(Table T1) Geochemistry of calcium carbonate veins from ODP Holes 206-1256C and 206-1256D

Drilling a complete deep crustal section has been a primary yet elusive goal since the inception of scientific ocean drilling. In situ ocean crustal sections would contribute enormously to our understanding of the formation and subsequent evolution of the ocean crust, in particular the interplay between magmatic, hydrothermal, and tectonic processes. Ocean Drilling Program (ODP) Leg 206 was the first of a multileg project to drill an in situ crustal section that penetrated the gabbroic rocks of the Cocos plate (6°44.2'N, 91°56.1'W), which formed ~15 m.y. ago on the East Pacific Rise during a period of superfast spreading (>200 mm/yr) (Wilson, Teagle, Acton, et al., 2003, doi:10.2973/odp.proc.ir.206.2003). During Leg 206, the upper 500 m of basement was cored in Holes 1256C and 1256D with moderate to high recovery rates. The igneous rocks recovered are predominantly thin (10 cm to 3 m) basalt flows separated by chilled margins. There are also several massive flows (>3 m thick), although their abundance decreases with depth in Hole 1256D, as well as minor pillow basalts, hyaloclastites, and rare dikes. The lavas have been slightly (<10%) altered by low-temperature hydrothermal fluids, which resulted in pervasive dark gray background alteration and precipitation of saponite, pyrite, silica, celadonite, and calcium carbonate veins. Here we present a geochemical analysis of the CaCO3 recovered from cores. The compositions of ridge flank fluids within superfast spreading crust will be determined from these data, following the approach of Hart et al. (1994, doi:10.1029/93JB02035), Yatabe et al. (2000, doi:10.2973/odp.proc.sr.168.003.2000), and Coggon et al. (2004, doi:10.1016/S0012-821X(03)00697-6).

Argon, uranium, thorium and lead composition and ages of ODP Leg 180 sites

During Ocean Drilling Program (ODP) Leg 180, 11 sites were drilled in the vicinity of the Moresby Seamount to study processes associated with the transition from continental rifting to seafloor spreading in the Woodlark Basin. This paper presents thermochronologic (40Ar/39Ar, 238U/206Pb, and fission track) results from igneous rocks recovered during ODP Leg 180 that help constrain the latest Cretaceous to present-day tectonic development of the Woodlark Basin. Igneous rocks recovered (primarily from Sites 1109, 1114, 1117, and 1118) consist of predominantly diabase and metadiabase, with minor basalt and gabbro. Zircon ion microprobe analyses gave a 238U/206Pb age of 66.4 ± 1.5 Ma, interpreted to date crystallization of the diabase. 40Ar/39Ar plagioclase apparent ages vary considerably according to the degree to which the diabase was altered subsequent to crystallization. The least altered sample (from Site 1109) yielded a plagioclase isochron age of 58.9 ± 5.8 Ma, interpreted to represent cooling following intrusion. The most altered sample (from Site 1117) yielded an isochron age of 31.0 ± 0.9 Ma, interpreted to represent a maximum age for the timing of subsequent hydrothermal alteration. The diabase has not been thermally affected by Miocene-Pliocene rift-related events, supporting our inference that these rocks have remained at shallow and cool levels in the crust (i.e., upper plate) since they were partially reset as a result of middle Oligocene hydrothermal alteration. These results suggest that crustal extension in the vicinity of the Moresby Seamount, immediately west of the active seafloor spreading tip, is being accommodated by normal faulting within latest Cretaceous to early Paleocene oceanic crust. Felsic clasts provide additional evidence for middle Miocene and Pliocene magmatic events in the region. Two rhyolitic clasts (from Sites 1110 and 1111) gave zircon 238U/206Pb ages of 15.7 ± 0.4 Ma and provide evidence for Miocene volcanism in the region. 40Ar/39Ar total fusion ages on single grains of K-feldspar from these clasts yielded younger apparent ages of 12.5 ± 0.2 and 14.4 ± 0.6 Ma due to variable sericitization of K-feldspar phenocrysts. 238U/206Pb zircon, 40Ar/39Ar K-feldspar and biotite total fusion, and apatite fission track analysis of a microgranite clast (from Site 1108) provide evidence for the existence of a rapidly cooled 3.0 to 1.8 Ma granitic protolith. The clast may have been transported longitudinally from the west (e.g., from the D'Entrecasteaux Islands). Alternatively, it may have been derived from a more proximal, but presently unknown, source in the vicinity of the Moresby Seamount.

