermatic velocity parameters. In contrast, the addition of the stimulant PTX has compromised the cellular longevity. In addition, the inclusion of CLC has not affected the conception rate in inseminations carried out after the ovulation.

Pentoxifylline associated to hypertonic saline solution attenuates inflammatory process and apoptosis after intestinal ischemia/reperfusion in rats

PURPOSE:To evaluate intestinal inflammatory and apoptotic processes after intestinal ischemia/reperfusion injury, modulated by pentoxifylline and hypertonic saline.METHODS:It was allocated into four groups (n=6), 24 male Wistar rats (200 to 250g) and submitted to intestinal ischemia for 40 min and reperfusion for 80 min: IR (did not receive any treatment); HS group (Hypertonic Saline, 4ml/kg-IV); PTX group (Pentoxifylline, 30mg/kg-IV); HS+PTX group (Hypertonic Saline and Pentoxifylline). All animals were heparinized (100U/kg). At the end of reperfusion, ileal fragments were removed and stained on hematoxylin-eosin and histochemical studies for COX-2, Bcl-2 and cleaved caspase-3.RESULTS:The values of sO2 were higher on treated groups at 40 minutes of reperfusion (p=0.0081) and 80 minutes of reperfusion (p=0.0072). Serum lactate values were lower on treated groups after 40 minutes of reperfusion (p=0.0003) and 80 minutes of reperfusion (p=0.0098). Morphologic tissue injuries showed higher grades on IR group versus other groups: HS (p=0.0006), PTX (p=0.0433) and HS+PTX (p=0.0040). The histochemical study showed lesser expression of COX-2 (p=0.0015) and Bcl-2 (p=0.0012) on HS+PTX group. A lower expression of cleaved caspase-3 was demonstrated in PTX (p=0.0090; PTXvsIR).CONCLUSION:The combined use of pentoxifylline and hypertonic saline offers best results on inflammatory and apoptotic inhibitory aspects after intestinal ischemia/reperfusion.(AU)

UTILIZAÇÃO DO PROCESSO BIOTECNOLÓGICO DO OVÁRIO ARTIFICIAL PARA AVALIAR A TOXICIDADE DO VITONOLIDO D E DA QUINOXALINA SOBRE OS FOLÍCULOS PRÉ-ANTRAIS CAPRINOS

Embora o número de novos casos de câncer venha aumentando anualmente, o número de sobreviventes também vem crescendo ao longo dos anos, em função do diagnostico precoce e de tratamentos quimioterápicos mais efetivos. Porém, dentre os grandes efeitos indesejáveis da quimioterapia, destaca-se a resistência a multi-drogas (RMD) e a falha ovariana que leva à infertilidade. Portanto, a presente estudo teve dois principais objetivos: 1) imunolocalizar membros da família de proteínas conhecidas como Transportadores ABC (ABCB1, ABCC2 e ABCG2) no tecido ovariano caprino (Fase 1) e 2) avaliar a toxicidade de dois novos compostos com potencial anticâncer, ou seja, o Vitanolido D - VD (Fase 2) e a Quinoxalina - QX (Fase 3) sobre os folículos pré-antrais (primordiais, primários e secundários) cultivados in vitro no interior do tecido ovariano. Na Fase 1, ovários de cabras foram processados e analisados por imuno-histoquímica, qPCR e Western Blottingg para a identificação do ABCB1, ABCC2 e ABCG2. Nas Fases 2 e 3, fragmentos do córtex ovariano foram cultivadas in vitro por 2 e 6 dias em -MEM+ ou -MEM+ adicionado de paclitaxel (PTX controle negativo) ou em diferentes concentrações de VD (1.5, 3.0 e 6.0 M) ou QX (1.5, 3.0 e 6.0 M), respectivamente. Nessas duas fases os folículos foram analisados antes (controle) e após o cultivo na presença dos compostos, considerando-se morfologia e ativação folicular; proliferação (Ki67) e apoptose (TUNEL) das células granulosa, a expressão proteica do ABCB1 e gênica dos fatores reguladores do ciclo celular ciclinas (A - CCNA, B1 - CCNB1, D1 - CCND1 e E1 - CCNE1) e quinases dependentes de ciclina (1 - CDK1; 2 - CDK2, 4 - CDK4 e 6 - CDK6)]. Os resultados mostraram que os três transportadores ABCs foram positivamente imunomarcadas em todas as categorias foliculares estudadas. No tocante aos compostos, a toxicidade do VD6.0 foi similar ao PTX e superior (P <0,05) às demais concentrações (1.5 e 3.0 M), após 2 e 6 dias. A QX também apresentou alta toxicidade aos folículos pré-antrais, especialmente na mais alta concentração (6.0 M), causando um bournout folicular e apoptose, superiores às concentrações de 1.5 e 3.0 M. Ao contrário do VD, a QX inibiu a expressão do ABCB1 nos folículos pré-antrais. Esse trabalho mostrou pela primeira vez que três membros da família dos ABC transportadores foram identificados no ovário caprino. Apesar dos valiosos resultados obtidos nesse estudo, os dois compostos, isto é, o VD e a QX, necessitam ser melhor investigados, sob o ponto de vista da função ovariana, visando auxiliar na sua utilização como agentes quimioterápicos com menor risco para a fertilidade feminina.

