ABSTRACT
Apoptosis and necrosis are two distinct forms of cell death that can occur in response to different agents and stress conditions. In order to verify if the oxidative stress induced by dietary selenium and vitamin E deficiencies can lead muscle cells to apoptosis, one-day-old chicks were reared using diets differing in their vitamin E (0 or 10 IU/kg) and selenium (0 or 0.15 ppm) supplementation. Chick skeletal muscle tissue was obtained from 28-day-old animals and used to verify apoptosis occurrence based on caspase activity detection and DNA fragmentation. Antioxidant deficiency significantly increased caspase-like activity assessed by the hydrolysis of fluorogenic peptide substrates (Abz-peptidyl-EDDnp) at lambdaexc = 320 nm and lambdaem = 420 nm. Proteolytic activation was not accompanied by typical internucleosomal DNA fragmentation detected by field inversion gel electrophoresis. Although the general caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp(O-Me) fluoromethyl ketone (Z-VAD-fmk) (0 to 80 muM) did not block caspase-like activity when preincubated for 30 min with muscle homogenates, the hydrolyzed substrates presented the same cleavage profile in HPLC (at the aspartic acid residue) when incubated with the purified recombinant enzyme caspase-3. These data indicate that oxidative stress causes caspase-like activation in muscle cells and suggest that cell death associated with exudative diathesis (dietary deficiency of selenium and vitamin E) can follow the apoptotic pathway
Subject(s)
Animals , Apoptosis , Caspases , Muscle, Skeletal , Vitamin E Deficiency , Chickens , DNA Fragmentation , Enzyme Activation , Muscle, SkeletalABSTRACT
Apoptosis and necrosis are two distinct forms of cell death that can occur in response to different agents and stress conditions. In order to verify if the oxidative stress induced by dietary selenium and vitamin E deficiencies can lead muscle cells to apoptosis, one-day-old chicks were reared using diets differing in their vitamin E (0 or 10 IU/kg) and selenium (0 or 0.15 ppm) supplementation. Chick skeletal muscle tissue was obtained from 28-day-old animals and used to verify apoptosis occurrence based on caspase activity detection and DNA fragmentation. Antioxidant deficiency significantly increased caspase-like activity assessed by the hydrolysis of fluorogenic peptide substrates (Abz-peptidyl-EDDnp) at lambda exc = 320 nm and lambda em = 420 nm. Proteolytic activation was not accompanied by typical internucleosomal DNA fragmentation detected by field inversion gel electrophoresis. Although the general caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp(O-Me) fluoromethyl ketone (Z-VAD-fmk) (0 to 80 muM) did not block caspase-like activity when preincubated for 30 min with muscle homogenates, the hydrolyzed substrates presented the same cleavage profile in HPLC (at the aspartic acid residue) when incubated with the purified recombinant enzyme caspase-3. These data indicate that oxidative stress causes caspase-like activation in muscle cells and suggest that cell death associated with exudative diathesis (dietary deficiency of selenium and vitamin E) can follow the apoptotic pathway.
Subject(s)
Apoptosis , Caspases/metabolism , Muscle, Skeletal/cytology , Selenium/deficiency , Vitamin E Deficiency/enzymology , Animals , Apoptosis/genetics , Caspase Inhibitors , Chickens , DNA Fragmentation , Enzyme Activation , Enzyme Inhibitors/pharmacology , Muscle, Skeletal/enzymologyABSTRACT
Descrevem-se os achados anatomopatológicos de um caso de osteopatia hipertrófica pulmonar em um cäo adulto sem raça definida e sem histórico clínico. A necropsia, foi observado um nódulo cutâneo na cabeça. Havia outros nódulos nos pulmöes, nos rins e no coraçäo. Macroscopicamente, estes apresentavam coloraçäo esbranquiçada, consistência firme e diâmetro que variavam de pouco milímetros a 10 centímetros, como foi observado nos pulmöes. Todos os quatro membros apresentavam-se grossos e duros, com acentuada proliferaçäo periosteal simétrica. Histologicamente, na superfície externa da cortical havia trabéculas ósseas entremeadas por tecido medular e a proliferaçäo periosteal caracterizada por tecido ósseo imaturo. Foram observadas várias cavidades na cortical. Concluiu-se pelo diagnóstico de osteopatia hipertrófica, secundária à metástase pulmonar de fibrossarcoma cutânea
Subject(s)
Animals , Male , Adult , Bone and Bones , Dogs , Osteopathic MedicineABSTRACT
Young rabbits on high (0.57%) or low (0.24%) calcium were given an aqueous extract of Solanum malacoxylon (S.m.) leaves (20 g dried leaves/200 ml distilled water) intragastrically at 0, 12 and 36 hours. On bothe diets S.m. induced progressive hypophosphatasemia but serum calcium and phosphorus underwent only minor changes. In rabbits necropsied at 0, 12, 36, 60, 84 and 108 hours, S.m. was shown to have a negative effect on the resorbing osteocytes. With retarded osteocytic osteolysis, osteopetrosis resulted. Further regressive changes in the osteocytes resulted in osteonecrosis which was observed within 12 hours after administration of S.m. extract. The osteonecrosis, combined with retarded apposition, later resulted in osteopenia. It was concluded that the recommended dietary calcium for growing rabbits--about 0.6%--is too high. Whereas the histologic appearance of bone in rabbits fed low calcium was normal, bones from rabbits on high calcium showed retarded resorption and the rabbits had a relative hypophosphatasemia.