Application of RFLP analysis of recA, gyrA and rpoS gene fragments for rapid differentiation of Erwinia amylovora from Erwinia strains isolated in Korea and Japan     

Malgorzata Waleron  Krzysztof Waleron  Klaus Geider  Ewa Lojkowska 《European journal of plant pathology / European Foundation for Plant Pathology》2008,121(2):161-172

Restriction fragment length polymorphism (RFLP) analysis of the PCR amplified fragments of recA, gyrA and rpoS genes was applied for the characterization of Erwinia amylovora and Erwinia strains, which cause fire blight and Asian pear blight in orchards. Primers, constructed on the basis of the published recA, gyrA and rpoS gene sequences of Erwinia carotovora, allowed us to amplify DNA fragments for RFLP differentiation of E. amylovora and E. pyrifoliae and finally to distinguish strains within these species and relate them to pear pathogens from Japan. Three to seven restriction endonucleases were applied for RFLP analysis of each gene fragment. The electrophoretic patterns generated after PCR–RFLP for each of the tested genes, were characteristic and specific for each species and allowed their differentiation. The data show that PCR–RFLP analysis of the recA, gyrA and rpoS gene fragments may be considered as a useful tool for the identification and differentiation of E. amylovora and E. pyrifoliae. Almost identical restriction patterns of the analyzed gene fragments indicated a high relationship of E. pyrifoliae strains from Korea and pear pathogens from Japan and a divergence to E. amylovora. For quick and effective differentiation of E. amylovora strains from Erwinia strains from Asia without nucleotide sequencing we recommend the amplification of recA and rpoS gene fragments and digestion of each of them with restriction endonuclease Hin6I.  相似文献   

Cadmium and copper reduce hematopoietic potential in common carp (Cyprinus carpio L.) head kidney     

Elzbieta Kondera  Malgorzata Witeska 《Fish physiology and biochemistry》2013,39(4):755-764

The effects of cadmium and copper on activity of common carp head kidney hematopoietic tissue were evaluated. The fish were subjected to short-term (3 h, Cd-s and Cu-s) or long-term (4 weeks, Cd-l and Cu-l) exposures to 100 % 96hLC₅₀ or 10 % 96hLC₅₀, respectively. Head kidneys were isolated weekly from 5 fish of each group for 4 weeks (post-short-term exposure and during long-term exposure). Percentage of early blast cells among the hematopoietic precursors was calculated. Proliferative and apoptotic activity were evaluated using immunocytochemical staining for proliferating cell nuclear antigen (PCNA) and caspase 3, respectively. Hematopoietic activity was calculated as the ratio of proliferating to apoptotic cells. All metal exposures induced an increase in frequency of early blast cells. The frequency of proliferating (PCNA-positive) cells also significantly increased. A considerable and significant increase in the frequency of apoptotic cells was the most pronounced effect of metal exposures. Both short-term and long-term treatments caused similar effects, but in case of Cd exposures, the reaction was more pronounced. All metal exposures reduced hematopoietic potential of fish measured as the ratio of proliferating to apoptotic precursor cell frequency. However, in all cases, hematopoietic activity was higher than 1 showing that the rate of repair of hematopoietic tissue prevailed over destruction.  相似文献   

利用微卫星标记分析核盘菌遗传多样性   总被引：1，自引：0，他引：1  

段晓莉  李强生  江莹芬  范志雄  胡宝成 《中国油料作物学报》2012,34(3):294-299

利用微卫星(SSR)标记,分析来自欧洲和中国的30个核盘菌菌株的遗传多样性和群体结构。结果表明,3对微卫星引物共扩增出21条清晰的谱带,平均每对引物扩增7条谱带,片段长度为158bp～358bp。根据SSR分析结果,30个核盘菌分离物具有较高的遗传相似性。物种水平的Nei遗传多样性指数为0.139 3,Shannon多样性指数为0.248 8。不同菌株群体的Nei遗传距离都较小,为0.009 6～0.049 6。其中俄罗斯群体和奥地利及英国群体之间的遗传距离最大。从30个菌株的UPGMA聚类分析结果看,核盘菌群体结构与地理来源没有明显的直接关系。许多地理来源相同的菌株分散在不同的组里。仅第三组组内的菌株地理来源一致,均来自于中国,且遗传距离比其他菌株的远。群体遗传分析显示,总群体的基因流比较高（2.111 6）,基因分化系数比较低（0.191 5）。  相似文献   