(Table 1) Strontium concentration and isotope ratios of carbonates from DSDP Holes 69-504B as well as 83-504B and fish debris samples from Indian and Pacific Ocean sediments

Recent revisions of the geological time scale by Kent and Gradstein (in press) suggest that, on the average, Cretaceous magnetic anomalies are approximately 10 m.y. older than in Larson and Hilde's (1975) previous time scale. These revised basement ages change estimates for the duration of alteration in the ocean crust, based on the difference between secondary-mineral isochron ages and magnetic isochron-crustal ages, from 3 to approximately 13 m.y. In addition to the revised time scale, Burke et al.'s (1982) new data on the temporal variation of 87Sr/86Sr in seawater allow a better understanding of the timing of alteration and more realistic determinations of water/rock ratios during seawater-basalt interaction. Carbonates from all DSDP sites which reached Layer 2 of Atlantic crust (Sites 105, 332, 417, and 418) are deposited within 10-15 m.y. of crustal formation from solutions with 87Sr/86Sr ratios identical to unaltered or contemporaneous seawater. Comparisons of the revised seawater curve with the 87Sr/86Sr of basement carbonates is consistent with a duration of approximately 10-15 m.y. for alteration in the ocean crust. Our preliminary Sr and 87Sr/86Sr data for carbonates from Hole 504B, on 5.9-m.y.-old crust south of the Costa Rica Rift, suggest that hydrous solutions from which carbonates precipitated contained substantial amounts of basaltic Sr. For this reason, carbonate 87Sr/86Sr cannot be used to estimate the duration of alteration at this site. A basalt-dominated alteration environment at Hole 504B is consistent with heat-flow evidence which indicates rapid sediment burial of crust at the Costa Rica Rift, sealing it from access by seawater and resulting in unusually low water/rock ratios during alteration.

An Ex Vivo Model for Studying Hepatic Schistosomiasis and the Effect of Released Protein from Dying Eggs

BACKGROUND: We report the use of an ex vivo precision cut liver slice (PCLS) mouse model for studying hepatic schistosomiasis. In this system, liver tissue is unfixed, unfrozen, and alive for maintenance in culture and subsequent molecular analysis.

METHODS AND FINDINGS: Using thick naive mouse liver tissue and sterile culture conditions, the addition of soluble egg antigen (SEA) derived from Schistosoma japonicum eggs, followed 4, 24 and 48 hrs time points. Tissue was collected for transcriptional analysis and supernatants collected to quantitate liver enzymes, cytokines and chemokines. No significant hepatotoxicity was demonstrated by supernatant liver enzymes due to the presence of SEA. A proinflammatory response was observed both at the transcriptional level and at the protein level by cytokine and chemokine bead assay. Key genes observed elevated transcription in response to the addition of SEA included: IL1-α and IL1-β, IL6, all associated with inflammation. The recruitment of antigen presenting cells was reflected in increases in transcription of CD40, CCL4 and CSF1. Indications of tissue remodeling were seen in elevated gene expression of various Matrix MetalloProteinases (MMP3, 9, 10, 13) and delayed increases in TIMP1. Collagen deposition was significantly reduced in the presence of SEA as shown in COL1A1 expression by qPCR after 24 hrs culture. Cytokine and chemokine analysis of the culture supernatants confirmed the elevation of proteins including IL6, CCL3, CCL4 and CXCL5.

CONCLUSIONS: This ex vivo model system for the synchronised delivery of parasite antigen to liver tissue provides an insight into the early phase of hepatic schistosomiasis, corresponding with the release of soluble proteins from dying schistosome eggs.