Although the number of new cases of cancer increases annually, the number of survivors has also increased over the years, due to the early diagnosis and more effective chemotherapy treatments. However, among the great undesirable effects of chemotherapy, the multidrug resistance (MDR) and the ovarian failure that lead to infertility. Therefore, the present thesis had two main objectives: 1) to immunolocalize members of the family of proteins known as ABC transporters (ABCB1, ABCC2 and ABCG2), responsible for the phenomenon of RMD in goat ovarian tissue (Phase 1) and 2) to evaluate the toxicity of two new compounds with anticancer potential, the Vitanolide D - VD (Phase 2) and Quinoxaline - QX (Phase 3) on the preantral follicles (primordial, primary and secondary) grown in vitro inside the ovarian tissue. In Phase 1, goat ovaries were processed and analyzed by immunohistochemistry, qPCR and Western Blotting for identification of ABCB1, ABCC2 and ABCG2. In Phases 2 and 3, fragments of the ovarian cortex were cultured in vitro for 2 or 6 days in -MEM+ or -MEM+ supplemented with paclitaxel (PTX - negative control) or at different RV concentrations (1.5, 3.0 and 6.0 M) or QX (1.5, 3.0 and 6.0 M), respectively. In these two phases the follicles were analyzed before (control) and after the culture in the presence of the compounds, considering morphology and follicular activation; proliferation (Ki67) and apoptosis (TUNEL) of the granulosa cells, the protein expression of ABCB1 and the gene of cyclin - cyclin regulatory factors (A - CCNA, B1 - CCNB1, D1 - CCND1 and E1 - CCNE1) and cyclin - dependent kinases 1-CDK1; 2-CDK2, 4-CDK4 and 6-CDK6)]. The results showed that the three ABCs transporters were positively immunolabelled in all follicular categories studied. Regarding the compounds, the toxicity of VD6.0 was similar to PTX and higher (P < 0.05) at the other concentrations (1.5 and 3.0 M) after 2 and 6 days. QX also showed high toxicity to the preantral follicles, especially in the highest concentration (6.0 M), causing a follicular burnout and apoptosis, higher than the concentrations of 1.5 and 3.0 M. Unlike VD, QX inhibited the expression of ABCB1 in preantral follicles. This work showed for the first time that three members of the family of ABC transporters were identified in ovary goat. Despite the valuable results obtained in this study, the two compounds, i.e., RV and QX, need to be better investigated, from the point of view of ovarian function, aiming to aid in their use as chemotherapeutic agents with lower risk for female fertility.

