Detection and localization of oxidized proteins in muscle cells by fluorescence microscopy     

Astruc T  Marinova P  Labas R  Gatellier P  Santé-Lhoutellier V 《Journal of agricultural and food chemistry》2007,55(23):9554-9558

In meat, no detailed studies on the intracellular distribution of oxidized proteins during oxidative stress have been performed, to our knowledge. Therefore, we used fluorescence microscopy to detect and locate protein carbonyls, oxidation products of basic amino acids, generated in bovine M. Rectus abdominis during either exposition to a chemical free radical generating system, or refrigerated storage, or cooking. The technique consisted of an immunohistochemical detection of carbonyls by reaction with the specific probe DNPH (2,4-dinitrophenylhydrazine) followed by the sequential addition of a first antibody against DNPH-carbonylated proteins and a CY3-labeled secondary antibody. The fluorescence of the CY3 probe increased regularly with level of free radical generating system and storage time. Moreover, an important heterogeneity of carbonyl distribution was observed, with a higher oxidation level at the periphery than inside the muscle cells. Cooking induced fluorescence increase only at the periphery of cells. Specific coloration of collagen by Sirius red showed that collagen was not involved in fluorescence. We can deduce that accumulation of oxidized proteins observed in the cell periphery was linked to membrane protein oxidation and not to connective tissue oxidation. Biochemical assays were performed in parallel on membrane and myofibrillar proteins to provide complementary quantitative data on level of oxidized proteins.  相似文献   

First report of Enterocytozoon bieneusi from dairy cattle in Argentina     

Valeria F. Del Coco  María Alejandra Córdoba  Gladys Bilbao  Pinto de Almeida Castro  Juan Angel Basualdo  Mónica Santín 《Veterinary parasitology》2014,199(1-2):112-115

Fecal specimens were obtained from a total of 70 dairy calves less than two months old on 11 municipalities in Buenos Aires, Argentina. After removal of fecal debris by sieving and sucrose flotation, specimens were subjected to PCR to detect the presence of Enterocytozoon bieneusi. PCR revealed a 14.3% of prevalence for E. bieneusi with 10 positive calves from 7 municipalities. Gene sequence analysis conducted in all samples positives by PCR revealed the presence of six genotypes; four previously reported in cattle as well as humans (D, I, J, and BEB4), one never reported in cattle before but previously reported in humans (EbpC), and one novel genotype (BEB10). These results constitute the first molecular characterization of E. bieneusi in Argentina, and suggest a potential risk of zoonotic transmission in this area.  相似文献   

Morphological Analysis of the Tongue and the Digestive Tube of Saffron Finch (Sicalis flaveola brasiliensis,LINNAUES 1766) Apprehended by CETAS/IBAMA‐PB     

R. A. Silva Siqueira  A. C. L. Luna  T. A. Cavalcanti  R. E. G. Rici  M. A. Miglino  R. R. Guerra 《Anatomia, histologia, embryologia》2014,43(1):1-5

The saffron finch (Sicalis flaveola brasiliensis) is a Passeriforme of the Brazilian wildlife. There are scarcely any morphological studies on it, although it is frequently trafficked for its birdsong abilities. Its peculiarities, such as territorialism and developed syrinx that provides outstanding song, draw attention towards its domestication. Thus, this study aimed to morphologically describe the tongue and digestive tube organs of this species to furnish subsidies for nutritional, clinical and conservation studies. Forty‐one birds from the Wild Animal Screening Center (CETAS)/Brazilian Institute of Environment (IBAMA)/city of Cabedelo, state of Paraíba (PB) were used. Samples were collected, identified and sent to standard light microscopy; samples of proventriculus and gizzard were sent to Scanning Electron Microscopy (SEM). The samples showed stratification similar to that of other domestic and wild birds, confirmed in the scanning electron microscopy; however, they differed in the absence of dermal papillae in the tongue, lack of ingluvial glands and lack of muscular mucosa and sub‐mucosa in the large intestine.  相似文献   

Antiplatelet and Anticoagulant Effects of Diterpenes Isolated from the Marine Alga,Dictyota menstrualis     

Laura de Andrade Moura  Ana Carolina Marqui de Almeida  Thaisa Francielle Souza Domingos  Fredy Ortiz-Ramirez  Diana Negr?o Cavalcanti  Valéria Laneuville Teixeira  André Lopes Fuly 《Marine drugs》2014,12(5):2471-2484

