A imunomarcação positiva para c-kit está associada com a presença de células análogas às intersticiais de Cajal no músculo ciliar?

PURPOSE: Interstitial cells of Cajal were identified in the gastrointestinal tract of several species, with close relation to the enteric nervous system. Since it was recognized that interstitial cells of Cajal express the gene product of c-kit, we performed immunohistochemistry for c-kit protein in ciliary muscle specimens of monkeys' eyes. METHODS: Eight eyes from four adult male new world monkeys (Cebus apella) were studied. After blocking endogenous peroxidase activity and nonspecific protein binding, 1:100 dilution of mouse monoclonal antibody against c-kit human oncoprotein was applied to tissues. Antigen-antibody reaction was visualized using the avidin-biotinylated horseradish peroxidase complex in each slide. RESULTS: We observed some groups of fusiform c-kit expressing cells located amongst muscle bundles of the ciliary muscle. Other pigment cells and mast cells were also observed. CONCLUSION: C-kit expressing cells observed in the ciliary muscle of Cebus apella, showed no similarity to melanocytes or mast cells and they could be associated with their gastrointestinal interstitial cells of Cajal counterpart.OBJETIVO: As células intersticiais de Cajal estão presentes no trato gastrintestinal de diversas espécies animais, em íntima relação com o sistema nervoso entérico. Uma vez que as células intersticiais de Cajal expressam o produto do gene c-kit, realizou-se um ensaio imuno-histoquímico a fim de se verificar a marcação da proteína c-kit no músculo ciliar de amostras de olhos de macacos. MÉTODOS: Oito olhos de quatro macacos do novo mundo (Cebus apella) foram estudados. Após bloqueio da peroxidase endógena e de ligação protéica não específica, os tecidos receberam aplicação de anticorpos de camundongos antioncoproteína c-kit humana (1:100). A reação antígeno-anticorpo foi verificada através da aplicação do complexo avidina-biotinilada-peroxidase em cada lâmina. RESULTADOS: Foram observados grupos de células que expressam c-kit, localizadas entre as fibras do músculo ciliar. Mastócitos e outras células pigmentadas também foram observadas. CONCLUSÃO: Algumas células que expressam c-kit, observadas no músculo ciliar de Cebus apella, não mostraram similaridade com mastócitos ou melanócitos e podem ser classificadas como análogas das células intersticiais de Cajal gastrintestinais.Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)FAPES

Treatment of canine multicentric lymphoma through vascular access port vs. peripheral venous catheter

