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A intoxica??o de bovinos por Pteridium (aquilinum) arachnoideum em Santa Catarina e a identifica??o das bact?rias envolvidas nos infartos do quadro agudo / The cattle poisoning by Pteridium (aquilinum) arachnoideum in Santa Catarina and the identification of bacteria involved in infarcts in the acute poisoningJ?NCK, Fernanda 16 December 2014 (has links)
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Previous issue date: 2014-12-16 / CAPES / Pteridium (aquilinum) arachnoideum is a cosmopolitan plant, responsible for heavy losses in cattle for heavy losses in almost all Brazil. This plant causes three clinical pictures: an acute disease, characterized by hemorrhages and fever, and two chronic diseases characterized by bladder tumors and carcinomas of the superior digestive tract. In Santa Catarina, in a retrospective study, the acute poisoning is the form that prevailed. Most of the cases ocurred in the autumn and the main clinical signs observed were fever and faeces with blood. The macroscopic lesions were widespread hemorrhages and infarcts mainly in lung, liver, intestine and lymphnodes, which were characterized by necrosis associated with groups of basophylic bact?ria, sometimes with formation of bubbles of gas. In the bone marrow there was rarefaction or absence of hematopoietic tissue. The experimental reproduction of the disease was realized in four cattle, two vaccinated against clostridioses and two not vaccinated. These cattle received doses of 20, 20, 14 and 10g/kg/day, and they died after 82, 94, 46 and 76 days, when they had ingested 149, 180, 71 and 75% of the plant in relation to their weight. The course of the clinical signs was 5, 4, 1 and 5 days, and at post-mortem examination the lesions found were hemorrhages of varied degrees and locations. Liver infarcts were found in bovines 2, 3 and 4, and in the intestine in all the cattle. The histological lesions were characterized by rarefaction and absence of hematopoietic tissue in the bone marrow, necrosis and bacterial aggregates in the liver, lung, intestine and lymphnodes. Histopathology did not reveal inflammatory reaction and if present the intensity was slight. Samples of organs with infarcts collected at necropsy, in the spontaneous and experimental intoxication, sown in Tarozzi medium, produced gas. Five samples caused death in mice after inoculation of the medium. At necropsy the carcasses had putrid smell, the subcutaneous tissue was red and there was edema and red liquid in the abdominal cavity. Two mice that were sacrificed presented inflammatory reaction in the place of the application, characterized by areas of adherence of the skin to the subcutaneous tissue and presence of abscesso, and four presented putrid smell at necropsy. The impression of of the liver capsula of the mice that died or got sick, revealed small Gram positive rods. Histopathology of the mice that died, revealed in the skeletal musculature of the thighs edema between the fibers with necrosis and eosinofilia of fibers and great amount of small basophylic rods, associated with slight inflammatory mononuclear infiltration and hemorrhage. In the skin also inflammatory filtrate was observed, with edema in the derma and great amount of small basophylics rods. The rest of the mice were sacrificed and no alterations were found. The identification by Chain reaction of Polimerase (PCR) of the liver of the mice that had died, in Tarozzi medium, resulted in Clostridium septicum. / Pteridium (aquilinum) arachnoideum ? planta cosmopolita, respons?vel por perdas vultuosas na cria??o de bovinos em quase todas as regi?es do Brasil. Esta planta ? respons?vel por causar tr?s quadros cl?nicos: um quadro agudo, caracterizado por hemorragias e febre, e dois quadros cr?nicos caracterizados por tumores de bexiga e do trato digest?rio superior. Em Santa Catarina, em estudo retrospectivo, a intoxica??o aguda ? a forma que prevaleceu sobre as demais. A maioria dos casos ocorreu no outono e os principais sinais cl?nicos observados foram febre e fezes com sangue. As les?es macrosc?picas encontradas foram hemorragias