Pretreatment with pentoxifylline attenuates lung injury induced by intestinal ischemia/reperfusion in rats / Pré-tratamento com pentoxifilina atenua a lesão pulmonar induzida por isquemia/reperfusão intestinal em ratos

PURPOSE: To investigate the protective effect of pentoxifylline against the lung injury observed after intestinal ischemia (I) followed by a period of reperfusion (R). METHODS: Twenty-eight male Wistar rats were equally divided into 4 experimental groups and operated under ketamine-xylazine anesthesia. (1) Sham: falsely-operated animals; (2) SS+IR: intestinal ischemia was accomplished by clipping the superior mesenteric artery during 60 minutes, with an administration of a standard volume of saline solution (SS) 5 min before the end of the ischemia period; the clip was then releases or a 120-min period of reperfusion; (3) I+PTX+R: ischemia as above, PTX was administered (25 mg/kg) and the gut reperfused as above; (4) PTX+I+PTX+R: Five minutes before arterial occlusion PTX was administered; the superior mesenteric artery was then clipped for 60 minutes. After 55-min ischemia, an additional dosis of PTX was administered; the clip was removed for reperfusion as above. At the 60th min of reperfusion a third dosis of PTX was administered. RESULTS: PTX markedly attenuated lung injury as manifested by significant decreases (all P<0.001 as compared with the SS+IR group) of pulmonary wet/dry tissue weight ratio, total protein content, myeloperoxidase activity and tumor necrosis factor-alpha. Moreover, it was apparent that in the group PTX+I+PTX+R the improvements have been even more significant. CONCLUSION: PTX exerted a protective effect on the lung from the injuries caused by intestinal ischemia/reperfusion.(AU)

OBJETIVO: Avaliar os efeitos protetores da pentoxifilina (PTX) na lesão pulmonar observada após isquemia (I) seguida de reperfusão (R) intestinal. MÉTODOS: Vinte e oito ratos machos foram divididos aleatoriamente em quatro grupos experimentais e operados sobre anestesia quetamina-xilazina. (1) Sham: animais falsamente operados; (2) SS+IR: isquemia intestinal realizada pelo clampeamento da artéria mesentérica superior durante 60 minutos, com a administração de solução salina (SS) 5 minutos antes do período de isquemia, após a retirada do clamp houve a reperfusão por mais 120 minutos; (3) I+PTX+R: isquemia como mencionado anteriormente seguida da administração de PTX (25 mg/Kg) 5 minutos antes do final da isquemia (60 minutos) seguida de reperfusão por mais 120 minutos; (4) PTX+I+PTX+R: 5 minutos antes da isquemia foi administrado PTX, após 55 minutos de isquemia foi administrado outra dose de PTX e a reperfusão mantida por mais 120 minutos, sendo que aos 60 minutos da reperfusão outra dose de PTX foi administrada. RESULTADOS: A pentoxifilina reduziu os marcadores de lesão pulmonar (proteínas totais, malondialdeído, atividade da mieloperoxidase e fator de necrose tumoral) quando comparada com o grupo não tratado (P<0.001), contudo esta redução foi mais significante no grupo PTX+I+PTX+R. CONCLUSÃO: A pentoxifilina exerce efeito protetor no pulmão no trauma causado por isquemia/reperfusão intestinal.(AU)

The effects of pentoxifylline into the kidneys of rats in a model of unilateral hindlimb ischemia/reperfusion injury / Efeitos da pentoxifilina no rim de ratos em um modelo de lesão de isquemia e reperfusão do membro posterior

