Characterization of SCC\u3cem\u3emec \u3c/em\u3e Instability in Methicillin-Resistant \u3cem\u3eStaphylococcus aureus \u3c/em\u3e Affecting Adjacent Chromosomal Regions, Including the Gene for Staphylococcal Protein A \u3cem\u3e(spa)\u3c/em\u3e

Staphylococcal cassette chromosome mec (SCCmec) represents a sequence of clear clinical and diagnostic importance in staphylococci. At a minimum the chromosomal cassette contains the mecA gene encoding PBP2a but frequently also includes additional antibiotic resistance genes (e.g., ermA and aadC; macrolide and aminoglycoside resistance, respectively). Certain regions within SCCmec elements are hot spots for sequence instability due to cassette-specific recombinases and a variety of internal mobile elements. SCCmec changes may affect not only cassette stability but the integrity of adjacent chromosomal sequences (e.g., the staphylococcal protein A gene; spa). We investigated SCCmec stability in methicillin-resistant Staphylococcus aureus (MRSA) strains carrying one of four SCCmec types cultured in the absence of antimicrobial selection over a 3-month period. SCCmec rearrangements were first detected in cefoxitin-susceptible variants after 2 months of passage, and most commonly showed precise excision of the SCCmec element. Sequence analysis after 3 months revealed both precise SCCmec excision and a variety of SCCmec internal deletions, some including extensive adjacent chromosomal loss, including spa. No empty cassettes (i.e., loss of just mecA from SCCmec) were observed among the variants. SCCmec stability was influenced both by internal mobile elements (IS431) as well as the host cell environment. Genotypically similar clinical isolates with deletions in the spa gene were also included for purposes of comparison. The results indicate a role for host-cell influence and the IS431 element on SCCmec stability

Corticosterone Regulates Both Naturally Occurring and Cocaine‐Induced Dopamine Signaling by Selectively Decreasing Dopamine Uptake

Stressful and aversive events promote maladaptive reward‐seeking behaviors such as drug addiction by acting, in part, on the mesolimbic dopamine system. Using animal models, data from our laboratory and others show that stress and cocaine can interact to produce a synergistic effect on reward circuitry. This effect is also observed when the stress hormone corticosterone is administered directly into the nucleus accumbens (NAc), indicating that glucocorticoids act locally in dopamine terminal regions to enhance cocaine\u27s effects on dopamine signaling. However, prior studies in behaving animals have not provided mechanistic insight. Using fast‐scan cyclic voltammetry, we examined the effect of systemic corticosterone on spontaneous dopamine release events (transients) in the NAc core and shell in behaving rats. A physiologically relevant systemic injection of corticosterone (2 mg/kg i.p.) induced an increase in dopamine transient amplitude and duration (both voltammetric measures sensitive to decreases in dopamine clearance), but had no effect on the frequency of transient release events. This effect was compounded by cocaine (2.5 mg/kg i.p.). However, a second experiment indicated that the same injection of corticosterone had no detectable effect on the dopaminergic encoding of a palatable natural reward (saccharin). Taken together, these results suggest that corticosterone interferes with naturally occurring dopamine uptake locally, and this effect is a critical determinant of dopamine concentration specifically in situations in which the dopamine transporter is pharmacologically blocked by cocaine

Investigation on genetically modified soybean (RoundUp Ready) in goat nutrition: DNA detection in suckling kids

The presence of plant DNA fragments in blood, kidney, hearth, liver, spleen and muscle tissue from suckling kids was investigated by using PCR approach. Fragments of high copy number chloroplast and low copy soybean lectin genes were found in several samples of kids whose mother were fed diet containing conventional (control) or transgenic soybean (treated). Only in treated group, fragments of 35S and CP4 epsps soybean genes were found in several samples

In vitro and in vivo inhibition of breast cancer cell growth by targeting the Hedgehog/GLI pathway with SMO (GDC-0449) or GLI (GANT-61) inhibitors.

Aberrant Hedgehog (Hh)/glioma-associated oncogene (GLI) signaling has been implicated in cancer progression. Here, we analyzed GLI1, Sonic Hedgehog (Shh) and NF-κB expression in 51 breast cancer (ductal carcinoma) tissues using immunohistochemistry. We found a positive correlation between nuclear GLI1 expression and tumor grade in ductal carcinoma cases. Cytoplasmic Shh staining significantly correlated with a lower tumor grade. Next, the in vitro effects of two Hh signaling pathway inhibitors on breast cancer cell lines were evaluated using the Smoothened (SMO) antagonist GDC-0449 and the direct GLI1 inhibitor GANT-61. GDC-0449 and GANT-61 exhibited the following effects: a) inhibited breast cancer cell survival; b) induced apoptosis; c) inhibited Hh pathway activity by decreasing the mRNA expression levels of GLI1 and Ptch and inhibiting the nuclear translocation of GLI1; d) increased/decreased EGFR and ErbB2 protein expression, reduced p21- Ras and ERK1/ERK2 MAPK activities and inhibited AKT activation; and e) decreased the nuclear translocation of NF-κB. However, GANT-61 exerted these effects more effectively than GDC-0449. The in vivo antitumor activities of GDC-0449 and GANT- 61 were analyzed in BALB/c mice that were subcutaneously inoculated with mouse breast cancer (TUBO) cells. GDC-0449 and GANT-61 suppressed tumor growth of TUBO cells in BALB/c mice to different extents. These findings suggest that targeting the Hh pathway using antagonists that act downstream of SMO is a more efficient strategy than using antagonists that act upstream of SMO for interrupting Hh signaling in breast cancer