generalizadas e infartos principalmente em pulm?o, f?gado, intestino e linfonodo, os quais se caracterizavam por necrose associada a agregados bacterianos bas?filos, em alguns casos com forma??o de bolhas de g?s. Na medula ?ssea havia rarefa??o ou aus?ncia do tecido hematopo?tico. A reprodu??o experimental da doen?a foi realizada em quatro bovinos, dois vacinados contra clostridioses e dois n?o vacinados. Estes receberam doses de 20, 20, 14 e 10g/kg/dia de Pteridium (aquilinum) arachnoideum, e morreram com 82, 94, 46 dias e 76 dias, quando tinham ingerido 149, 180, 71 e 75% de planta em rela??o ao peso vivo. A evolu??o dos sinais cl?nicos foi de 5, 4, 1 e 5 dias, e, ? necropsia, as les?es consistiram de hemorragias em variados graus e localiza??es. Infartos de f?gado foram encontrados nos Bovinos 2, 3 e 4 e no intestino em todos os bovinos. As les?es histol?gicas se caracterizaram por rarefa??o e aus?ncia de tecido hematopo?tico na medula ?ssea, necrose e agregados bacterianos no f?gado, pulm?o, intestino e linfonodo. As les?es histol?gicas n?o revelaram rea??o inflamat?ria e quando presente, a intensidade era leve. Amostras de ?rg?os com infartos coletadas de necropsias nas intoxica??es espont?nea e experimental foram semeadas no meio de cultivo de Tarozzi e produziram g?s. Cinco amostras causaram a morte dos camundongos ap?s inocula??o do meio. ? necropsia desses camundongos verificou-se carca?as com cheiro p?trido, tecido subcut?neo avermelhado e com edema e l?quido avermelhado na cavidade abdominal. Dois camundongos que foram eutanasiados apresentaram rea??o inflamat?ria no local da aplica??o, caracterizada por ?reas de ader?ncia da pele com o tecido subcut?neo e abscessos; quatro exalavam cheiro p?trido na hora da realiza??o da necropsia. A impress?o da c?psula do f?gado dos camundongos que morreram e que ficaram doentes, revelou bastonetes Gram-positivos. ? histologia dos camundongos que morreram verificou-se na musculatura esquel?tica da regi?o da coxa, edema e hemorragia entre as fibras, necrose e eosinofilia de fibras e grande quantidade de bastonetes bas?filos, associado a infiltrado inflamat?rio mononuclear leve e hemorragia. Na pele tamb?m verificou-se, infiltrado inflamat?rio, com edema na derme e grande quantidade de bastonetes bas?filos. Os demais camundongos foram eutanasiados e n?o tiveram altera??es. A identifica??o por Rea??o em Cadeia de Polimerase (PCR) do meio de Tarozzi do f?gado dos camundongos que morreram foi detectado Clostridium septicum.
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Efeitos da ?gua de coco hipert?nica e sinvastatina no tratamento da sepse e choque hipovol?mico em ratosMedeiros, Vanessa de F?tima Lima de Paiva 28 July 2017 (has links)
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Previous issue date: 2017-07-28 / Reanima??o e tratamento de choque e sepse s?o controversos e desafiadores. O objetivo deste estudo foi examinar os efeitos da ?gua de coco + NaCl 3% + sinvastatina na reanima??o em modelo murino de sepse abdominal e choque hemorr?gico. Ratos Wistar machos adultos foram submetidos a sepse por ligadura e pun??o do ceco e choque hemorr?gico, e divididos em quatro grupos de reposi??o hidroeletrol?tica. 1. solu??o salina 0,9%; 2. ?gua de coco + NaCl 3%; 3. ?gua de coco + NaCl 3% + sinvastatina (microemuls?o) 10 mg / Kg i.v; 4. ?gua de coco n?o modificada. Escore cl?nico, histologia e fluoresc?ncia do pulm?o foram avaliados neste estudo. Dosagem s?rica das citocinas TNF-?, IL-1? e contagem de leuc?citos foram determinados. O tratamento com ?gua de coco + NaCl 3% + sinvastatina resultou em resultados significativamente melhores de sobreviv?ncia, an?lises bioqu?micas e histopatologia quando comparando com os animais submetidos aos tratamentos ?gua de coco + NaCl 3% e ?gua de coco n?o modificada. Em conclus?o, a ressuscita??o com ?gua de coco com Nacl 3% + sinvastatina teve efeito ben?fico significativo sobre a redu??o dos n?veis de citocinas e na atenua??o de les?es pulmonares em modelo de sepse abdominal e choque hemorr?gico. Tamb?m foi demonstrado que o tratamento com ?gua de coco com Nacl 3% + simvastatina atuou positivamente na fun??o do f?gado