Efecto del ejercicio de tiro y arrastre sobre el eje renina-angiotensina-aldosterona y cociente testosterona/cortisol en caballos euhidratados y deshidratados

Introducción. El estado hídrico previo al inicio de un ejercicio (hiperhidratación-deshidratación o hipohidratación y euhidratación) modifica la respuesta neurohumoral. En personas, la deshidratación origina una liberación incrementada de las hormonas implicadas en el mantenimiento de la volemia y del estado hidroelectrolítico, tales como la renina (REN), angiotensina (ANG) y aldosterona (ALD). La testosterona (T) es una hormona anabólica, mientras que el cortisol (C) es una hormona de estrés, con funciones catabólicas. El cociente T/C es un reflejo de la intensidad anabólica/catabólica de un ejercicio y en atletas humanos, se ve afectada por el estado hídrico. Objetivos. 1) Describir las variaciones en las concentraciones de REN, ANG, ALD, T, C y ratio T/C en caballos durante un ejercicio de tiro y arrastre, en relación con otros marcadores de equilibrio hidroelectrolítico y en función de la carga arrastrada; 2) Analizar si la respuesta de estas hormonas al ejercicio difiere entre animales con deshidratación hipertónica y euhidratados; Hipótesis: 1) Que las concentraciones de REN, ANG, ALD, T, C serán más elevadas en los caballos deshidratados que en los euhidratados; 2) Que el ejercicio de tiro y arrastre, particularmente en los animales de mayor peso corporal, inducirá una mayor activación del eje REN, ANG, ALD y una liberación superior de T; 3) Que los animales deshidratados experimentarán un incremento más intenso de estas hormonas en respuesta al ejercicio. Material y métodos. Se han llevado a cabo dos estudios simultáneos. En el estudio I, se han analizado los siguientes parámetros: valor hematócrito (HTO), albúmina (ALB), sodio (Na), potasio (K), cloro (Cl), lactato (LA) y concentraciones séricas de REN, ANG y ALD y en el estudio II, se han determinado las concentraciones de ALB, T, C y ratio T/C. En el estudio I, se han incluido 64 caballos machos, enteros y castrados, divididos en dos grupos según su estado hídrico: grupo control o euhidratado (CTR, n=11) y grupo deshidratado (DH; n=53), con deshidratación hipertónica inducida por restricción de agua y comida. Además, según su peso corporal, los animales se han dividido en 3 categorías de peso: I (<350 kg; n=3 para grupo CTR; n=23 para DH), II (351-450 kg; n=3 para CTR; n=18 para DH) y III (>451 kg; n=5 para CTR; n=12 para DH). En el estudio II, se han incluido los mismos animales, con excepción de la categoría de peso III para el grupo CTR, que solo está constituida por 3 animales, en lugar de 5, al descartar a los machos castrados, debido a la medición de T. Todos los animales, realizaron un ejercicio consistente en recorrer una pista de arena de playa de 60 m de longitud, tirando de un carruaje, con 2, 2,25 y 2,5 veces su peso corporal para las tres categorías de peso. La pista de arena se dividió en cuatro áreas de 15 m, en cada una de las cuales, el animal hizo una parada obligatoria, de duración decidida por el acompañante, y que se incluyó como tiempo de carrera. Los animales eran eliminados si se superan los 5 min de duración. En los dos estudios, se tomaron muestras de sangre en los siguientes tiempos: en reposo, antes del ejercicio (R), dentro del primer minuto tras finalizar el ejercicio (E) y a los 5, 10, 15 y 30 minutos de una recuperación pasiva (5REC, 10REC, 15REC y 30REC). Resultados. Estudio I. La diferencia de peso de los caballos tuvo una influencia mínima en los valores basales y en la respuesta al ejercicio. En reposo, los caballos DH tuvieron valores superiores de HTO, ALB, LA, Na, K y ANG en comparación con los CTR. El ejercicio condicionó una elevación de HTO, ALB, LA y Na, tanto en los caballos CTR como en los DH. Se encontró un incremento de ANG en los caballos DH y de ALD en los caballos CTR. La elevación con el ejercicio experimentada por los parámetros analizados fue de magnitud similar en los grupos CTR y DH, con excepción del LA (incrementó más en DH) y ALD (aumentó más en CTR). Estudio II. El ejercicio dio lugar a un aumento de T en las categorías I y III de los grupos CTR y DH, así como un descenso en la categoría II del grupo CTR. El C mostró un aumento progresivo en las tres categorías y en los dos grupos de estado hídrico, con los valores máximos en el tiempo 30REC. El ratio T/C aumentó en la categoría III del grupo DH, descendió en la categoría II del grupo CTR y no varió en los otros casos. Los animales DH, independientemente de la categoría de peso, tuvieron valores superiores de T y C, en la mayoría de los tiempos de extracción de muestras. Sin embargo, si se consideran los cambios experimentados por estas hormonas durante el ejercicio o en la recuperación, en relación a los valores basales, el aumento del C fue más marcado en el grupo CTR que en el DH, para las tres categorías de peso. Las variaciones de la concentración de T fueron superiores en las categorías de peso I y III del grupo CTR que en el grupo DH. El ratio T/C fue estadísticamente igual entre caballos CTR y DH en las categorías I y III, mientras que en la categoría II, los caballos DH presentaron cocientes T/C mayores. Conclusiones. La deshidratación hipertónica altera la concentración de las hormonas implicadas en la regulación del equilibrio hidro-electrolítico, de la presión sanguínea y del metabolismo. Sin embargo, el estado hídrico previo al ejercicio tiene una acción mínima sobre la magnitud de estos cambios en respuesta a un ejercicio de tiro y arrastre de corta duración (inferior a 5 min). Relevancia. Se ha demostrado que la respuesta neurohumoral, determinada en base a las concentraciones circulantes de las hormonas REN, ANG, ALD, T y C, se ve afectada por el ejercicio cuando el animal empieza el mismo en estado de euhidratación o de deshidratación.

