EFFECTS OF RESVERATROL ON VITRIFIED PORCINE OOCYTES

Vitrified MII porcine oocytes are characterized by reduced developmental competence, associated with the activation of the apoptotic pathway. Resveratrol (R), a polyphenolic compound present in several vegetal sources, has been reported to exert, among all its other biological effects, an antiapoptotic one. The aim of this study was to determine the effects of R (2\u2009\ub5M) on the apoptotic status of porcine oocytes vitrified by Cryotop method, evaluating phosphatidylserine (PS) exteriorization and caspases activation. R was added during IVM (A); 2\u2009h postwarming incubation (B); vitrification/warming and 2\u2009h postwarming incubation (C); all previous phases (D). Data on PS exteriorization showed, in each treated group, a significantly higher (P<0.05) percentage of live nonapoptotic oocytes as compared with CTR; moreover, the percentage of live apoptotic oocytes was significantly (P<0.05) lower in all R-treated groups relative to CTR. The results on caspase activation showed a tendency to an increase of viable oocytes with inactive caspases in B, C, and D, while a significant (P<0.05) increase in A compared to CTR was recorded. These data demonstrate that R supplementation in various phases of IVM and vitrification/warming procedure can modulate the apoptotic process, improving the resistance of porcine oocytes to cryopreservation-induced damage

Relationship between postpartum uterine involution and biomarkers of inflammation and oxidative stress in clinically healthy mares (Equus caballus)

To test the hypothesis that delayed/impaired uterine involution could be associated with oxinflammation, westudied the progression of the uterine involution in association with some biomarkers of inflammation andoxidative stress in clinically healthy mares (N\ubc26) during early postpartum. The examination of the repro-ductive tract was performed on Days 7 and 21 after foaling. Uterine involution was assessed considering: a) theincrease of the gravid uterine horn diameter (GUHD) compared with diameter recorded before pregnancy duringthe previous breeding season; b) the level of endometrial edema (EE); c) the degree of accumulation of intra-uterinefluid (IUFA); d) the status of the cervix (CS). Inflammation and oxidative stress were studied by measuringserum amyloid A (SAA), cortisol, DHEA, AOPP, protein carbonyl groups, malondialdheyde (MDA) and thiols inplasma on Days 7 and 21. By Day 21 after parturition, a significant improvement (P<0.01) was observed forGUHD and EE; while IUFA increased in six animals. Plasma SAA and DHEA concentrations were higher when theclinical parameters indicated a lower degree of uterine involution. On Day 7, the cortisol/DHEA ratio was lower inanimals with higher degree of EE. Plasma AOPP and MDA concentrations were significantly lower (P<0.05) inanimals with the lower GUHD. On Day 21, plasma MDA concentrations were significantly lower (P<0.05) inanimals with the lower IUFA. Our data suggest that a mild condition of inflammation and oxidative stress occur inmares with delayed/impaired uterine involution

Scinax cruentommus (Anura: Hylidae) in the upper Rio Negro drainage, Amazonas state, Brazil, with the redescription of its advertisement call

Scinax cruentommus (Anura: Hylidae) na bacia do alto Rio Negro, Amazonas, Brasil, com a redescrição do seu canto de anúncio. Nós apresentamos o primeiro registro de Scinax cruentommus no alto Rio Negro, Amazonas, Brasil. Nós também redescrevemos o seu canto de anúncio e fazemos comparações acústicas com os dados disponíveis para outras espécies do Clado de Scinax ruber da bacia amazônica. Com a obtenção de novos dados para S. cruentommus, os parâmetros acústicos suplementaram a sua diagnose congenérica em relação às outras espécies amazônicas do Clado de S. ruber além das características propostas em sua descrição original.Scinax cruentommus (Anura: Hylidae) in the upper Rio Negro drainage, Amazonas state, Brazil, with the redescription of its advertisement call. We provide the first record for Scinax cruentommus from the upper Rio Negro, Amazonas state, Brazil. We also redescribe its advertisement call and make acoustic comparisons with available data for other species of the Scinax ruber Clade in the Amazon Basin. With the assessment of further data on S. cruentommus, the acoustic traits have supplemented its congeneric diagnosis in comparison with the other Amazonian species of the S. ruber Clade besides the features proposed in its original description