e dos rins, e no escore cl?nico. / Resuscitation and treatment of shock and sepsis are controversial and challenging. The aim of this study was to examine the effects of coconut water + Nacl 3%+ simvastatin in resuscitation in a murine model of sepsis and hemorrhagic shock. In methods, adult male Wistar rats were subjected to sepsis by cecal ligation and puncture and hemorrhagic shock assigned to four fluid replenishment groups. Animals received 1. saline 0,9%; 2. coconut water (CW) + NaCl 3%; 3. coconut water + NaCl 3% + simvastatin (microemulsion)10 mg/Kg i.v; 4. Coconut water. Clinical score was determined and lung tissue was assessed for inflammation. Serum cytokines TNF-?, IL-1 ? and leucocytes were measured. The results of this study indicated that coconut water+NaCl 3%+ simvastatin decreased significantly serum cytokines, survival, biochemical analysis and lung histopathological testing when compared to saline, water+NaCl 3% and coconut water. In conclusion, resuscitation with coconut water with Nacl 3%+simvastatin had a significant beneficial effect on downregulating cytokines and decreasing lung injury in a rat model of abdominal sepsis and hemorrhagic shock. We also demonstrated that coconut water with Nacl 3%+simvastatin administration clearly made liver and kidney function better and improved clinical score.
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Avalia??o do efeito do composto tipo heparina isolado do caranguejo Chaceon fenneri na hemostasia e na morte celularAraujo, Raquel Helen Brito de 20 July 2012 (has links)
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Previous issue date: 2012-07-20 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Heparin is a pharmaceutical animal widely used in medicine due to its potent
anticoagulant effect. Furthermore, it has the ability to inhibit the proliferation, invasion
and adhesion of cancer cells to vascular endothelium. However, its clinical
applicability can be compromised by side effects such as bleeding. Thus, the search
for natural compounds with low bleeding risk and possible therapeutic applicability
has been targeted by several research groups. From this perspective, this study aims
to evaluate the hemorrhagic and anticoagulant activities and citotoxic effect for
different tumor cell lines (HeLa, B16-F10, HepG2, HS-5,) and fibroblast cells (3T3) of
the Heparin-like from the crab Chaceon fenneri (HEP-like). The HEP-like was purified
after proteolysis, ion-exchange chromatography, fractionation with acetone and
characterized by electrophoresis (agarose gel) and enzymatic degradation. Hep-like
showed eletroforetic behavior similar to mammalian heparin, and high trisulfated /Nacetylated
disaccharides ratio. In addition, HEP-like presented low in vitro
anticoagulant activity using aPTT and a minor hemorrhagic effect when compared to
mammalian heparin. Furthermore, the HEP-like showed significant cytotoxic effect
(p<0.001) on HeLa, HepG2 and B16-F10 tumor cells with IC50 values of 1000
ug/mL, after incubation for 72 hours. To assess the influence of heparin-like on the
cell cycle in HeLa cells, analysis was performed by flow cytometry. The results of this
analysis showed that HEP-like influence on the cell cycle increasing S phase and
decreasing phase G2. Thus, these properties of HEP-like make these compounds
potential therapeutic agents / A heparina ? um agente farmac?utico amplamente utilizado em medicina
devido ao seu potente efeito anticoagulante. Al?m disso, tem a capacidade de inibir
a prolifera??o, invas?o e ades?o de c?lulas cancerosas ao endot?lio vascular. No
entanto, a sua aplicabilidade cl?nica pode ser comprometida por efeitos secund?rios
tais como hemorragia. Assim, a busca de compostos naturais com baixo risco
hemorr?gico e poss?vel aplicabilidade terap?utica tem sido alvo de v?rios grupos de
pesquisa. A partir desta perspectiva, este estudo visa avaliar as atividades
hemorr?gica, anticoagulante e efeito citot?xico para as diferentes linhagens de
c?lulas tumorais (HeLa, B16-F10, HepG2, HS-5,) e c?lulas de fibroblastos (3T3)
proporcionadas pelo composto tipo heparina obtido do caranguejo Chaceon fenneri.