PURPOSE: To study the role of pentoxifylline (PTX) on remote kidney injury caused by muscle ischemia of left hindlimb of rats. METHODS: After xylazine and ketamine anesthesia, the left hindlimb of rats (n=66) were submitted to 6 hours ischemia (clamping the left common iliac artery). Three groups were used: sham group (SG, n=6), early group (EG, n=30) with reperfusion after 4 hours and late group (LG, n=30) with reperfusion after 24 hours. The saline solution (EG1, n=10 and LG1, n=10) or PTX (40mg.Kg-1) was administered in the reperfusion beginning (EG2, n=10/LG2, n=10) or divided in two doses in the ischemia beginning and reperfusion beginning (EG3, n=10/LG3, n=10). The plasmatic creatinokinase, urea, creatinine, sodium and potassium values were measure and histological samples from left kidney were prepared and H&E stained for scored cellular necrosis and degeneration of kidney tubules and thickness glomerulus determination. The apoptosis index was determined by immunohistochemical expression of the caspase-3. The tests of Mann-Whitney and Kruskal-Wallis (p < 0.05) were applied. RESULTS: The urea (90.5 ± 30.96 mg.dL-1), creatinine (2.28 ± 0.54 mg.dL-1), potassium (16 ± 3.66 mmol.dL-1) and mesangium thickness (0.97 ± 0.42 µm) values were significantly higher in group LG3. There was no significantly difference of caspase 3 expression between EG2 (16.35 ± 1.65 percent) and LG3 (15.57 ± 2.54 percent), and both were significantly worse than SG (9.8 ± 1.98 percent). CONCLUSIONS: The PTX has some protecting effect on remote kidney injury due to hindlimb ischemia/reperfusion injury only in the early phase of reperfusion.(AU)

OBJETIVO: Estudar o papel da pentoxifilina (PTX) nas lesões à distância no rim causadas pela isquemia no membro posterior esquerdo de ratos. MÉTODOS: Sob anestesia com xilazina e quetamina, o membro posterior de ratos (n=66) foi submetido a 6 horas de isquemia pelo clampeamento da artéria ilíaca comum esquerda. Foram estudados três grupos: grupo simulado (SG, n=6), grupo precoce (EG, n=30) após quatro horas de reperfusão e grupo tardio (LG, n=30) após 24 de reperfusão. A solução salina (EG1, n=10 e LG1, n=10) ou a PTX (40mg.Kg-1) foram administradas no início da reperfusão(EG2, n=10/LG2, n=10) ou divididas em duas aplicações no início da isquemia e no início da reperfusão (EG3, n=10/LG3, n=10). Foram medidos os valores plasmáticos da creatinofosfoquinase, uréia, creatinina, sódio e potássio. Amostras do rim esquerdo foram preparadas e coradas em HE para realizar o escore de necrose de células tubulares renais ou presença de obstrução tubular renal na área do córtex renal e da presença do espessamento do mesângio glomerular. O índice de apoptose foi determinado pela expressão imunoistoquímica da caspase-3. Foram aplicados os testes de Mann-Whitney e Kruskal-Wallis (p < 0.05). RESULTADOS: a dosagem de uréia (90,5 ± 30,96 mg.dL-1), creatinina (2,28 ± 0,54 mg.dL-1), potássio (16 ± 3,66 mmol.dL-1) e a espessura do mesângio(0,97 ± 0,42 µm) foram significantemente maiores nos animais do grupo LG3. Não houve diferença significante na expressão da caspase-3 entre os grupos EG2 (16,35 ± 1,65 por cento) e LG3 (15,57 ± 2,54 por cento) e ambos foram significantemente piores que o grupo SG (9,8 ± 1,98 por cento). CONCLUSÃO: A PTX oferece algum efeito protetor nas lesões à distância nos rins de animais submetidos à lesão de isquemia e reperfusão de membro posterior, no período de até quatro horas após a reperfusão.(AU)

Efeito do meio e da dose inseminação sobre a congelabilidade e fertilidade de espermatozóides recuperados da cauda do epidídimo de garanhões