Cardiovascular diseases represent a major cause of disability and death worldwide. Therapeutics are available, but they often have unsatisfactory results and may produce side effects. Alternative treatments based on the use of natural products have been extensively investigated, because of their low toxicity and side effects. Marine organisms are prime candidates for such products, as they are sources of numerous and complex substances with ecological and pharmacological effects. In this work, we investigated, through in vitro experiments, the effects of three diterpenes (pachydictyol A, isopachydictyol A and dichotomanol) from the Brazilian marine alga, Dictyota menstrualis, on platelet aggregation and plasma coagulation. Results showed that dichotomanol inhibited ADP- or collagen-induced aggregation of platelet-rich plasma (PRP), but failed to inhibit washed platelets (WP). In contrast, pachydictyol A and isopachydictyol A failed to inhibit the aggregation of PRP, but inhibited WP aggregation induced by collagen or thrombin. These diterpenes also inhibited coagulation analyzed by the prothrombin time and activated partial thromboplastin time and on commercial fibrinogen. Moreover, diterpenes inhibited the catalytic activity of thrombin. Theoretical studies using the Osiris Property Explorer software showed that diterpenes have low theoretical toxicity profiles and a drug-score similar to commercial anticoagulant drugs. In conclusion, these diterpenes are promising candidates for use in anticoagulant therapy, and this study also highlights the biotechnological potential of oceans and the importance of bioprospecting to develop medicines.  相似文献   

Intra-varietal variation for in vitro plant regeneration in the genus Trifolium     

Sant S. Bhojwani  Kathleen Mullins  Daniel Cohen 《Euphytica》1984,33(3):915-921

Summary Seedlings of Trifolium repens showed considerable variation with regard to the morphology and growth of their calli, and their ability for in vitro differentiation of shoots. One of the lines selected for regeneration in primary callus cultures also showed shoot formation from protoplasts. Somatic embryogenesis in callus cultures of T. pratense and T. arvense occurred only in selected seedling lines. This paper highlights the importance of screening a large number of plants within a cultivar of outbreeding species to achieve reproducible plant regeneration from tissue culture.  相似文献   

Genetic diversity in the parental lines and heterosis of the tropical rice hybrids   总被引：2，自引：0，他引：2  

Weijun Xu  Sant S. Virmani  Jose. E. Hernandez  Leocadio S. Sebastian  Edilberto D. Redoña  Zhikang Li 《Euphytica》2002,127(1):139-148

Genetic diversity within and between the maintainer (B) and restorer (R) lines used in hybrid breeding programs of the Philippine was investigated with information from analysis of pedigree record, quantitative traits and SSR assays. Mean coefficients of coancestry were calculated as 0.11 within R lines, 0.27 within B lines, and 0.04 between R and B lines, indicating greater diversity among R lines than among B lines, and the significant divergence between B and R lines. These results are consistent with those obtained from quantitative trait analysis and SSR marker assays. Relative gene diversity for 37 random SSR markers averaged 0.20 within B lines,0.28 within R lines, and 0.52 between two groups of lines. There were no consistent associations among various genetic diversity measures. Random sets of SSR marker and pedigree based diversity measures had no significant correlation with mid-parent heterosis for grain yield and biomass, indicating that prediction of heterosis for complex traits based on these two genetic diversity estimates is difficult. This revised version was published online in August 2006 with corrections to the Cover Date.  相似文献   

Behavioural and physiological responses of three chicken breeds to pre-slaughter shackling and acute heat stress     

Debut M  Berri C  Arnould C  Guemené D  Santé-Lhoutellier V  Sellier N  Baéza E  Jehl N  Jégo Y  Beaumont C  Le Bihan-Duval E 《British poultry science》2005,46(5):527-535

1. The aim of this study was to compare the behavioural and physiological responses to hanging and acute heat stress in three different chicken breeds. Chicks were obtained from a slow-growing French 'Label Rouge' line (SGL), a fast-growing standard line (FGL) and a heavy line (HL). The SGL, FGL and HL birds were slaughtered at their respective market ages of 12, 6 and 6 weeks, in an attempt to achieve similar body weights. Before stunning, birds were either shackled by their legs on the moving line for 2 min (shackling stress: SH) or placed in a room at 35 degrees C and 60% of humidity for 3.5 h and then shackled for 2 min (acute heat stress plus shackling: H + SH) or subjected to minimal stress by shackling for 10 s before stunning (control group: C). 2. Bird physiological responses to the three pre-slaughter treatments were estimated by measuring blood corticosterone, glycaemia, creatine kinase activity, acid-base status and electrolyte concentration as well as lactate content and glycolytic potential in the breast (Pectoralis major) and thigh (Ilio tibialis) muscles. Behavioural responses to shackling stress were evaluated by measuring wing flapping duration, straightening up attempts and vocalisations. 3. Blood corticosterone was higher in SH and H+SH groups than in the C group, regardless of genotype. The struggling activity on the shackle line differed among chicken breeds. It was more intense and occurred more rapidly after hanging in the SGL birds than in both other breeds. Furthermore, SGL struggling activity was not affected by hanging duration while it increased with hanging duration in FGL and HL birds. 4. Wing flapping duration was negatively correlated with blood pH, bicarbonate concentration and positively correlated with breast muscle lactate content, indicating that struggling stimulated antemortem glycolysis activity in breast muscle. Acute heat stress affected blood Ca2+ and Na+ concentration and increased glycaemia and glycolytic potential of thigh muscle. 5. Both acute heat stress and shackling before slaughter were experienced as stressful events by all types of birds.  相似文献   