Soil enzyme activities and biochemical indexes to assess soil quality in agronomic and forested ecosystems

The soil carries out a wide range of functions and it is important study the effects of land use on soil quality in order to provide most sustainable practices. Three fields trial have been considered to assess soil quality and functionality after human alteration, and to determine the power of soil enzymatic activities, biochemical indexes and mathematical model in the evaluation of soil status. The first field was characterized by conventional and organic management in which were tested also tillage effects. The second was characterized by conventional, organic and agro-ecological management. Finally, the third was a beech forest where was tested the effects of N deposition on soil organic carbon sequestration. Results highlight that both enzyme activities and biochemical indexes could be valid parameters for soil quality evaluation. Conventional management and plowing negatively affected soil quality and functionality with intensive tillage that lead to the downturn of microbial biomass and activity. Both organic and agro-ecological management revealed to be good practices for the maintenance of soil functionality with better microbial activity and metabolic efficiency. This positively affected also soil organic carbon content. At the eutrophic forest, enzyme activities and biochemical indexes positively respond to the treatments but one year of experimentation resulted to be not enough to observe variation in soil organic carbon content. Mathematical models and biochemical indicators resulted to be valid tools for assess soil quality, nonetheless it would be better including the microbial component in the mathematical model and consider more than one index if the aim of the work is to evaluate the overall soil quality and functionality. Concluding, the forest site is the richest one in terms of organic carbon, microbial biomass and activity while, the organic and the agro-ecological management seem to be the more sustainable but without taking in consideration the yield.

Characterization of the chestnut germoplasm of Emilia-Romagna region and ancient fruit tree varieties valorization