Physico-chemical analysis of Albian (Lower Cretaceous) amber from San Just (Spain) : implications for palaeoenvironmental and palaeoecological studies

Amber from a Lower Cretaceous outcrop at San Just, located in the Eastern Iberian Peninsula (Escucha Formation, Maestrat Basin), was investigated to evaluate its physico-chemical properties. Thermogravimetric (TG) and Differential Thermogravimetric (DTG) analyses, infra-red spectroscopy, elemental and C-isotope analyses were performed. Physico-chemical differences between the internal light nuclei and the peripheral darker portions of San Just amber can be attributed to processes of diagenetic alteration that preferentially took place in the external amber border colonized by microorganisms (fungi or bacteria) when the resin was still liquid or slightly polymerized. δ13Camber values of different pieces of the same sample, from the nucleus to the external part, are remarkably homogeneous, as are δ13Camber values of the darker peripheral portions and lighter inner parts of the same samples. Hence, neither invasive microorganisms, nor diagenetic alteration changed the bulk isotopic composition of the amber. δ13C values of different amber samples range from -21.1‰ to -24‰, as expected for C3 plant-derived material. C-isotope analysis, coupled to palaeobotanical, TG and DTG data and infra-red spectra, suggests that San Just amber was exuded by only one conifer species, belonging to either the Cheirolepidiaceae or Aracauriaceae, coniferous families probably living under stable palaeoenvironmental and palaeoecological conditions

Epigallocatechin-3-Gallate (EGCG) Reduces Rotenone Effect on Stallion Sperm-Zona Pellucida Heterologous Binding

Stallion spermatozoa are highly dependent on oxidative phosphorylation for ATP production to achieve normal sperm function and to fuel the motility. The aim of this study was to evaluate the response of equine sperm under capacitating conditions to the inhibition of mitochondrial complex I by rotenone and to test whether epigallocatechin-3-gallate (EGCG), a natural polyphenol component of green tea, could counteract this effect. After 2-h incubation of stallion spermatozoa in modified Tyrode's medium, rotenone (100 nm, 500 nm and 5 \u3bcm) and EGCG (10, 20 and 60 \u3bcm), alone or in combination, did not induce any significant difference on the percentage of viable cells, live sperm with active mitochondria and spermatozoa with intact acrosome. The inhibition of complex I of mitochondrial respiratory chain of stallion sperm with rotenone exerted a negative effect on heterologous ZP binding ability. EGCG at the concentrations of 10 and 20 \u3bcm (but not of 60 \u3bcm) induced a significant increase in the number of sperm bound to the ZP compared with that for control. Moreover, when stallion sperm were treated with rotenone 100 nm, the presence of EGCG at all the concentrations tested (10, 20 and 60 \u3bcm) significantly increased the number of sperm bound to the ZP up to control levels, suggesting that this green tea polyphenol is able to reduce the toxicity of rotenone

ITS-rDNA phylogeny of Colletotrichum spp. causal agent of apple glomerella leaf spot.

Several diseases have affected apple production, among them there is Glomerella leaf spot (GLS) caused by Colletotrichum spp. The first report of this disease in apple was in plants nearby citrus orchards in São Paulo State, Brazil. The origin of this disease is still not clear, and studies based on the molecular phylogeny could relate the organisms evolutionarily and characterize possible mechanisms of divergent evolution. The amplification of 5.8S-ITS (Internal Transcribed Spacer) of rDNA of 51 pathogenic Colletotrichum spp. isolates from apples, pineapple guava and citrus produced one fragment of approximately 600 bases pairs (bp) for all the isolates analyzed. The amplified fragments were cleaved with restriction enzymes, and fragments from 90 to 500bp were obtained. The sequencing of this region allowed the generation of a phylogenetic tree, regardless of their hosts, and 5 isolated groups were obtained. From the "in silico" comparison, it was possible to verify a variation from 93 to 100% of similarity between the sequences studied and the Genbank data base. The causal agent of GLS is nearly related (clustered) to isolates of pineapple guava and to the citrus isolates used as control