A colheita de espermatozóides da cauda do epidídimo e sua criopreservação representam um grande avanço biotecnológico na equinocultura, e pode ser a última chance de preservação do material genético de garanhões após sua morte inesperada ou lesões que impossibilitem a cobertura ou colheita de sêmen. Os objetivos dessa pesquisa foram comparar a efetividade de dois diluidores utilizados para a recuperação dos espermatozóides epididimários em relação aos índices de congelabilidade determinados laboratorialmente (Experimento I) e as taxas de concepção proporcionadas por cada metodologia ao utilizar baixa dose inseminante (Experimento II). No Experimento I foram utilizados 58 testículos, o ducto epididimário de um testículo de cada garanhão foi lavado com o diluidor sem pentoxifilina?Botu-Sêmen®1 (Controle) e o ducto epididimário do testículo contralateral, com o diluidor contendo 7,18 mM de pentoxifilina?Botu-Turbo®2 (PTX). No Experimento II, foi utilizado um ?pool de espermatozóides? dos testículos de dois garanhões. As amostras foram congeladas e direcionadas às inseminações artificiais em diferentes doses inseminantes, formando os seguintes grupos: 800x106 espermatozóides recuperados com o diluidor sem pentoxifilina (800 Controle), 100x106 espermatozóides recuperados com diluidor sem pentoxifilina (100 Controle) e 100x106 espermatozóides recuperados com o diluidor com pentoxifilina (100 PTX). O índice de concepção para as éguas inseminadas nos grupos 800 Controle, 100 Controle e 100 PTX foi de 68% (11/16), 31,25% (5/16) e 50% (8/16), respectivamente. Frente aos resultados obtidos, conclui-se inseminações artificiais com apenas 100x106 espermatozóides colhidos com o meio diluidor contendo pentoxifilina garantem índices aceitáveis de concepção e maximiza o uso de espermatozóides epididimários congelados

The recovery of spermatozoa from epididymal cauda and its cryopreservation represents a great technological advance, since it is the last possibility to preserve genetic material from dead or deceased valuable males. The aims of this study were to compare the effectiveness of two extenders used for recovery of epididymal sperm in relation to its freezability (Experiment I) and conception rates provided by each methodology using a low insemination dose (Experiment II). In Experiment I, 58 testis were used, epididymal cauda of each stallion were separated and flushed with a skim milk-extender either without (Botu-Sêmen®; Control Group) or with 7,18 mM of pentoxifylline (Botu-Turbo®; PTX Group) and then frozen. In Experiment II, a ?pool? of epididymal sperm from 2 stallions was used. The samples were frozen and artificial insemination was performed using different insemination doses, comprising the following groups: 800x106 sperm recovered without PTX (800 Control); 100x106 sperm recovered without PTX (100 Control) and 100x106 sperm recovered with PTX (100 PTX). The conception rates were 68% (11/16), 31.25% (5/16) e 50% (8/16) for 800 Control, 100 Control and 100 PTX group, respectively. Based in these results, it was possible to conclude that artificial inseminations with 100x106 sperm recovered with PTX guarantee acceptable conception rates and maximize the use of frozen equine epididymal sperm

Correlação da cicloxigenase-2 e p53 com gradação histopatológica dos mastocitomas cutâneos caninos / Cyclooxygenase-2 and p53 correlation with histopathological grade in cutaneous mast cell tumors

O mastocitoma cutâneo canino é uma neoplasia observada na clínica de animais de companhia de grande importância devido a sua alta freqüência, comportamento clínico agressivo e dificuldade de excisão cirúrgica com margens seguras. De todos os métodos de classificação de masteocitomas e estabelecimento do prognóstico, a gradação histopatológica parece ser a de maior consistência preditiva. No entanto, o comportamento clínico de mastocitomas continua imprevisível e técnicas que auxiliem na determinação do prognóstico são necessárias. Foi realizado estudo retrospectivo baseado nos livros de registro do serviço de Patologia Veterinária da faculdade de Medicina Veterinária e Zootencia UNESP, Botucatu, para seleção dos casos com diagnóstico de mastocitoma cutâneo canino, que foram graduados na histopatologia e submetidos a técnica imunoistoquímica para os anticorpos primários anti-p53 mutante e anti-COX2. Não foram observadas diferenças estatísticas entre os valores desses biomarcadores e a gradação histopatológicas, porém todos os graus de mastocitoma apresentaram elevados níveis de COX-2 e p-53 (maiores que 50%) o que corrobora com o comportamento biológico potencialmente maligno dessa neoplasia(AU)

