P130Cas-associated protein (p140Cap) as a new tyrosine-phosphorylated protein involved in cell spreading

Integrin-mediated cell adhesion stimulates a cascade of signaling pathways that control cell proliferation, migration, and survival, mostly through tyrosine phosphorylation of signaling molecules. p130Cas, originally identified as a major substrate of v-Src, is a scaffold molecule that interacts with several proteins and mediates multiple cellular events after cell adhesion and mitogen treatment. Here, we describe a novel p130Cas-associated protein named p140Cap (Cas-associated protein) as a new tyrosine phosphorylated molecule involved in integrin- and epidermal growth factor (EGF)-dependent signaling. By affinity chromatography of human ECV304 cell extracts on a MBP-p130Cas column followed by mass spectrometry matrix-assisted laser desorption ionization/time of flight analysis, we identified p140Cap as a protein migrating at 140 kDa. We detected its expression in human, mouse, and rat cells and in different mouse tissues. Endogenous and transfected p140Cap proteins coimmunoprecipitate with p130Cas in ECV304 and in human embryonic kidney 293 cells and associate with p130Cas through their carboxy-terminal region. By immunofluorescence analysis, we demonstrated that in ECV304 cells plated on fibronectin, the endogenous p140Cap colocalizes with p130Cas in the perinuclear region as well as in lamellipodia. In addition p140Cap codistributes with cortical actin and actin stress fibers but not with focal adhesions. We also show that p140Cap is tyrosine phosphorylated within 15 min of cell adhesion to integrin ligands. p140Cap tyrosine phosphorylation is also induced in response to EGF through an EGF receptor dependent-mechanism. Interestingly expression of p140Cap in NIH3T3 and in ECV304 cells delays the onset of cell spreading in the early phases of cell adhesion to fibronectin. Therefore, p140Cap is a novel protein associated with p130Cas and actin cytoskeletal structures. Its tyrosine phosphorylation by integrin-mediated adhesion and EGF stimulation and its involvement in cell spreading on matrix proteins suggest that p140Cap plays a role in controlling actin cytoskeleton organization in response to adhesive and growth factor signaling.     

Apoptotic cascade initiated by angiotensin II in neonatal cardiomyocytes: role of DNA damage

Angiotensin II contributes to ventricular remodeling by promoting both cardiac hypertrophy and apoptosis; however, the mechanism underlying the latter phenomenon is poorly understood. One possibility that has been advanced is that angiotensin II activates NADPH oxidase, generating free radicals that trigger apoptosis. In apparent support of this notion, it was found that angiotensin II-mediated apoptosis in the cardiomyocyte is blocked by the NADPH oxidase inhibitor diphenylene iodonium. However, three lines of evidence suggest that peroxynitrite, rather than superoxide, is responsible for angiotensin II-mediated DNA damage and apoptosis. First, the inducible nitric oxide inhibitor aminoguanidine prevents angiotensin II-induced DNA damage and apoptosis. Second, based on ligation-mediated PCR, the pattern of angiotensin II-induced DNA damage resembles peroxynitritemediated damage rather than damage caused by either superoxide or nitric oxide. Third, angiotensin II activates p53 through the phosphorylation of Ser15 and Ser20, residues that are commonly phosphorylated in response to DNA damage. It is proposed that angiotensin II promotes the oxidation of DNA, which in turn activates p53 to mediate apoptosis.     

根据孢粉资料确定伏尔加地区弗拉阶和法门阶界线

俄罗斯伏尔加格勒伏尔加地区上泥盆统弗拉阶和法门阶的界线以3种剖面类型为代表。第一种类型的界线剖面广泛分布于俄罗斯地台,但两阶之间有沉积间断。第二种类型剖面地层发育较完整,即Volgogradian层直覆于Livendian层之上。前者以Corbulispora viminea-Geminospora vasjamica孢子组合带为特征。第三种类型剖面存在于前里海(Pre-Caspian)凹陷的西侧,是最近根据孢粉资料确定的;地层剖面不完整,在中弗拉阶的Semilukian层有剥蚀面,上覆以不同时代的(弗拉期或法门期)盖层。     

Coordinate induction of glutathione biosynthesis and glutathione-metabolizing enzymes is correlated with salt tolerance in tomato

The acclimation of reduced glutathione (GSH) biosynthesis and GSH-utilizing enzymes to salt stress was studied in two tomato species that differ in stress tolerance. Salt increased GSH content and GSH:GSSG (oxidized glutathione) ratio in oxidative stress-tolerant Lycopersicon pennellii (Lpa) but not in Lycopersicon esculentum (Lem). These changes were associated with salt-induced upregulation of gamma-glutamylcysteine synthetase protein, an effect which was prevented by preincubation with buthionine sulfoximine. Salt treatment induced glutathione peroxidase and glutathione-S-transferase but not glutathione reductase activities in Lpa. These results suggest a mechanism of coordinate upregulation of synthesis and metabolism of GSH in Lpa, that is absent from Lem.     