Dessa forma, o composto foi purificado ap?s prote?lise, cromatografia de troca
i?nica e fracionamento com acetona, e caracterizado por eletroforese em gel de
agarose e degrada??o enzim?tica. O composto em estudo mostrou comportamento
eletrofor?tico semelhante ? heparina de mam?fero, e alta raz?o de propor??o de
dissacar?deos trissulfatado / N-acetilado. Al?m disso, o composto apresentou baixa
atividade anticoagulante in vitro usando aPTT e um efeito hemorr?gico menor
quando comparado com heparina de mam?fero. O composto tipo heparina obtido do
caranguejo Chaceon fenneri mostrou efeito citot?xico significativo (p <0,001) em
c?lulas linhagens de c?lulas tumorais HeLa, HepG2 e B16-F10 com valores de IC50
de 1000 ug / mL, ap?s a incuba??o durante 72 horas. Para avaliar a influ?ncia do
composto sobre o ciclo celular em c?lulas HeLa, foi realizada uma an?lise por
citometria de fluxo. Os resultados desta an?lise mostraram que a influ?ncia do
composto sobre o ciclo celular aumenta a fase S e diminui a fase G2. Assim, essas
propriedades do composto tipo heparina obtido do caranguejo Chaceon fenneri
sugerem este composto como um agente terap?utico em potencial
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Planejamento baseado na estrutura da metaloprotease BPMP-I e avalia??o de tiossemicarbazonas ativas contra a pe?onha da serpente Bothrops pauloensis / Structure-based planning Of BPMP-I metalloprotease and evaluation Of thiosemicarbazones active against The snake venom Bothrops PauloensisFerreira, Francis Barbosa 04 August 2016 (has links)
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Previous issue date: 2016-08-04 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / In this work, semi and thiosemicarbazones selected from the LaDMol-QM library, were used
to study their interactions with a metalloproteinase from the snake Bothrops pauloensis
(BpMP-I) by molecular modelling and enzymatic inhibition assays with the toxin. The
crystalographic structure of BaPI (PDB code: 2W12) was used as a mold to build the 3D
model of BpMP-I by homology modeling. The theorical model of BpMP-I showed good
quality parameters and was used in a subsequent molecular modeling study. The
thiossemicarbazones showed better molecular docking results and in vitro enzymatic
inhibitions assays than semicarbazones. Studies by semi-empirical methods indicate a positive
enthalpy of interaction, suggesting that the enzyme inhibition by these compounds must be a
entropy-driven process. The results were used together to select the LDQM-IN-23 compound
and propose rationally designed modifications to improve the interactions with the toxin. The
study of the catalytic site of BpMP-I showed that there is an adjacent pocket with amino
groups of the peptide bonds available for interaction. All results were used together to design
structural changes, aiming the enhancing of the interaction with toxin. Therefore, was
proposed the insertion of the carboxyl group with different spacers, containing 2 (LDQM-IN-
23b) and 3 methylene groups (LDQM-IN-23c). The docking results and semi-empiric
optimization showed that there was a considerable improvement in the interaction for the
modified compounds. The modified compounds were synthesized and tested for biological
and enzymatic inhibition activity. It was observed that the IC50 values have improved: the
original molecule, LDQM-IN-23 has an IC50 of 3,011 ?M and the modified molecules have
IC50 of 79.12 (LDQM-IN-23b) and 1.77 ?M (LDQM-IN-23c). These molecules were tested