Background: Vascular access port (VAP) was developed for the administration of chemotherapeutic agents, minimizing local drug reactions and complications associated with migration of peripheral venous catheter (PVC) in humans. The device is widely used in human oncology and has gained importance in veterinary oncology, especially in long treatment regimens, as in the case of canine lymphoma. VAP favors therapy and the animals life quality. The aim of this study was to describe the use of VAP in dogs, comparing to PVC access, during canine lymphoma chemotherapeutic treatment. Materials, Methods & Results: Eleven dogs with multicentric lymphoma which required chemotherapy were selected for the study. The dogs were randomly allocated to two groups with five and six animals, and each group received the chemotherapy protocol through the PVC (n= 5) or VAP (n= 6). For the sake of standardization, assessments were made whenever the dogs received vincristine sulfate, despite the use of the infusion system in all sessions of the Madison- Wisconsin protocol. A VAP was implanted into the right external jugular vein of six dogs under inhalational anesthesia, using the Seldinger technique. Systolic blood pressure (SBP) levels and handling time during chemotherapy sessions were compared in both groups in three time periods during the procedures: 10 min after arrival to each chemotherapy (P1); immediately after placement of the PVC or puncture of the VAP reservoir (P2); and at the end of chemotherapy (P3). The arithmetic mean of five consecutive assessments was used in each time period. In the chemotherapy sessions, the mean of SBP variation decreased statistically significant in the VAP group compared to PVC group. SBP decreased from P1 to P2 and from P1 to P3 in all sessions (S1, S2, and S3) in the VAP group, and increased in the PVC group. The handling time of VAP group was 110.6 ± 8.4 s, compared to 219.2 ± 24.7 s (mean ± standard error) in the PVC group, showing statisti¬cally significant difference (P < 0.001). VAP surgical implantation time averaged 37 min, decreasing gradually from the first (55 min) to the last patient (21 min). Discussion: SBP levels suggest that the VAP group was calmer from the beginning to the end of the sessions, showed lower SBP levels, and required shorter handling time than did the PVC group. Blood pressure is one of the most objec¬tive ways to assess welfare or stress in dogs. When dog feels threatened or scared, its body automatically enters a state of emergency and, among several changes, blood pressure increases. VAP surgical implantation in dogs have easy learning, as previously described, proven by implantation time progressive reduction. The Seldinger technique is the method of choice for catheter implantation in humans. Dissection of the jugular vein is an alternative, however, the technique with a single incision and venipuncture is less invasive than its modifications. The jugular vein was used because is the site of choice for central accesses in veterinary practice, with a shorter path to the right atrium and smaller rates of catheter mis¬placement, reducing the risk of pneumothorax, venous thrombosis, and pinch-off syndrome. VAP surgical implantation in dogs have easy learning, proven by the implantation time progressive reduction. The study confirmed that VAP promoted animal welfare, shortened chemotherapy sessions, and caused less discomfort to dogs treated for multicentric lymphoma, as indicated by the reduction in SBP, when compared to the PVC group

Evaluation of the potential antiallergic flavonoids: synergistic influence and liposomal systems