Correlation of quantified contrast-enhanced power Doppler ultrasonography with immunofluorescent analysis of microvessel density in spontaneous canine tumours     

Ohlerth S  Wergin M  Bley CR  Del Chicca F  Laluhová D  Hauser B  Roos M  Kaser-Hotz B 《Veterinary journal (London, England : 1997)》2010,183(1):58-62

Conventionally, tumour vascularity is assessed invasively by immunofluorescent analysis. Quantified contrast-enhanced power Doppler ultrasound has been used to measure tumour angiogenesis non-invasively in humans and experimental animals. The purpose of this study was to correlate quantified contrast-enhanced power Doppler ultrasound with immunofluorescent results in 45 spontaneous canine tumours. With power Doppler, mean vascularity was high in squamous cell carcinomas, moderate in malignant oral melanomas and low in sarcomas. There was high mean vascularity in squamous cell carcinomas and low mean vascularity in sarcomas and malignant oral melanomas. Although Doppler parameters correlated moderately with microvascular density for all tumours (P=0.004, r=0.4), they did not correlate within histology groups. These analyses show that vascularity differs among canine tumour histology groups. However, dependent on the method used, measurement of tumour vascularity can provide different biological information.  相似文献   

Influence of estrus synchronization of prepubertal gilts on embryo quality   总被引：4，自引：0，他引：4  

Ziecik AJ  Biallowicz M  Kaczmarek M  Demianowicz W  Rioperez J  Wasielak M  Bogacki M 《The Journal of reproduction and development》2005,51(3):379-384

Synchronization and superovulation are commonly used to obtain large numbers of embryos for experimental and practical purposes. This study compared the number, quality, and in vitro development of embryos recovered from gilts following single or double estrus synchronization and superovulation. Prepubertal gilts from the single synchronization group were injected with 1500 I.U. PMSG and 1000 I.U. hCG 72 h later. The double synchronized group of gilts was treated with 750 I.U. PMSG and 500 I.U. hCG 72 h later. After 17 days, 1500 I.U. PMSG followed by 1000 I.U. hCG was administered. Five days after insemination embryos were recovered and cultured for 6 days. Both single and double hormonal stimulation schedules resulted in recovery of elevated numbers of embryos (28.4 and 23.4 vs. 11.3; p<0.01 and p相似文献   

Reevaluation of the University of Wisconsin 2-year protocol for treating canine lymphosarcoma     

Kaiser CI  Fidel JL  Roos M  Kaser-Hotz B 《Journal of the American Animal Hospital Association》2007,43(2):85-92

This retrospective study investigated a population of 96 dogs with newly diagnosed malignant lymphosarcoma that were treated with the commonly used University of Wisconsin-Madison (UW-M) chemotherapy protocol. Pretreatment characteristics were analyzed to determine prognostic factors. Dogs with higher World Health Organization (WHO) stages (including stage IV) and dogs with hypercalcemia were at significantly higher risk of relapse (P=0.018 and P=0.016, respectively). Dose reduction, treatment delays, and prior therapy with cortico-steroids were not associated with clinical outcome. First remission duration of 270 days was similar to historically reported data. Overall survival time of 218 days was much shorter than historical data.  相似文献   

Cementless Total Hip Replacement in 20 Juveniles Using BFX™ Arthroplasty     

Andy Y. Law BVetMed  Malgorzata Bielecki Mag.med.vet.  Sarah Girling Bsc BVSc CertSAS Diplomate ECVS 《Veterinary surgery : VS》2014,43(6):715-725