Molecular characterization represents a valid support for the recovery of germoplasm, also motivated by the interest for the valorization of local productions in order to make their traceability possible. Molecular characterization is also fundamental for the individuation of misnomers in collection fields in which the different varieties are preserved. In particular, microsatellites have been used in this research to investigate the genetic diversity, inside a population and at an individual level, and the correct varietal correspondence. The research is mainly based on the study of European chestnut (Castanea sativa Mill.) cultivars to evaluate the genetic diversity and relationships in Emilia-Romagna region (Italy). A STRUCTURE analysis was carried out at European level with the allelic frequencies of the samples collected in Emilia-Romagna. Variation found at group and subgroup level may reflect a combination of historical migration/selection processes and adaptive factors to different environments between Italian and Spanish regions. In addition, a case study for the valorization of an old local variety and its re-introduction in the cultivation areas was proposed. This research was carried out by a morphological and molecular characterization of the local apple variety 'Rosa Romana'. The conservation of this variety entails the discrimination of different accessions with very similar phenotype that are present in the original cultivation area. The identification of historical trees and most adequate reference plants are fundamental steps for the correct propagation of this old variety and for the development of nursery activities. This will also promote and re-evaluate the exploitation and protection of such ancient Italian apple cultivars. This model could be in future also carried out for chestnut varieties. In conclusion, analysis with molecular markers is of fundamental importance for the protection and the maintenance of local and ancient varieties which allow to increase the allelic variability available for breeding programs.

A phase II, single arm study of CarbopLatin plus Etoposide with Bevacizumab and Atezolizumab in patients with exTEnded-disease small-cell lung cancer (SCLC) – CeLEBrATE trial

Small cell lung cancer (SCLC) is an aggressive neuroendocrine tumor diagnosed at extended disease SCLC (ES-SCLC) stage in about 70% of cases. The new standard of treatment for patients with ES-SCLC is a combination of platinum-etoposide chemotherapy and atezolizumab or durvalumab, two programmed cell death ligand 1 (PD-L1) inhibitory monoclonal antibodies (mAb). However, the benefit derived from the addition of PD-L1 inhibitors to chemotherapy in ES-SCLC was limited and restricted to a subset of patients. The vascular endothelial growth factor (VEGF) is the most important pro-angiogenic factor implicated in cancer angiogenesis, which is abundant in SCLC and associated with poor prognosis. Antiangiogenic agents, such as bevacizumab, a humanized mAb against VEGF, added to platinum-etoposide chemotherapy improved progression-free survival in SCLC in two trials, but it did not translate into a benefit in overall survival. Nevertheless, VEGF has also acts as a mediator of an immunosuppressive microenvironment and its inhibition can revert the immune-suppressive tumor microenvironment and potentially enhance the efficacy of immunotherapies. Based on available preclinical data, we hypothesized that VEGF inhibition by bevacizumab could improve atezolizumab efficacy in a synergistic way and designed a phase II single-arm trial of bevacizumab in combination with carboplatin, etoposide, and atezolizumab as first-line treatment in ES-SCLC. The trial, which is still ongoing, enrolled 53 patients, including those with treated or untreated asymptomatic brain metastases (provided criteria are met), who received atezolizumab, bevacizumab, carboplatin and etoposide for 4-6 cycles (induction phase), followed by maintenance with atezolizumab and bevacizumab for a maximum of 18 total cycles or until disease progression, patient refusal, unacceptable toxicity. The evaluation of efficacy of the experimental combination in terms of 1-year overall survival rate is not yet mature (primary objective of the trial). The combination was feasible and the toxicity profile manageable (secondary objective of the trial).

Cytotoxic Granules In Distinct Subsets Of Cutaneous Lupus Erythematosus.

In cutaneous lupus erythematosus (CLE), the pathogenetic role of cytotoxic granules has been demonstrated in the subacute and discoid subtypes, which show interface dermatitis, but little is known about tumid (T)CLE, which does not show this interface dermatitis, and evolves with minimal epidermal changes. We studied cytotoxic T lymphocytes and cytotoxic granules in discoid (n = 21), subacute (n = 17), and tumid (n = 21) CLE samples. Skin sections were immunohistochemically stained for CD8, CD56, perforin, granzyme A, granzyme B, and granulysin. Inflammatory cells containing the four subtypes of cytotoxic granules were found in all the three CLE forms; however, only the TCLE group showed a positive correlation between the density of CD8+ cells and each subtype of cytotoxic granule-positive cells. In addition, only the TCLE group showed synergy between the densities of cells containing cytotoxic granule subtypes. Cytotoxic granules are important in the pathomechanism of TCLE. They may perform functions other than apoptosis, including maintenance of inflammation and dermal mucinous deposits in TCLE.

