Proteomic screen finds pSer/pThr-binding domain localizing Plk1 to mitotic substrates     

Elia AE  Cantley LC  Yaffe MB 《Science (New York, N.Y.)》2003,299(5610):1228-1231

We have developed a proteomic approach for identifying phosphopeptide binding domains that modulate kinase-dependent signaling pathways. An immobilized library of partially degenerate phosphopeptides biased toward a particular protein kinase phosphorylation motif is used to isolate phospho-binding domains that bind to proteins phosphorylated by that kinase. Applying this approach to cyclin-dependent kinases (Cdks), we identified the polo-box domain (PBD) of the mitotic kinase polo-like kinase 1 (Plk1) as a specific phosphoserine (pSer) or phosphothreonine (pThr) binding domain and determined its optimal binding motif. This motif is present in known Plk1 substrates such as Cdc25, and an optimal phosphopeptide containing the motif disrupted PBD-substrate binding and localization of the PBD to centrosomes. This finding reveals how Plk1 can localize to specific sites within cells in response to Cdk phosphorylation at those sites and provides a structural mechanism for targeting the Plk1 kinase domain to its substrates.  相似文献   

Molecular correlates of primate nuclear transfer failures     

Simerly C  Dominko T  Navara C  Payne C  Capuano S  Gosman G  Chong KY  Takahashi D  Chace C  Compton D  Hewitson L  Schatten G 《Science (New York, N.Y.)》2003,300(5617):297

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Cell therapy of alpha-sarcoglycan null dystrophic mice through intra-arterial delivery of mesoangioblasts   总被引：1，自引：0，他引：1  

Sampaolesi M  Torrente Y  Innocenzi A  Tonlorenzi R  D'Antona G  Pellegrino MA  Barresi R  Bresolin N  De Angelis MG  Campbell KP  Bottinelli R  Cossu G 《Science (New York, N.Y.)》2003,301(5632):487-492

Preclinical or clinical trials for muscular dystrophies have met with modest success, mainly because of inefficient delivery of viral vectors or donor cells to dystrophic muscles. We report here that intra-arterial delivery of wild-type mesoangioblasts, a class of vessel-associated stem cells, corrects morphologically and functionally the dystrophic phenotype of virtually all downstream muscles in adult immunocompetent alpha-sarcoglycan (alpha-SG) null mice, a model organism for limb-girdle muscular dystrophy. When mesoangioblasts isolated from juvenile dystrophic mice and transduced with a lentiviral vector expressing alpha-SG were injected into the femoral artery of dystrophic mice, they reconstituted skeletal muscle in a manner similar to that seen in wild-type cells. The success of this protocol was mainly due to widespread distribution of donor stem cells through the capillary network, a distinct advantage of this strategy over previous approaches.  相似文献   

In vitro and ex vivo antihydroxyl radical activity of green and roasted coffee   总被引：3，自引：0，他引：3  

Daglia M  Racchi M  Papetti A  Lanni C  Govoni S  Gazzani G 《Journal of agricultural and food chemistry》2004,52(6):1700-1704

The specific antiradical activity against the hydroxyl radical of the water soluble components in green and dark roasted Coffea arabica and Coffea robusta coffee samples, both in vitro by the chemical deoxiribose assay and ex vivo in a biological cellular system (IMR32 cells), were determined. All the tested coffee solutions showed remarkable antiradical activity. In the deoxiribose assay, all the tested solutions showed similar inhibitory activity (IA%) against the sugar degradation (IA values ranged from 45.2 to 46.9%). In the cell cultures, the survival increase (SI%) ranged from 197.0 to 394.0% with C. robusta roasted coffee being significantly more active than the other samples. The coffee solutions underwent dialysis (3500 Da cutoff membrane) to fraction their components. In both systems, the dialysates (MW < 3500 Da) either from green or roasted coffee, showed antiradical activity, while the only retentates (MW > 3500 Da) from the roasted coffee samples were active. The preparative gel-filtration chromatography of roasted coffee C. robusta dialysate gave three fractions active in the biological system, all containing chlorogenic acid derivatives. The most active fraction was found to be that containing the 5-O-caffeoilquinic acid, which shows a linear relation dose-response ranging from 0.02 to 0.10 mM. The results show that both green and roasted coffee possess antiradical activity, that their more active component is 5-O-caffeoyl-quinic acid, and moreover that roasting process induces high MW components (later Maillard reaction products, i.e., melanoidins), also possessing antiradical activity in coffee. These results could explain the neuroprotective effects found for coffee consumption in recent epidemiological studies.  相似文献   