Prevalence and genotypes of Giardia duodenalis in post-weaned dairy calves     

Trout JM  Santín M  Greiner E  Fayer R 《Veterinary parasitology》2005,130(3-4):177-183

To determine the prevalence of Giardia genotypes in post-weaned dairy calves, fecal specimens were collected from 3 to 11-month-old dairy calves per farm on two farms in Vermont, New York, Pennsylvania, Maryland, Virginia, North Carolina, and Florida. Specimens cleaned of fecal debris and concentrated using CsCl density gradient centrifugation were stained and examined by immunofluorescence microscopy and also subjected to PCR and DNA sequence analysis. Overall, PCR provided more sensitive detection than IFA. Prevalence of Giardia infection, as detected by PCR ranged from 20% on NC-2 to 81% on VT-2, with an overall prevalence of 52% (237 positive samples out of 456 total). DNA sequence analysis of the 16S rRNA gene revealed 87% of the 237 Giardia isolates were Assemblage E, and 13% were Assemblage A although the prevalence of these genotypes varied greatly from farm to farm, with five farms having no Assemblage A Giardia. Therefore, Assemblage E was present in 45% of all animals tested and Assemblage A was present in 7% of the animals. Thus, while many of the calves were infected with a genotype that is not known to be infectious for humans, post-weaned calves on nine of 14 farms did harbor Assemblage A Giardia. Therefore calves should be considered as a potential source of human infectious cysts in the environment, with some farms representing a much higher risk than others.  相似文献   

Prevalence and age-related variation of Cryptosporidium species and genotypes in dairy calves   总被引：8，自引：0，他引：8  

Santín M  Trout JM  Xiao L  Zhou L  Greiner E  Fayer R 《Veterinary parasitology》2004,122(2):103-117

Fifteen dairy farms in seven states on the east coast of the US were each visited on two consecutive years to determinate the prevalence of Cryptosporidium species in pre-weaned (5 days to 2 months) and post-weaned calves (3-11 months), respectively. After each of 971 fecal specimens collected directly from each calf was sieved and subjected to density gradient centrifugation to remove debris and concentrate oocysts, specimens were examined by immunofluorescence microscopy, and polymerase chain reaction (PCR). For all PCR-positive specimens the 18S rRNA gene of Cryptosporidium was sequenced. Cryptosporidium was identified from all farms. Types of housing appeared to have no influence with regard to prevalence of infection. Of 971 calves, 345 were infected with Cryptosporidium (35.5%), but more pre-weaned calves (253 of 503; 50.3%) than post-weaned calves (92 of 468; 19.7%) were found to be infected. A total of 278 PCR-positive specimens characterized by gene sequencing revealed Cryptosporidium parvum, Cryptosporidium andersoni, and two unnamed Cryptosporidium genotypes Bovine B (AY120911) and deer-like genotype (AY120910). The prevalence of these Cryptosporidium species and genotypes appeared to be age related between pre- and post-weaned calves. C. parvum, the only zoonotic species/genotype, constituted 85% of the Cryptosporidium infections in pre-weaned calves but only 1% of the Cryptosporidium infections in post-weaned calves. These findings clearly demonstrate that earlier reports on the presence and prevalence of C. parvum in post-weaned cattle that were based solely on oocyst morphology must be reassessed using molecular methods to validate species and genotype. This finding also indicates that persons handling or otherwise exposed to calves under 2 months of age are at greater risk of zoonotic infection from Cryptosporidium than the risk of infection from exposure to older calves.  相似文献   

Prevalence of Giardia duodenalis genotypes in pre-weaned dairy calves   总被引：1，自引：0，他引：1  

Trout JM  Santín M  Greiner E  Fayer R 《Veterinary parasitology》2004,124(3-4):179-186

To determine the prevalence of Giardia genotypes in pre-weaned dairy calves, fecal samples were collected from a minimum of 18, 1-7-week-old dairy calves per farm on two farms each in the states of Vermont, New York, Pennsylvania, Maryland, Virginia, North Carolina, and Florida. Samples cleaned of fecal debris and concentrated using CsCl density gradient centrifugation were stained and examined by immunofluorescence microscopy and also subjected to PCR and gene sequence analysis. Prevalence by PCR ranged from 9% on a farm in Pennsylvania to 93% on a farm in Vermont, with an average prevalence for 407 calves on 14 farms of 40%. Gene sequence analysis of the TPI, beta-giardin and 16S rRNA genes revealed 85% of the positive samples to be Assemblage E, while 15% were Assemblage A, although the percentages of these genotypes varied greatly from farm to farm. Some farms had no Assemblage A Giardia. Thus, while a majority of the calves were infected with a genotype that is not known to be infectious for humans, calves on 7 of 14 farms did harbor Assemblage A Giardia. Calves should be considered as a potential source of human infectious cysts in the environment.  相似文献