Down-regulation Of Slc8a1 As A Putative Apoptosis-evasion Mechanism By Modulation Of Calcium Levels In Penile Carcinoma.

The SLC8A1 gene, which encodes the Na(+)/Ca(2+) exchanger, plays a key role in calcium homeostasis. Our previous gene expression oligoarray data revealed SLC8A1 underexpression in penile carcinoma (PeCa). The aim of this study was to investigate whether the dysregulation of SLC8A1 expression is associated with apoptosis and cell proliferation in PeCa, via modulation of calcium concentration. The underlying mechanisms of SLC8A1 underexpression were also explored, focusing on copy number alteration and microRNA. Transcript levels of SLC8A1 gene and miR-223 were evaluated by quantitative PCR, comparing PeCa samples with normal glans tissues. SLC8A1 copy number was evaluated by microarray-based comparative genomic hybridization (array-CGH). Caspase-3 and Ki-67 immunostaining, as well as calcium distribution by Laser Ablation Imaging Inductively Coupled Plasma Mass Spectrometry [LA(i)-ICP-MS], were investigated in both normal and tumor samples. Confirming our previous data, SLC8A1 underexpression was detected in PeCa samples (P=0.001) and was not associated with gene copy number loss. In contrast, overexpression of miR-223 (P=0.002) was inversely correlated with SLC8A1 (P=0.015, r=-0.426), its putative repressor. In addition, SLC8A1 underexpression was associated with decreased calcium distribution, high Ki-67 and low caspase-3 immunoexpression in PeCa when compared with normal tissues. Down-regulation of the SLC8A1 gene, most likely mediated by its regulator miR-223, can lead to reduced calcium levels in PeCa and, consequently, to suppression of apoptosis and increased tumor cell proliferation. These data suggest that the miR-223-NCX1-calcium-signaling axis may represent a potential therapeutic approach in PeCa.

Processos adaptativos do tecido muscular esqueletico e tecido conjuntivo : repercussões sobre a flexibilidade

Universidade Estadual de Campinas. Faculdade de Educação Física

Interaction between staining and degradation of a composite resin in contact with colored foods

Composite resins might be susceptible to degradation and staining when in contact with some foods and drinks. This study evaluated color alteration and changes in microhardness of a microhybrid composite after immersion in different colored foods and determined whether there was a correlation between these two variables. Eighty composite disks were randomly divided into 8 experimental groups (n = 10): kept dry; deionized water; orange juice; passion fruit juice; grape juice; ketchup; mustard and soy sauce. The disks were individually immersed in their respective test substance at 37 ºC, for a period of 28 days. Superficial analysis of the disk specimens was performed by taking microhardness measurements (Vickers, 50 g load for 45 seconds) and color alterations were determined with a spectrophotometer (CINTRA 10- using a CIEL*a*b* system, 400-700 nm wavelength, illuminant d65 and standard observer of 2º) at the following times: baseline (before immersion), 1, 7, 14, 21 and 28 days. Results were analyzed by ANOVA and Tukey's test (p < 0.05). Both variables were also submitted to Pearson's correlation test (p < 0.05). The passion fruit group underwent the greatest microhardness change, while the mustard group suffered the greatest color alteration. Significant positive correlation was found between the two variables for the groups deionized water, grape juice, soy sauce and ketchup. Not all color alteration could be associated with surface degradation.

The metal content of bulge field stars from FLAMES-GIRAFFE spectra - I. Stellar parameters and iron abundances