Bacterial Biogeography of the Colon in Dogs With Chronic Inflammatory Enteropathy

The intestinal microbiota is believed to play a role in the pathogenesis of inflammatory bowel disease in humans and chronic inflammatory enteropathy (CIE) in dogs. While most previous studies have described the gut microbiota using sequencing methods, it is fundamental to assess the spatial distribution of the bacteria for a better understanding of their relationship with the host. The microbiota in the colonic mucosa of 22 dogs with CIE and 11 control dogs was investigated using fluorescence in situ hybridization (FISH) with a universal eubacterial probe (EUB338) and specific probes for select bacterial groups. The number of total bacteria labeled with EUB338 probe was lower within the colonic crypts of dogs with CIE compared to controls. Helicobacter spp. and Akkermansia spp. were decreased on the colonic surface and in the crypts of dogs with CIE. Dogs with CIE had increased number of Escherichia coli/Shigella spp. on the colonic surface and within the crypts compared to control dogs. In conclusion, the bacterial microbiota in the colonic mucosa differed between dogs with and without CIE, with depletion of the crypt bacteria in dogs with CIE. The crypt bacterial species that was intimately associated with the host mucosa in control dogs was composed mainly of Helicobacter spp.Peer reviewe

Solid state and solution fine tuning of the linear and nonlinear optical properties of (2-pyrene-1-yl-vinyl)pyridine by protonation&#8211;deprotonation reactions

The unexpected and acido-triggered reversible luminescence and nonlinear optical properties of (2-pyrene-1-yl-vinyl)pyridine, a simple and highly transparent chromophore, are studied both in solution and in the solid state. Remarkably, for the first time the acidomodulation of the NLO response of a poled thin film is reported

Gene therapy of thyroid cancer via retrovirally-driven combined expression of human interleukin-2 and herpes simplex virus thymidine kinase

OBJECTIVE AND DESIGN: Based on our clinical experience with combined gene therapy of glioblastoma, we developed a retroviral vector expressing two therapeutic genes (i.e. thymidine kinase of herpes simplex virus, HSV-TK, and interleukin-2, IL-2) and evaluated its efficiency in vitro and in vivo. METHODS: Expression of therapeutic genes in transduced thyroid carcinoma cells was analyzed by real-time RT-PCR. Ganciclovir sensitivity of infected cells was assessed in vitro in thyroid carcinoma cell lines and in vivo in nude mice bearing xenografted thyroid cancers. The combined effect of IL-2/HSV-TK was compared with the effect of IL-2 alone. RESULTS: Expression of therapeutic genes was higher in differentiated than in anaplastic thyroid carcinoma cells. Ganciclovir treatment led to dose- and time-dependent killing of transduced cells in vitro. A bystander effect was demonstrated by using mixtures of infected and non-infected cells. In vivo studies showed a significant reduction of growth and the presence of an inflammatory infiltrate in transduced thyroid tumors expressing IL-2 alone, as compared with non-infected tumors. By using the retroviral vector expressing IL-2/HSV-TK, treatment with ganciclovir led to complete eradication of anaplastic tumors and a >80% reduction of the size of differentiated thyroid carcinomas. Histological analysis of tumor specimens showed extensive necrosis and inflammatory cell infiltrates. The combination of IL-2/HSV-TK plus ganciclovir was significantly more efficient than IL-2 alone in eradicating tumor masses. The bystander effect was also obtained in vivo. CONCLUSIONS: These findings demonstrate the feasibility and efficiency of a combined immunomodulating and suicide gene therapy approach for thyroid carcinomas