  相似文献   

Structural features of arabinoxylans extracted with water at different temperatures from two rye flours of diverse breadmaking quality     

Cyran M  Courtin CM  Delcour JA 《Journal of agricultural and food chemistry》2003,51(15):4404-4416

The water extractable (WE) arabinoxylans from two rye flours differing in baking quality were studied following sequential extraction with water at 4, 40, and 100 degrees C. Ammonium sulfate fractionation of the resulting WE fractions and subsequent analysis revealed substantial differences in the structure of the isolated materials. Furthermore, it allowed us to identify the factors contributing to arabinoxylan water extractability. Our results provide compelling evidence for the existence of separate polymers in rye WE arabinoxylans with different substitution degrees, ranging from quantitatively dominating, lowly substituted populations (arabinose to xylose ratio, Ara/Xyl approximately 0.5) to comparatively less abundant, highly substituted analogues (Ara/Xyl approximately 1.3). Generally, arabinoxylan water extractability was governed by the relative proportion of lowly and highly branched structures. A gradually increasing proportion of highly substituted populations was observed from cold to hot WE fractions. This was associated with the lower proportion of monosubstituted xylopyranosyl residues in the backbone, the higher proportion of disubstituted xylopyranosyl residues, and the higher level of substitution with feruloyl residues. Notable differences in the ratio of phenolic compounds to arabinose residues were observed between corresponding polymers isolated from rye flours of high and low baking quality, whereas the differences in their molecular weights were much less pronounced.  相似文献   

Characterization of peppercorn, pepper oil, and pepper oleoresin by vibrational spectroscopy methods     

Schulz H  Baranska M  Quilitzsch R  Schütze W  Lösing G 《Journal of agricultural and food chemistry》2005,53(9):3358-3363

NIR-FT-Raman as well as ATR-IR and NIR spectroscopy in combination with efficient chemometric algorithms was applied for rapid determination of piperine in black and white ground pepper and green whole pepper berries as well as pepper oleoresins. Most of the well-resolved Raman signals detected in the spectra of pepper and the related oleoresins can be assigned to piperine, which is known to be the main pungent principle in these products. On the basis of the specific key bands of piperine, also selective Raman mappings were successfully performed to determine the in situ distribution of the alkaloid in the whole green berry and the dried peppercorn as well. It was found that piperine occurs more or less in the whole perisperm of the green fruit. Furthermore, the content and composition of the volatile fraction in various pepper samples were determined by applying the mentioned vibrational spectroscopy techniques. Whereas only NIRS measurements present sufficient reliability to predict the main essential oil substances directly in ground black and white pepper, all spectroscopy methods applied in this study can be used to analyze individual terpenoids in the hydrodistilled oil. It can be assumed that some of the new, efficient vibrational spectroscopy methods have the potential to replace the standard analysis procedures presently applied for the quality control of peppercorns, pepper extracts, and pepper oil.  相似文献   

Leiomodin is an actin filament nucleator in muscle cells     

Chereau D  Boczkowska M  Skwarek-Maruszewska A  Fujiwara I  Hayes DB  Rebowski G  Lappalainen P  Pollard TD  Dominguez R 《Science (New York, N.Y.)》2008,320(5873):239-243

Initiation of actin polymerization in cells requires nucleation factors. Here we describe an actin-binding protein, leiomodin, that acted as a strong filament nucleator in muscle cells. Leiomodin shared two actin-binding sites with the filament pointed end-capping protein tropomodulin: a flexible N-terminal region and a leucine-rich repeat domain. Leiomodin also contained a C-terminal extension of 150 residues. The smallest fragment with strong nucleation activity included the leucine-rich repeat and C-terminal extension. The N-terminal region enhanced the nucleation activity threefold and recruited tropomyosin, which weakly stimulated nucleation and mediated localization of leiomodin to the middle of muscle sarcomeres. Knocking down leiomodin severely compromised sarcomere assembly in cultured muscle cells, which suggests a role for leiomodin in the nucleation of tropomyosin-decorated filaments in muscles.  相似文献