Aims. We determine the iron distribution function (IDF) for bulge field stars, in three different fields along the Galactic minor axis and at latitudes b = -4 degrees, b = -6 degrees, and b = -12 degrees. A fourth field including NGC 6553 is also included in the discussion. Methods. About 800 bulge field K giants were observed with the GIRAFFE spectrograph of FLAMES@VLT at spectral resolution R similar to 20 000. Several of them were observed again with UVES at R similar to 45 000 to insure the accuracy of the measurements. The LTE abundance analysis yielded stellar parameters and iron abundances that allowed us to construct an IDF for the bulge that, for the first time, is based on high-resolution spectroscopy for each individual star. Results. The IDF derived here is centered on solar metallicity, and extends from [Fe/H] similar to -1.5 to [Fe/H] similar to + 0.5. The distribution is asymmetric, with a sharper cutoff on the high-metallicity side, and it is narrower than previously measured. A variation in the mean metallicity along the bulge minor axis is clearly between b = -4 degrees and b = -6 degrees ([Fe/H] decreasing similar to by 0.6 dex per kpc). The field at b = -12 degrees. is consistent with the presence of a gradient, but its quantification is complicated by the higher disk/bulge fraction in this field. Conclusions. Our findings support a scenario in which both infall and outflow were important during the bulge formation, and then suggest the presence of a radial gradient, which poses some challenges to the scenario in which the bulge would result solely from the vertical heating of the bar.

Activation of endogenous p53 by combined p19Arf gene transfer and nutlin-3 drug treatment modalities in the murine cell lines B16 and C6

Background: Reactivation of p53 by either gene transfer or pharmacologic approaches may compensate for loss of p19Arf or excess mdm2 expression, common events in melanoma and glioma. In our previous work, we constructed the pCLPG retroviral vector where transgene expression is controlled by p53 through a p53-responsive promoter. The use of this vector to introduce p19Arf into tumor cells that harbor p53wt should yield viral expression of p19Arf which, in turn, would activate the endogenous p53 and result in enhanced vector expression and tumor suppression. Since nutlin-3 can activate p53 by blocking its interaction with mdm2, we explored the possibility that the combination of p19Arf gene transfer and nutlin-3 drug treatment may provide an additive benefit in stimulating p53 function. Methods: B16 (mouse melanoma) and C6 (rat glioma) cell lines, which harbor p53wt, were transduced with pCLPGp19 and these were additionally treated with nutlin-3 or the DNA damaging agent, doxorubicin. Viral expression was confirmed by Western, Northern and immunofluorescence assays. p53 function was assessed by reporter gene activity provided by a p53-responsive construct. Alterations in proliferation and viability were measured by colony formation, growth curve, cell cycle and MTT assays. In an animal model, B16 cells were treated with the pCLPGp19 virus and/or drugs before subcutaneous injection in C57BL/6 mice, observation of tumor progression and histopathologic analyses. Results: Here we show that the functional activation of endogenous p53wt in B16 was particularly challenging, but accomplished when combined gene transfer and drug treatments were applied, resulting in increased transactivation by p53, marked cell cycle alteration and reduced viability in culture. In an animal model, B16 cells treated with both p19Arf and nutlin-3 yielded increased necrosis and decreased BrdU marking. In comparison, C6 cells were quite susceptible to either treatment, yet p53 was further activated by the combination of p19Arf and nutlin-3. Conclusions: To the best of our knowledge, this is the first study to apply both p19Arf and nutlin-3 for the stimulation of p53 activity. These results support the notion that a p53 responsive vector may prove to be an interesting gene transfer tool, especially when combined with p53- activating agents, for the treatment of tumors that retain wild-type p53.

Characterization of different fruit wines made from cacao, cupuassu, gabiroba, jaboticaba and umbu

The main aim of this work was to produce fruit wines from pulp of gabiroba, cacao, umbu, cupuassu and jaboticaba and characterize them using gas chromatography-mass spectrometry for determination of minor compounds and gas chromatography-flame ionization detection for major compounds. Ninety-nine compounds (C(6) compounds, alcohols, monoterpenic alcohols, monoterpenic oxides, ethyl esters, acetates, volatile phenols, acids, carbonyl compounds, sulfur compounds and sugars) were identified in fruit wines. The typical composition for each fruit wine was evidenced by principal component analysis and Tukey test. The yeast UFLA CA 1162 was efficient in the fermentation of the fruit pulp used in this work. The identification and quantification of the compounds allowed a good characterization of the fruit wines. With our results, we conclude that the use of tropical fruits in the production of fruit wines is a viable alternative that allows the use of harvest surpluses and other underused fruits, resulting in the introduction of new products into the market. (C) 2010 Elsevier Ltd. All rights reserved.