Motivations and Performance Conditions for Ethnic Entrepreneurship     

Enno Masurel  Peter Nijkamp  Murat Tastan  Gabriella Vindigni 《Growth and change》2002,33(2):238-260

Ethnic entrepreneurship has become a popular concept in a modern multi–cultural society. This paper seeks to offer an overview of the potential of ethnic entrepreneurship for solving inter alia the structural unemployment problems of ethnic groups in cities. There is a general lack of empirical evidence on this issue in the current literature. The present paper aims to fill this gap by addressing in particular the critical success conditions for ethnic entrepreneurs. The focus of the research is on variations in success across three ethnic groups in the Amsterdam area. By means of structured personal interviews with many ethnic entrepreneurs, a systematic qualitative data base was created. The paper sets out to identify empirically the driving forces for business success, such as education or the role of informal networks. The explanatory framework deployed for the identification of these qualitative success factors for distinct ethnic groups is based on a particular, recently developed artificial intelligence method, viz. rough set analysis. This multidimensional classification approach appears to be able to identify various important factors for the motivation and performance of ethnic enterprises. Two major findings emerge from this investigation: (i) performance conditions vary across ethnic groups, and (ii) informal networks are crucial for business success.  相似文献   

Benefits and risks of barefoot harness racing in Standardbred trotters     

Marina Sol  Gabriella Lindgren  Erik Bongcam‐Rudloff  Anna Jansson 《Animal Science Journal》2020,91(1)

There is a lack of research on the benefits and risks of shoeing conditions in harness racing. Thus, our objectives were to: (a) investigate whether velocity times (VT; s/km) are affected by racing unshod (N = 76,932 records on 5,247 horses); (b) determine the potential risks of galloping, being penalized, and disqualification when competing unshod (N = 111,755 records on 6,423 horses); and (c) identify additional environmental factors that affect VT and risks. VT was found to be significantly influenced by shoeing condition (e.g., unshod, shod front, shod hind, or fully shod), but also by sex, age, season, track, track condition, start method, start position, distance, and driver‐horse performance level (p < 2e‐16). The risks of galloping and disqualification were significantly influenced by shoeing condition, sex, age, season, track, start method, start position, or driver‐horse performance level (p ≤ .05). Horses racing unshod had 0.7 s/km lower VT than fully shod horses and showed better performance when racing on neutral tracks during the late summer than horses with other shoeing conditions during the same period. However, racing unshod increased the relative risks of galloping and disqualification by 15%–35% in all seasons. Horses shod only on the hind hooves showed better performance than fully shod horses, without higher risks associated with competing unshod.  相似文献   

In vivo and in vitro propagation and transmission of Toggenburg orbivirus     

Planzer J  Kaufmann C  Worwa G  Gavier-Widén D  Hofmann MA  Chaignat V  Thür B 《Research in veterinary science》2011,91(3):e163-e168

The Toggenburg orbivirus (TOV), a recently discovered virus related to bluetongue virus (BTV), has been identified in goats in Switzerland, Italy and Germany. Isolation of TOV in vitro has not yet been achieved and the transmission mechanisms are still unknown. In the experimental infection of pregnant goats described here, TOV could not be detected in secretion/excretion samples or fetal blood. Material from the goat experiment was used as inoculum for propagating the virus in vitro. To enhance the infectivity of TOV several modified protocols, e.g. pretreatment of the virus with trypsin, polyethylene glycol-mediated infection and lipofection were applied. Isolation of TOV, attempts to infect Culicoides nubeculosus by feeding TOV-positive blood and intracerebral inoculation of newborn mice were unsuccessful. The results of these studies suggest that TOV requires specific but different factors than other BTVs for infection and replication outside of its natural caprine host.  相似文献   

Western European epidemiological survey for parvovirus and coronavirus infections in dogs     