Os problemas decorrentes de doenças alérgicas é tema em destaque atualmente devido ao aumento de pessoas que vêm apresentando sintomas e sofrendo as consequências de mudanças aceleradas nos costumes da nossa sociedade. Apesar das diversas terapias oferecidas, pacientes alérgicos ainda sofrem com efeitos colaterais decorrentes do uso de medicamentos anti-alérgicos. Assim, a busca por novas alternativas que possam amenizar os sintomas alérgicos torna-se relevante. As substâncias naturais tem sido alvo de intensa investigação devido às propriedades farmacológicas no tratamento de diversas doenças incluindo as alérgicas. Neste contexto, este trabalho foi focado no extrato Vimang e seu componente majoritário, a mangiferina (Mgf), que tem apresentado atividades biológicas diversas. Sabendo-se que a atividade biológica de um extrato pode ser potencializada pelo sinergismo entre seus componentes, foi avaliado o efeito sinérgico de substâncias isoladas do Vimang, tais como mangiferina, quercetina, catequina, ácido gálico e benzóico. Esses componentes foram combinados e as misturas avaliadas quanto à capacidade em inibir a desgranulação mastocitária. O Vimang, a quercetina e mangiferina inibem a desgranulação mastócitária enquanto catequina, ácido gálico e benzóico não são ativos; e porisso, não foram incluídos na determinação do isobolograma de aditividade, adotado no estudo de sinergismo. O isobolograma de aditividade mostra sinergismo para as concentrações de mangiferina iguais a 5, 10, 20 e 40 ?M e quercetina iguais a 0,2, 1 e 1,6 ?M; e mostra antagonismo para mangiferina 80 ?M e quercetina 0,4 e 0,8 ?M. Tais resultados sugerem que altas concentrações de mangiferina atrapalham a interação sinérgica entre estas substâncias e altas concentrações de quercetina exercem um efeito contrário, ou seja, favorecem o sinergismo entre quercetina e mangiferina. A baixa solubilidade de flavonóides em meio fisiológico levou ao estudo da interação da Mgf com lipossomos de Dimiristoil-L-?-Fosfatidilcolina (DMPC) para aplicação nos ensaios biológicos. A Interação da Mgf com lipossomos de DMPC, bem como a estabilidade lipossomal, foram avaliadas quanto aos efeitos de pH, força iônica e presença de colesterol. O pH influencia a eficiência de incorporação (EI) da Mgf que é maior em pHs ácidos. Em soluções de pH ácido ou fisiológico há aumento de tamanho do lipossomo na presença de Mgf, o que reforça a hipótese de interação desta com a bicamada lipídica. O aumento da força iônica, na ausência de Mgf, diminui a estabilidade do lipossomo. Na ausência de sal, a Mgf favorece a formação de agregados e diminui a estabilidade dos lipossomos e na presença de sal, a Mgf previne a agregação e estabiliza os agregados lipossomais. A EI da Mgf nos lipossomos de DMPC preparados pelo método da injeção etanólica, em pH fisiológico, foram ao redor de 13%. A tentativa de otimizar a EI foi realizada pela adição de colesterol em diferentes proporções lipídio:colesterol. Um aumento de 13% para 17% foi observado para a proporção DMPC:colesterol (9:1). A Mgf livre inibe a desgranulação dos mastócitos de forma concentração-dependente; mas a presença de lipossomos, preparados pelo método da injeção etanólica não altera significativamente esse perfil. A EI da mangiferina em lipossomos de DMPC:colesterol (9:1), preparados pelo método do filme lipídico, foi igual a 45%. Os resultados de potencial antialérgico, para este caso, mostraram que os lipossomos de DMPC favorecem o potencial antialérgico da Mgf em concentrações acima de 25 ?M. Esses resultados abrem perspectivas na busca de sistemas lipossomais mais estáveis e que possam ser aplicados em ensaios biológicos in vitro e in vivo.The problems due to allergic diseases are currently a highlighted topic due to the increase of people who are showing symptoms and suffering the consequences of rapid changes in the habits of our society. Despite various therapies offered, allergic patients still suffer side effects from the use of anti-allergic drugs. Thus, the search for new alternatives that can alleviate the allergic symptoms becomes relevant. Natural substances have been the subject of intense research due to the pharmacological properties in the treatment of several diseases, including allergic ones. In this context, this work was focused on Vimang extract and its major component, the mangiferin (Mgf), which has shown several biological activities. Given that the biological activity of an extract can be enhanced by the synergism between its components, the synergistic effect of compounds isolated from Vimang such as mangiferin, quercetin, catechin, gallic acid and benzoic acid was evaluated. These components were combined and the mixtures tested for their ability to inhibit mast cell degranulation. The Vimang, quercetin and mangiferin inhibit mast cell degranulation while catechin, gallic acid and benzoic acid are not active and for that reason they were not included in the determination of the isobologram adopted in the synergism study. The isobologram of additivity shows synergism for the concentration of mangiferin equal to 5, 10, 20 and 40 ?M and quercetin equal 0.2, 1 and 1.6 ?M, and shows antagonism for mangiferin 80 ?M and quercetin 0.4 and 0.8 ?M. The low solubility of flavonoids in physiological medium led to the study of the interaction of Mgf with liposomes of L-?-dimyristoyl-phosphatidylcholine (DMPC) for application in biological assays. The interaction of Mgf with DMPC liposomes as well as the liposomal stability was evaluated considering the effects of pH, ionic strength and cholesterol presence. The pH influences the incorporation efficiency (IE) of mangiferin, which is higher in acidic pHs. In solutions of acidic or physiological pH, the liposome size increases in the presence of Mgf, which reinforces the hypothesis of interaction of Mgf with the lipid bilayer. The increased ionic strength in the absence of Mgf decreases the stability of the liposome. In the absence of salt, Mgf favors the formation of aggregates and decreases the stability of the liposomes. When in presence of salt, Mgf prevents the aggregation and stabilizes the liposomal aggregates. The IE of mangiferin in DMPC prepared by the ethanol injection method, at physiological pH, were around 13%. The attempt to optimize the IE was performed by the addition of cholesterol in different proportions lipid:cholesterol. An increase from 13% to 17% was observed for the proportion of DMPC:cholesterol (9:1). Free mangiferin inhibits mast cells degranulation in a dose-dependent manner, but the presence of liposomes prepared by the ethanol injection method does not significantly change this profile. The IE of mangiferin in liposomes of DMPC:cholesterol (9:1), prepared by the method of lipidic film was equal to 45%.The results of antiallergic potential in this case show that DMPC liposomes favor the mangiferin antiallergic potential at concentrations above 25 mM. These results open perspectives in the search for more stable liposomal systems and can be applied to biological assays in vitro and in vivo

