Characterisation of the grapevine cultivar Picolit by means of morphological descriptors and molecular markers

The phenotypic and genotypic variability of cv. Picolit, an ancient, female-flower cultivar from north-eastern Italy was investigated by means of ampelographic and ampelometric descriptors and by molecular markers, such as microsatellites and AFLPs. Thirty nine samples were collected from old plants (30-100 years old), which showed some differences in morphology and growth. In two samples (P6 and P7) morphological differences were found. These samples showed a different allelic profile at 18 out of the 21 SSRs analysed and were therefore considered not to belong to the cv. Picolit. Of the remaining samples, 35 gave the same allelic pattern at all SSRs and they were therefore considered ‘true-to-type’ Picolit, whereas two of them (P4 and P8) showed several variations, including extra alleles. One of the possible causes of such differences is chimerism. The AFLP analysis, from which samples P6 and P7 were excluded, enabled screening of a larger portion of the genome and confirmed the differences of the P4 and P8 samples from the remaining ones. P4 and P8 were different from the majority of samples at 13 and 37 AFLP loci respectively. A few further polymorphic bands were recorded in the remaining samples, but they were disregarded since they were not always reproducible. This research confirmed the appreciable somatic stability of SSR markers even in long-lived, vegetatively propagated plants, and the occasional occurrence of solid mutations and chimerisms.

Effect of dietary nitrogen level and source on mRNA expression of urea transporters in the rumen epithelium of fattening bulls

This paper aims to study the effect of the dietary treatments on mRNA expression of urea transporter B (UT-B) and some aquaporins (AQP) in rumen epithelium of Italian Simmental young bulls. Eighty animals allocated to 16 pens were fed from about 500 to 650 kg body weight with four experimental diets, which resulted from the combination of two crude protein levels (125 and 110 g/kg dry matter, diets M and L, respectively) and two nitrogen sources (soybean meal (SBM) or SBM partly replaced by an isonitrogenous mixture of corn and urea; diets −U and +U, respectively). At slaughtering samples of blood and rumen epithelium were collected from six bulls for each diet. Blood samples were analysed for haematological parameters and quantitative PCR was carried out on the mRNA extracted from the rumen epithelium samples. The bulls fed diets M had lower plasma concentrations of aspartate aminotransferase than those receiving diets L (78.9 vs. 88.3 U/l, p = 0.04). Plasma urea was higher (p = 0.03) for diets M and lower for diets +U (2.0 vs. 2.5 and 1.73 vs. 2.00 mmol/l, respectively, in M and L diets, p = 0.04). The effect of dietary treatments on rumen UT expression were limited to AQP3, which was down regulated (p = 0.01) in diets +U. Finally, a high positive correlation (R2 = 0.871) between the expressions of AQP7 and AQP10 was found. In conclusion, the AQP3 appears very responsive to dietary treatments and therefore it is a candidate to be further studied in rumen metabolism experiments. The close relationship between mRNA expression of AQP7 and AQP10 indicates a similar function of these two proteins

In Vitro Infection of Trypanosoma cruzi Causes Decrease in Glucose Transporter Protein-1 (GLUT1) Expression in Explants of Human Placental Villi Cultured under Normal and High Glucose Concentrations

Trypanosoma cruzi, the etiologic Chagas' disease agent, induces changes in protein pattern of the human placenta syncytiotrophoblast. The glucose transporter protein-1 (GLUT1) is the primary isoform involved in transplacental glucose transport. We carried out in vitro assays to determine if T. cruzi infection would induce changes in placental GLUT1 protein expression under normal and high concentration of glucose. Using Western blot and immunohistological techniques, GLUT1 expression was determined in normal placental villi cultured under normal or high concentrations of glucose, with or without in vitro T. cruzi infection, for 24 and 48 hours. High glucose media or T. cruzi infection alone reduced GLUT1 expression. A yet more accentuated reduction was observed when infection and high glucose condition took place together. We inform, for the first time, that T. cruzi infection may induce reduction of GLUT1 expression under normal and high glucose concentrations, and this effect is synergic to high glucose concentrations

Heat stress and feeding behaviour of dairy cows in late lactation

Heat stress is one of the most important problems that dairy cows have to face and the use of cooling systems is becoming more and more important. The first reaction that has the animal to cope with the environmental variations is to modify its behaviour. This study was aimed to investigate the effect of heat stress and a cooling system on the feeding behaviour of Italian Holstein Friesian dairy cows in late lactation. Two experiments were performed. In the first experiment, eight dairy cows were firstly kept 7 d under thermoneutral condition, and then under mild heat stress (temperature humidity index, THI, ranging between 72 and 78) for others 7 d. The second experiment consisted of 8 dairy cows used in a two-period cross-over design where the treatment was the use or not of a sprinkler system for cooling cows under mild heat stress. Cows were equipped with a noseband pressure sensor able to detect rumination and eating time, number of rumination and eating chews, number of rumination boluses and rumination intensity. Heat stress reduced rumination time, number of rumination chews and boluses (p <.05), and tended to reduce the number of eating chews (p <.10). Cooled cows increased rumination and eating time (p <.05), rumination intensity (p <.01), and the number of rumination and eating chews (p <.05). In conclusion, feeding behaviour was deeply influenced even by mild heat stress, which was effectively improved by the use of a sprinkler system.HIGHLIGHTS Mild heat stress reduced rumination time, number of rumination chews and boluses of dairy cows in late lactation Cooling cows with sprinklers was effective in alleviating heat stress in terms of feeding behaviour

Caffeine Inhibits EGF-Stimulated Trophoblast Cell Motility through the Inhibition of mTORC2 and Akt.