Nicola Decaro  Costantina Desario  Monica Billi  Viviana Mari  Gabriella Elia  Alessandra Cavalli  Vito Martella  Canio Buonavoglia 《Veterinary journal (London, England : 1997)》2011,187(2):195-199

An epidemiological survey for canine parvovirus (CPV) and canine coronavirus (CCoV) infections was conducted in Western Europe. A total of 156 faecal samples were collected from dogs with diarrhoea in Spain (n = 47), Italy (n = 39), France (n = 26), Germany (n = 21), the United Kingdom (n = 8), Belgium (n = 10), and the Netherlands (n = 5). Using molecular assays for virus detection and characterisation, CPV and CCoV were found to be widespread in European dog populations, either alone or in mixed infections. In agreement with previous reports, the original type CPV-2 was shown not to circulate in European dogs. The recently identified virus variant CPV-2c was predominant in Italy and Germany and present at high rates in Spain and France but was not detected in the UK or Belgium. Except for the UK, CCoV genotype I was identified in all European countries involved in the survey, albeit at a lower prevalence rates than CCoV genotype II.  相似文献   

Canine coronavirus induces apoptosis in cultured cells   总被引：1，自引：0，他引：1  

Ruggieri A  Di Trani L  Gatto I  Franco M  Vignolo E  Bedini B  Elia G  Buonavoglia C 《Veterinary microbiology》2007,121(1-2):64-72

Canine coronavirus (CCoV) is widespread in dogs in several countries and causes mild enteric illness evolving to severe enteritis in young pups. In in vitro cultures canine coronaviruses generally induce extensive cell death, however nature of the events leading to cell death remains largely unknown. We analysed the induction of cytopathic effect by CCoV in a canine fibrosarcoma cell line (A-72) in order to characterize the apoptotic effect in homologous cell system. Following CCoV infection A-72 cell line, which is permissive to CCoV, showed reduced growth rate, as detected by MTT assay, a standard colorimetric assay for measuring cellular proliferation, and underwent to apoptotic death. Starting from 24h after CCoV infection, cells morphology appeared dramatically changed, with cells rounding and detachment from culture surface. Morphologic and biochemical features of apoptosis, such as blebbing of the plasma membrane, translocation of phosphatidilserine to cell surface and annexin V positive staining, nuclear fragmentation, apoptotic bodies formation and DNA laddering, were detected in CCoV-infected cells. Propidium iodide staining of infected culture indicated the appearance of hypodiploid DNA peak corresponding to apoptotic cell population. Commonly to other animal coronavirus infection caspase-3 is likely to contribute to the execution phase of apoptosis induced by CCoV in A-72 cells since we found activation of enzymatic activity as well as procaspase-3 activating cleavage. Apoptotic death of infected cells is detrimental as it causes cell and tissue destruction as well as inflammatory responses. Therefore in the case of CCoV associated gastroenteritis, apoptosis of epithelial mucosa cells may be responsible for pathology induced by CCoV infection.  相似文献   

Risk factors for canine neosporosis in farm and kennel dogs in southern Italy     

Paradies P  Capelli G  Testini G  Cantacessi C  Trees AJ  Otranto D 《Veterinary parasitology》2007,145(3-4):240-244

Neosporosis by Neospora caninum causes losses to livestock production through abortion in cattle while, in dogs, it induces neuromuscolar disease. This study investigated neosporosis seroprevalence associated risk factors (including the seropositivity to leishmaniosis) in dogs of southern Italy, determined the prevalence of N. caninum oocyst shedding and examined the relationship between seroprevalence of neosporosis in farm dogs and cattle. Using an inhibition ELISA, 20.9% of 306 dogs had percent inhibition values >10 (indicative of exposure) and farm dogs had a significantly (p<0.001) higher seroprevalence than dogs in a rescue kennel. Whilst N. caninum seroprevalence was associated with increasing age in dogs (p< or =0.01) there was no association between seropositivity for N. caninum and for Leishmania infantum. Oocysts of N. caninum were not detected in faecal samples from 230 dogs including 160 farm dogs. The results indicated that neosporosis infection is common in southern Italy both in dogs and in cattle and that dogs at higher risk of exposure are free-ranging ones living in farms. The lack of correlation between canine seroprevalence for N. caninum and L. infantum assumes a particular significance in an endemic area for leishmaniosis.  相似文献