Breast meat traits of Muscovy ducks fed on a microalga (Crypthecodinium cohnii) meal supplemented diet

A trial was conducted in order to increase the docosahexaenoic acid ( DHA) content in the meat of Muscovy ducks ( Cairina moschata domestica L.) fed on a diet supplemented with the microalga Crypthecodinium cohnii. 2. Two diets were provided to 48 male and 48 female ducks, belonging to an Italian rural strain during the last 3 weeks of life: a maize - soybean based diet as the control diet and the same diet supplemented with 5 g/ kg microalga meal. 3. Dietary treatment did not induce differences in growth performances and slaughter traits. Similarly, chemical composition, colour, pH, oxidative stability and sensory characteristics of breast muscle were not influenced by the diet. 4. A significant increase of DHA content in breast meat of ducks fed on the Crypthecodinium cohnii enriched diet was observed

Cryopreservation of pheasant semen: effect of dilution ratio and cooling time on spermatozoa viability and mobility

Aim of the present study was to investigate the cryopreservation of pheasant semen by adopting the freezing/thawing protocol by Tselutin et al. (1999) with some modifications. Semen was collected by abdominal massage twice a week. Evaluations were performed on pooled semen from fifteen males (Phasianus colchicus mongolicus). Two semen dilutions (DIL) with Lake diluent (1:2 and 1:3, v/v; Lake, 1968) and two equilibration times (ET) at 5°C (10min and 30min) before dimethylacetamide (DMA) addition, were tested. Assessment of sperm mobility was performed by Accudenz methodology according to Froman’s procedure (1997) and viability by eosin/nigrosin staining. As expected, viability and mobility were strongly affected by the freezing/thawing procedure. ET did not affect mobility while influenced live sperm percentage during the DMA equilibration (DMA-Eq). Semen/diluent ratio significantly (p<0.001) modified the mobility parameter and the highest progressive movements of spermatozoa were obtained with the most diluted semen in each critical step of the cryopreservation procedure. In conclusion, for pheasant semen cryopreservation, the 1:3 dilution ratio can be considered appropriate and the cooling time up to 30 minutes not crucial for the spermatozoa mobility and viability. Nevertheless, the deleterious effect of freezing/thawing procedure reduced live thawed spermatozoa to 24% and forward motility to 89% of the initial movement capacity

A Very Hot, High Redshift Cluster of Galaxies: More Trouble for Omega_0 = 1

We have observed the most distant (z=0.829) cluster of galaxies in the Einstein Extended Medium Sensitivity Survey, with the ASCA and ROSAT satellites. We find an X-ray temperature of 12.3 +3.1/-2.2 keV for this cluster, and the ROSAT map reveals significant substructure. The high temperature of MS1054-0321 is consistent with both its approximate velocity dispersion, based on the redshifts of 12 cluster members we have obtained at the Keck and the Canada-France-Hawaii telescopes, and with its weak lensing signature. The X-ray temperature of this cluster implies a virial mass ~ 7.4 x 10^14 h^-1 solar masses, if the mean matter density in the universe equals the critical value, or larger if Omega_0 < 1. Finding such a hot, massive cluster in the EMSS is extremely improbable if clusters grew from Gaussian perturbations in an Omega_0 = 1 universe. Combining the assumptions that Omega_0 = 1 and that the intial perturbations were Gaussian with the observed X-ray temperature function at low redshift, we show that the probability of this cluster occurring in the volume sampled by the EMSS is less than a few times 10^{-5}. Nor is MS1054-0321 the only hot cluster at high redshift; the only two other $z > 0.5$ EMSS clusters already observed with ASCA also have temperatures exceeding 8 keV. Assuming again that the initial perturbations were Gaussian and Omega_0 = 1, we find that each one is improbable at the < 10^{-2} level. These observations, along with the fact that these luminosities and temperatures of the high-$z$ clusters all agree with the low-z L_X-T_X relation, argue strongly that Omega_0 < 1. Otherwise, the initial perturbations must be non-Gaussian, if these clusters' temperatures do indeed reflect their gravitational potentials.Comment: 20 pages, 4 figures, To appear in 1 Aug 1998 ApJ (heavily revised version of original preprint

Quality Of Life And Epilepsy Surgery In Childhood And Adolescence.

Epilepsy can affect the quality of life (QOL) of patients. The temporal lobe epilepsy (TLE) is often refractory to medication, which has an adverse impact on QOL. The surgery can be a form to control the seizures and to improve the QOL of the patients. The aim of this study was to verify the QOL of children and adolescents with TLE who underwent surgery for epilepsy, comparing QOL before and after surgery and investigating which parameters showed improvement. We used semi-structured questionnaire in the pre-and post-surgery in 13 patients. The data were analyzed using the Wilcoxon test. The analysis showed that there was general improvement in the QOL postoperatively. There was improvement in general health issues, adverse effects of antiepileptic drugs and the relationship with parents. When properly indicated, epilepsy surgery improves quality of life of patients with TLE.6923-