Comparison of the properties of compacted and porous lamellar chitosan-xanthan membranes as dressings and scaffolds for the treatment of skin lesions

Compacted and porous lamellar membranes of xanthan (Xn) and chitosan (Ch) at mass ratios of 1 : 1 and 1.2 : 0.8 were prepared and tested to verify possible applications in the treatment of skin lesions. All membranes were prepared by complexation of the polysaccharides in solution and subsequent casting. The porous membranes were obtained by adding either Tween 80 or Pluronic F68 to the polysaccharide complexes before casting. Membranes prepared in the absence of surfactants at a mass ratio of Xn to Ch of 1 : 1 proved ideal for use as wound dressings, as they were thin (around 0.10 mm in thickness) and transparent and had low in vitro cytotoxicity to L929 cells, a tensile strength at break of 25 MPa, water absorption after 24 h of around 86 g/g and simulated body fluid absorption of 16% and adequate stability in the presence of the same solutions. Membranes prepared at the mass ratio of Xn to Ch of 1 : 1 in the presence of Pluronic F68 showed the most favorable characteristics for application as scaffolds for tissue engineering. These membranes consisted of a matrix with interconnected pores which were distributed homogeneously throughout the structure and had a thickness of 1.84 mm, high capacity for FBS uptake (around 18 g/g) and cell culture medium uptake (8.6 g/g), a loss of mass in the culture medium of 33% after 144 h, and low in vitro toxicity to L929 cells. In conclusion, membranes of Ch and Xn produced in the presence or absence of the surfactant Pluronic F68 have a high potential for use as scaffolds in tissue engineering or as dermal dressings, respectively, whereas in contrast, membranes prepared in the presence of Tween 80, regardless of the mass ratio of Xn to Ch, were very cytotoxic to L929 cells and therefore were not appropriate for any of the proposed applications. (C) 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Revisiting Szeptyckitheca Betsch & Weiner (Collembola, Symphypleona, Sminthuridae): new species, updated diagnoses, and a key

Szeptyckitheca Betsch & Weiner is a genus of Sphyrothecinae (Sminthuridae) similar to Sphyrotheca Börner, with 13 nominal species. Most descriptions of Szeptyckitheca taxa lack valuable data in face of the current taxonomy of Symphypleona. In this study the previously described species of the genus were surveyed, aiming to provide updated diagnoses for them. Three species were also transferred to Szeptyckitheca: Sphyrotheca karlarum Palacios-Vargas, Vázquez & Cuéllar, 2003, S. peteri Palacios-Vargas, Vázquez & Cuéllar, 2003, and S. koreana Betsch & Weiner, 2009, based on trochanteral and/or dental chaetotaxy. Two new Brazilian species of the genus are described and illustrated, S. andrzeji Medeiros, Bellini & Weiner, sp. nov., with a remarkable reduced ventral dental chaetotaxy not seen in other Neotropical species, and S. cyanea Oliveira, Medeiros & Bellini, sp. nov. with a distinctive large set of head vertex spines (18). Finally, an updated key to all the valid species of the genus is presented

Efeito do dinitrato de isossorbida sobre a hiperalgesia corneana induzida por lipopolissacarídeo