for inhibition of hemorrhagic activity induced by Bothropoidin, a P-III class
metalloproteinase, and by the B. pauloensis whole snake venom. The three molecules can
inhibit the hemorrhagic activity induced by isolated toxin and whole venom, and LDQM-IN-
23c showed higher efficiency compared with the other two, and in a rate of 1:10 (w/w
venom/inhibitor) the inhibition of the hemorrhagic activity was 100%. A molecular docking
study of this lead compound with Snake Venom Metalloproteases (SVMPs) from different
snake species and genera showed that this molecule can effectivelly interact with these
SVMPs. / Neste trabalho, foram utilizadas semi e tiossemicarbazonas, selecionadas na quimioteca do
LaDMol-QM (Dequim-UFRRJ), para o estudo das intera??es destas com o s?tio ativo de uma
metaloprotease da pe?onha da serpente Bothrops pauloensis por modelagem molecular e
ensaios de inibi??o da atividade enzim?tica e biol?gica sobre a toxina. A estrutura
cristalogr?fica de uma metaloprotease (BaPI) complexada com um inibidor (um
peptideomim?tico) (c?digo PDB 2W12) foi utilizada como molde para a constru??o do
modelo 3D da metaloprotease da pe?onha de B. pauloensis (BpMP-I). O modelo 3D te?rico
da BpMP-I, in?dito para esta toxina, apresentou bons par?metros de qualidade, sendo
considerado adequado para estudos de planejamento de ligantes baseado na estrutura. As
tiossemicarbazonas obtiveram melhores resultados, quando comparados com os resultados
das semicarbazonas, tanto para os ensaios de docagem molecular quanto para estudos de
inibi??o da atividade enzim?tica in vitro. Estudos por m?todos semiemp?ricos indicam uma
entalpia de intera??o positiva, sugerindo que a inibi??o enzim?tica por estes compostos deve
ser um processo controlado entropicamente. Os resultados foram utilizados para selecionar o
derivado LDQM-IN-23 e propor modifica??es estruturais planejadas racionalmente, visando
melhorar a intera??o deste com a toxina. O estudo do s?tio catal?tico da metaloprotease
mostrou que esta possui uma cavidade adjacente com grupos amino das liga??es pept?dicas
dispon?veis para intera??o. Foi proposta, ent?o, a inser??o de um grupo carboxilato com
diferentes espa?adores, 2 (LDQM-IN-23b) e 3 grupos metileno (LDQM-IN-23c). Os
resultados de docagem e otimiza??o semi-emp?rica mostraram que houve uma melhora
consider?vel na intera??o dos ligantes modificados, os quais foram sintetizados e testados
para as atividades de inibi??o enzim?tica e biol?gica. Na inibi??o enzim?tica, houve melhora
da CI50 com o aumento do espa?ador. O composto LDQM-IN-23 tem CI50 de 3011,00 ?M e
os compostos modificados possuem a CI50 de 79,12 (LDQM-IN-23b) e 1,77 ?M (LDQM-IN-
23c). Estes compostos foram testados para a inibi??o da atividade hemorr?gica in vivo
induzida pela Botropoidina, uma metaloprotease da classe P-III, e pela pe?onha bruta de B.
pauloensis. Os tr?s compostos conseguiram inibir a atividade hemorr?gica induzida pela
toxina isolada e pela pe?onha, sendo que o composto LDQM-IN-23c mostrou maior
efici?ncia, quando comparado com os outros dois, e para a propor??o de 1:10 (m/m
pe?onha/inibidor) a inibi??o da atividade foi de 100%. Foi realizado um estudo de docagem
deste composto l?der com outras metaloproteases de pe?onha de serpentes (SVMPs ? Snake
Venom Metalloproteinases), de esp?cies e g?neros diferentes, mostrando que este ligante
consegue interagir com outras SVMPs e ? um candidato para inibir a atividade hemorr?gica
de SVMPs presentes na pe?onha, n?o s? de B. pauloensis, mas de outras serpentes
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