A new approach to the auchenorrhyncha (Hemiptera, Insecta) cytogenetics: chromosomes of the meadow spittlebug Philaenus spumarius (L.) examined using various chromosome banding techniques

The karyotype of the meadow spittlebug Philaenus spumarius (L.) was studied using conventional chromosome staining, C- and AgNOR- banding, and fluorescent CMA3- and DAPI- techniques. This is the first report on differential staining of the holocentric chromosomes of Auchenorrhyncha. The karyotype of Ph. spumarius includes 2n = 22 + XX/X0. The autosomal pair 1 is large and carries a gap in every homologue. After silver staining, NORs were revealed in both this chromosome pair and a middle-sized pair, most likely 6 or 7. In spermatocyte meiosis, the majority of bivalents formed one chiasma each. The bivalent 1 showed from 1 to 4 chiasmata, the value of 1 or 2 being prevalent. Two further bivalents also showed two chiasmata in some cells. After C-banding, terminal and interstitial dot-type C-heterochromatic blocks were revealed in the chromosomes. In 4 of 11 studied males, the autosomal pair 1 was polymorphic for an extra segment attached to one of the homologues. The segment consisted of both heterochromatic and euchromatic portions. No defined signals were observed in any chromosome treated with DAPI. After CMA3- staining, bright fluorescent signals were obtained in the NOR-bearing chromosomes, suggesting GC-rich DNA bound to the NORs.     

Empirical evidence for key hosts in persistence of a tick-borne disease

An important epidemiological consequence of aggregated host-parasite associations occurs when parasites are vectors of pathogens. Those hosts that attract many vectors will tend to be the focus of transmission. But to what extent, and can we identify characteristics of these key hosts? We investigated these questions with respect to the host-tick relationship of the yellow-necked mouse, Apodemus flavicollis, a critical host in the maintenance of the zoonotic disease, tick-borne encephalitis. Transmission of the virus occurs when ticks feed in a 'co-feeding' aggregation. Thus, the number and frequency of co-feeding groups provides an estimate of the potential rate of virus transmission. We recorded the spatio-temporal variations in co-feeding on a population of rodents in conjunction with recording individual host characteristics. Using Lorenz curves, we revealed conformation of tick-borne encephalitis transmission potential to the 20/80 Rule, where the 20% of hosts most infested with ticks were accountable for 80% of transmission potential. Hosts in the transmission cohort were identified as the sexually mature males of high body mass. Therefore control efforts targeted at this group would substantially reduce transmission potential compared to non-targeted control of the population, which resulted in a linear reduction in transmission potential. Focusing on the 'wrong' functional group would have little impact upon transmission potential until a considerable proportion of the population had been subject to control. However, individuals can change their functional status over time making it difficult to predict the contribution of these individuals to future transmission.     

Liver Lipid Composition and Antioxidant Enzyme Activities of Spontaneously Hypertensive Rats After Ingestion of Dietary Fats (Fish, Olive and High-Oleic Sunflower Oils)

Hypertension is associated with greater than normal lipoperoxidation and an imbalance in antioxidant status, suggesting that oxidative stress is important in the pathogenesis of this disease. Although many studies have examined the effect of antioxidants in the diet on hypertensión and other disorders, less attention has been given to the evaluation of the role of specific dietary lipids in modulating endogenous antioxidant enzyme status. Previously, we have described that liver antioxidant enzyme activities may be modulated by consumption of different oils in normotensive rats. The purpose of the present study was to examine the effects of feeding different lipidic diets (olive oil, OO, high-oleic-acid sunflower oil, HOSO, and fish oil, FO) on liver antioxidant enzyme activities of spontaneously hypertensive rats (SHR). Plasma and liver lipid composition was also studied. Total triacylglycerol concentration increases in plasma and liver of animals fed on the HOSO and OO diets and decreases in those fed on the FO diet, relative to rats fed the control diet. The animals fed on the oil-enriched diet show similar hepatic cholesterol and phospholipid contents, which are higher than the control group. Consumption of the FO diet results in a decrease in the total cholesterol and phospholipid concentration in plasma, compared with the high-oleic-acid diets. In liver, the FO group show higher levels of polyunsaturated fatty acids (PUFA) of the (n-3) series, in relation to the animals fed on the diets enriched in oleic acid. Livers of FO-fed rats, compared with those of OO- and HOSO-fed rats showed: (i) significantly higher activities of catalase, glutathione peroxidase and Cu/Zn superoxide dismutase; (ii) no differences in the NADPH-cytochrome c reductase activity. The HOSO diet had a similar effect on liver antioxidant enzyme activities as the OO diet. In conclusion, it appears that changes in the liver fatty acid composition due mainly to n-3 lipids may enhance the efficiency of the antioxidant defence system and may yield a benefit in the hypertension status. The two monounsaturated fatty acids oils studied (OO and HOSO), with the same high content of oleic acid, but different content of natural antioxidants, had similar effects on the antioxidant enzyme activities studied.