OBJETIVO: Avaliar o efeito do dinitrato de isossorbida (DNI) sobre a hiperalgesia corneana e a infiltração de neutrófilos em um modelo experimental de ceratite superficial induzida pelo lipopolissacarídeo (LPS). MÉTODOS: A hiperalgesia foi estudada através da indução de uma ceratite inflamatória em ratos (n = 60), pela exposição da córnea ao LPS (LPS +). Após a exposição, os olhos foram tratados durante quatro dias, com as soluções tópicas: DNI (200 mg, 65 mg e 20 mg), prednisolona 1% (PRED) ou Veículo. Olhos controles foram expostos apenas à solução salina (LPS -). Para avaliar a dor ocular, contouse o número de piscadas em 40 segundos, após instilação de uma gota de capsaicina 0,01 mM. A análise histopatológica foi realizada para avaliação da infiltração neutrofílica. RESULTADOS: Foram observadas áreas esbranquiçadas no estroma corneano dos olhos com a ceratite induzida pelo LPS, entre os dias 3 e 15 dias. Esses olhos (LPS +) apresentaram um número significativamente maior de piscadas que os LPS - (P = 0,019) à estimulação química. O tratamento tópico com o DNI reduziu o número de piscadas observadas (P = 0,010). Da mesma forma, olhos expostos ao LPS e tratados com DNI demonstraram redução significativa na infiltração neutrofílica (P = 0,0031). CONCLUSÃO: Baixas doses de DNI reduziram a hiperalgesia corneana e a infiltração neutrofílica nesse modelo de ceratite. Dessa forma, doadores de óxido nítrico, como o DNI, poderão ser úteis no futuro ao tratamento clínico de diversas condições dolorosas da superfície ocular

Treatment of canine multicentric lymphoma through vascular access port vs. peripheral venous catheter

Background: Vascular access port (VAP) was developed for the administration of chemotherapeutic agents, minimizing local drug reactions and complications associated with migration of peripheral venous catheter (PVC) in humans. The device is widely used in human oncology and has gained importance in veterinary oncology, especially in long treatment regimens, as in the case of canine lymphoma. VAP favors therapy and the animals life quality. The aim of this study was to describe the use of VAP in dogs, comparing to PVC access, during canine lymphoma chemotherapeutic treatment. Materials, Methods & Results: Eleven dogs with multicentric lymphoma which required chemotherapy were selected for the study. The dogs were randomly allocated to two groups with five and six animals, and each group received the chemotherapy protocol through the PVC (n= 5) or VAP (n= 6). For the sake of standardization, assessments were made whenever the dogs received vincristine sulfate, despite the use of the infusion system in all sessions of the Madison- Wisconsin protocol. A VAP was implanted into the right external jugular vein of six dogs under inhalational anesthesia, using the Seldinger technique. Systolic blood pressure (SBP) levels and handling time during chemotherapy sessions were compared in both groups in three time periods during the procedures: 10 min after arrival to each chemotherapy (P1); immediately after placement of the PVC or puncture of the VAP reservoir (P2); and at the end of chemotherapy (P3). The arithmetic mean of five consecutive assessments was used in each time period. In the chemotherapy sessions, the mean of SBP variation decreased statistically significant in the VAP group compared to PVC group. SBP decreased from P1 to P2 and from P1 to P3 in all sessions (S1, S2, and S3) in the VAP group, and increased in the PVC group. The handling time of VAP group was 110.6 ± 8.4 s, compared to 219.2 ± 24.7 s (mean ± standard error) in the PVC group, showing statisti¬cally significant difference (P < 0.001). VAP surgical implantation time averaged 37 min, decreasing gradually from the first (55 min) to the last patient (21 min). Discussion: SBP levels suggest that the VAP group was calmer from the beginning to the end of the sessions, showed lower SBP levels, and required shorter handling time than did the PVC group. Blood pressure is one of the most objec¬tive ways to assess welfare or stress in dogs. When dog feels threatened or scared, its body automatically enters a state of emergency and, among several changes, blood pressure increases. VAP surgical implantation in dogs have easy learning, as previously described, proven by implantation time progressive reduction. The Seldinger technique is the method of choice for catheter implantation in humans. Dissection of the jugular vein is an alternative, however, the technique with a single incision and venipuncture is less invasive than its modifications. The jugular vein was used because is the site of choice for central accesses in veterinary practice, with a shorter path to the right atrium and smaller rates of catheter mis¬placement, reducing the risk of pneumothorax, venous thrombosis, and pinch-off syndrome. VAP surgical implantation in dogs have easy learning, proven by the implantation time progressive reduction. The study confirmed that VAP promoted animal welfare, shortened chemotherapy sessions, and caused less discomfort to dogs treated for multicentric lymphoma, as indicated by the reduction in SBP, when compared to the PVC group