Mast cell tumor is a frequent neoplasia in small animal clinic, and it is very important due its high frequency, aggressive clinical behavior and difficult surgical excision with safe margins. Of all grading methods and prognostic establishment, the histopathological grading is the most confident for predicting the prognosis. Furthermore, the mast cell tumors present an unpredictable clinical behavior and new prognostic tools should be used to identify patients with higher risk of metastasis and death. A restropective study was made using the Pathology Service (from FMVZ, Unesp, Botucatu, SP) register book to select the mast cell tumors, that were graduated eccording to Patnaik et al. (2) and immunohistochemistry technique was applied for p-53 and COX-2 primary antibodies. No statistical differences were seen in both antibodies used and the hispothological grade, but all tumors had more than 50% of the neoplastic cells positive for each antibody, whinch in agreement with the potential malignant behavior of this neoplasia(AU)

Correlação da cicloxigenase-2 e p53 com gradação histopatológica dos mastocitomas cutâneos caninos / Cyclooxygenase-2 and p53 correlation with histopathological grade in cutaneous mast cell tumors

O mastocitoma cutâneo canino é uma neoplasia observada na clínica de animais de companhia de grande importância devido a sua alta freqüência, comportamento clínico agressivo e dificuldade de excisão cirúrgica com margens seguras. De todos os métodos de classificação de masteocitomas e estabelecimento do prognóstico, a gradação histopatológica parece ser a de maior consistência preditiva. No entanto, o comportamento clínico de mastocitomas continua imprevisível e técnicas que auxiliem na determinação do prognóstico são necessárias. Foi realizado estudo retrospectivo baseado nos livros de registro do serviço de Patologia Veterinária da faculdade de Medicina Veterinária e Zootencia – UNESP, Botucatu, para seleção dos casos com diagnóstico de mastocitoma cutâneo canino, que foram graduados na histopatologia e submetidos a técnica imunoistoquímica para os anticorpos primários anti-p53 mutante e anti-COX2. Não foram observadas diferenças estatísticas entre os valores desses biomarcadores e a gradação histopatológicas, porém todos os graus de mastocitoma apresentaram elevados níveis de COX-2 e p-53 (maiores que 50%) o que corrobora com o comportamento biológico potencialmente maligno dessa neoplasia

Mast cell tumor is a frequent neoplasia in small animal clinic, and it is very important due its high frequency, aggressive clinical behavior and difficult surgical excision with safe margins. Of all grading methods and prognostic establishment, the histopathological grading is the most confident for predicting the prognosis. Furthermore, the mast cell tumors present an unpredictable clinical behavior and new prognostic tools should be used to identify patients with higher risk of metastasis and death. A restropective study was made using the Pathology Service (from FMVZ, Unesp, Botucatu, SP) register book to select the mast cell tumors, that were graduated eccording to Patnaik et al. (2) and immunohistochemistry technique was applied for p-53 and COX-2 primary antibodies. No statistical differences were seen in both antibodies used and the hispothological grade, but all tumors had more than 50% of the neoplastic cells positive for each antibody, whinch in agreement with the potential malignant behavior of this neoplasia

Efeitos da pentoxifilina e da n-acetilcisteína em lesões causados por isquemia e reperfusão de órgão esplâncnicos em ratos