Treatment of Canine Multicentric Lymphoma Through Vascular Access Port Versus Peripheral Venous Catheter

Background: Vascular access port (VAP) was developed for the administration of chemotherapeutic agents, minimizing local drug reactions and complications associated with migration of peripheral venous catheter (PVC) in humans. The device is widely used in human oncology and has gained importance in veterinary oncology, especially in long treatment regimens, as in the case of canine lymphoma. VAP favors therapy and the animals’ quality of life. The aim of this study was to describe the use of VAP in dogs, comparing to PVC access, during canine lymphoma chemotherapeutic treatment. Materials, Methods & Results: Eleven dogs with multicentric lymphoma which required chemotherapy were selected for the study. The dogs were randomly allocated to two groups with five and six animals, and each group received the chemotherapy protocol through the PVC (n= 5) or VAP (n= 6). For the sake of standardization, assessments were made whenever the dogs received vincristine sulfate, despite the use of the infusion system in all sessions of the Madison-Wisconsin protocol. A VAP was implanted into the right external jugular vein of six dogs under inhalational anesthesia, using the Seldinger technique. Systolic blood pressure (SBP) levels and handling time during chemotherapy sessions were compared in both groups in three time periods during the procedures: 10 min after arrival to each chemotherapy (P1); immediately after placement of the PVC or puncture of the VAP reservoir (P2); and at the end of chemotherapy (P3). The arithmetic mean of five consecutive assessments was used in each time period. In the chemotherapy sessions, the mean of SBP variation decreased statistically significant in the VAP group compared to PVC group. SBP decreased from P1 to P2 and from P1 to P3 in all sessions (S1, S2, and S3) in the VAP group, and increased in the PVC group. The handling time of VAP group was 110.6 ± 8.4 s, compared to 219.2 ± 24.7 s (mean ± standard error) in the PVC group, showing statistically significant difference (P < 0.001). VAP surgical implantation time averaged 37 min, decreasing gradually from the first (55 min) to the last patient (21 min).Discussion: SBP levels suggest that the VAP group was calmer from the beginning to the end of the sessions, showed lower SBP levels, and required shorter handling time than did the PVC group. Blood pressure is one of the most objective ways to assess welfare or stress in dogs. When dog feels threatened or scared, its body automatically enters a state of emergency and, among several changes, blood pressure increases. VAP surgical implantation in dogs have easy learning, as previously described, proven by implantation time progressive reduction. The Seldinger technique is the method of choice for catheter implantation in humans. Dissection of the jugular vein is an alternative, however, the technique with a single incision and venipuncture is less invasive than its modifications. The jugular vein was used because is the site of choice for central accesses in veterinary practice, with a shorter path to the right atrium and smaller rates of catheter misplacement, reducing the risk of pneumothorax, venous thrombosis, and pinch-off syndrome. VAP surgical implantation in dogs have easy learning, proven by the implantation time progressive reduction. The study confirmed that VAPpromoted animal welfare, shortened chemotherapy sessions, and caused less discomfort to dogs treated for multicentric lymphoma, as indicated by the reduction in SBP, when compared to the PVC group