A oclusão e a reperfusão das artérias esplâncnicas provoca alterações locais e sistêmicas derivadas principalmente da liberação de substâncias citotóxicas e da interação entre neutrófilos e células endoteliais. Buscando-se estudar os efeitos da pentoxifilina e da n-acetilcisteína em lesões provocadas em um modelo de isquemia e reperfusão (I/R), 60 ratos foram divididos em 6 grupos: SAL/CONT (salina/controle): animais submetidos à cirurgia, mas não à I/R, tratados com solução fisiológica; SAL/ISQ (salina/isquemia): animais submetidos à oclusão da artéria celíaca (AC), artéria mesentérica cranial (AMCr) e artéria mesentérica caudal (AMCa) durante 30 minutos, seguidos de 120 minutos de reperfusão, tratados com solução fisiológica; PTX/CONT: animais submetidos à cirurgia, mas não à I/R, tratados com pentoxifilina; PTX/ISQ: animais submetidos à I/R, tratados com pentoxifilina; NAC/CONT: animais submetidos à cirurgia, mas não à I/R, tratados com n-acetilcisteína; NAC/ISQ: animais submetidos à I/R, tratados com n-acetilcisteína. Os parâmetros avaliados foram pressão arterial média (PAM) carótida, pressão venosa jugular, fluxo sangüíneo na aorta e na AMCr, temperatura esofágica, hematócrito, hemogasometria, histologia intestinal, dosagem de espécies reativas ao ácido tiobarbitúrico (TBARS) em duodeno, jejuno e íleo e de malondialdeído (MDA) plasmático e do íleo. Nos grupos submetidos à I/R, a PAM teve queda significativa após 120 minutos de reperfusão; a pressão venosa não mostrou alterações no decorrer do experimento; houve queda do fluxo sangüíneo na aorta após os 30 minutos de isquemia, com queda mais acentuada ao final do período de reperfusão; o fluxo sangüíneo na AMCr diminuiu significativamente após 120 minutos de reperfusão, porém, em menor grau no grupo NAC/ISQ; a freqüência...

Splanchnic artery occlusion and reperfusion result in local and remote tissue destruction caused by toxic factors released into the circulation and by neutrophil-endothelial cell interactions. In order to evaluate the effects of pentoxifylline (PTX) and n-acetilcysteine (NAC) on ischemia/reperfusion model (I/R), sixty rats were allocated into six groups: 1) SAL/CONT: Sham operation + saline; 2) SALI/ISQ: 30 min celiac, cranial and caudal mesenteric arteries occlusion + 120 min reperfusion + saline; 3) PTX/CONT: Sham operation + PTX (50 mg/kg); 4) PTX/ISQ: I/R + PTX; 5) NAC/SAL: Sham operation + NAC (430 mg/kg) and 6) NAC/ISQ: I/R + NAC. In all groups above mean arterial blood pressure (MABP), vein pressure, aorta and cranial mesenteric artery blood flow, heart rate, esophagus temperature, hematocrit, blood gas determination, histology, thiobarbituric acid reaction species (TBARS) and malondialdehyde (MDA) levels were determined. In I/R groups, MABP decreased significantly 120 minutes after reperfusion (p<0,05); as no difference in vein pressure were observed. Aorta blood flow decreased 30 minutes after ischemia, decreasing dramatically following restoration of blood flow. Cranial mesenteric artery blood flow decreased significantly after reperfusion, although the fall in NAC/ISQ group was attenuated. During the assays, heart rate was reduced, temperature decreased progressively and metabolic acidosis took place. Pentoxiffyline prevented histopathologic signs of injury in duodenum, jejunum and ileum, as well as n-acetilcysteine prevented ileum histopathologic signs of injury. I/R caused TBARS increase in NAC group jejunum and saline group ileum, but no difference was observed in plasmatic and ileal MDA. The present study allowed concluding that PTX was more efficient in preventing histophatologic injury than NAC, but neither PTX nor NAC...