Impaired trophoblast invasion is associated with pregnancy disorders such as early pregnancy loss and preeclampsia. There is evidence to suggest that the consumption of caffeine during pregnancy may increase the risk of pregnancy loss; however, little is known about the direct effect of caffeine on normal trophoblast biology. Our objectives were to examine the effect of caffeine on trophoblast migration and motility after stimulation with epidermal growth factor (EGF) and to investigate the intracellular signaling pathways involved in this process. Primary first-trimester extravillous trophoblasts (EVT) and the EVT-derived cell line SGHPL-4 were used to study the effect of caffeine on EGF-stimulated cellular motility using time-lapse microscopy. SGHPL-4 cells were further used to study the effect of caffeine and cAMP on EGF-stimulated invasion of fibrin gels. The influence of caffeine and cAMP on EGF-stimulated intracellular signaling pathways leading to the activation of Akt were investigated by Western blot analysis. Caffeine inhibits both EGF-stimulated primary EVT and SGHPL-4 cell motility. EGF stimulation activates phosphatidylinositol 3-kinase, and Akt and caffeine inhibit this activation. Although cAMP inhibits both motility and invasion, it does not inhibit the activation of Akt, indicating that the effects of caffeine seen in this study are independent of cAMP. Further investigation indicated a role for mammalian target of rapamycin complex 2 (mTORC2) as a target for the inhibitory effect of caffeine. In conclusion, we demonstrate that caffeine inhibits EGF-stimulated trophoblast invasion and motility in vitro and so could adversely influence trophoblast biology in vivo

Mulheres na Engenharia: Quebrando Paradigmas

This paper presents the results and experiences of the project "Women in the Engineering: Breaking Paradigms". It was developed in partnership with the Elementery and High School Presidente Juscelino Kubitschek and extended to technical education at IFSC - Campus São José. The project aimed the awakening for the career of engineering in female gender. The performed activities include motivational workshops of programming and robotics, talks and round tables. Este relato apresenta as experiências e resultados  obtidos com o projeto “Mulheres na Engenharia: quebrando paradigmas” (APROEX N°. 03/2014/PROEX – IFSC-SJ), desenvolvido com a Escola Presidente Juscelino Kubitschek e estendido para o âmbito do IFSC - Câmpus São José. O projeto visou o despertar da vocação para a carreira das engenharias no gênero feminino. Foram realizadas  oficinas  de programação e robótica de caráter motivacional,  além de palestras e mesas redondas de debate.DOI: http://dx.doi.org/10.35700/ca.2017.ano4n6.p74-77.185

Co-occurrence of <i>Dinophysis tripos</i> and pectenotoxins in Argentinean shelf waters

The species Dinophysis tripos is a widely distributed marine dinoflagellate associated with diarrheic shellfish poisoning (DSP) events, which has been recently identified as a pectenotoxin (PTX) producer. In two sampling expeditions carried out during austral autumns 2012 and 2013 along the Argentine Sea (≈38–56° S), lipophilic phycotoxins were measured by tandem mass spectrometry coupled to liquid chromatography (LC–MS/MS) in size-fractionated plankton samples together with microscopic analyses of potentially toxic phytoplankton. PTX-2, PTX-11 and PTX-2sa were recurrently detected in the 50–200 μm fractions, in association to D. tripos. PTX-2 was also widely distributed among the 20–50 μm fractions, mostly related to Dinophysis acuminata. Okadaic acid or its analogs were not detected in any sample. This is the first report of D. tripos related to PTX in the Argentine Sea and the first record of PTX-11 and PTX-2sa for this area. The morphological variability of D. tripos, including the presence of intermediate, small and dimorphic cells, is described. Also, the micro- and mesoplanktonic potential grazers of Dinophysis spp. were explored.Facultad de Ciencias Naturales y Muse

Phospholipase C2 Affects MAMP-Triggered Immunity by Modulating ROS Production

The activation of phosphoinositide-specific phospholipase C (PI-PLC) is one of the earliest responses triggered by the recognition of several microbe-associated molecular patterns (MAMPs) in plants. The Arabidopsis (Arabidopsis thaliana) PI-PLC gene family is composed of nine members. Previous studies suggested a role for PLC2 in MAMP-triggered immunity, as it is rapidly phosphorylated in vivo upon treatment with the bacterial MAMP flg22. Here, we analyzed the role of PLC2 in plant immunity using an artificial microRNA to silence PLC2 expression in Arabidopsis. We found that PLC2-silenced plants are more susceptible to the type III secretion system-deficient bacterial strain Pseudomonas syringae pv tomato (Pst) DC3000 hrcC− and to the nonadapted pea (Pisum sativum) powdery mildew Erysiphe pisi. However, PLC2-silenced plants display normal susceptibility to virulent (Pst DC3000) and avirulent (Pst DC3000 AvrRPM1) P. syringae strains, conserving typical hypersensitive response features. In response to flg22, PLC2-silenced plants maintain wild-type mitogen-activated protein kinase activation and PHI1, WRKY33, and FRK1 immune marker gene expression but have reduced reactive oxygen species (ROS)-dependent responses such as callose deposition and stomatal closure. Accordingly, the generation of ROS upon flg22 treatment is compromised in the PLC2-defficient plants, suggesting an effect of PLC2 in a branch of MAMP-triggered immunity and nonhost resistance that involves early ROS-regulated processes. Consistently, PLC2 associates with the NADPH oxidase RBOHD, suggesting its potential regulation by PLC2

Surgical site infection: An observer-blind, randomized trial comparing electrocautery and conventional scalpel

AbstractAimTo evaluate the incidence of surgical site infection (SSI) based on the type of scalpel used for incisions in the skin and in subcutaneous tissues.MethodsObserver-blind, randomized equivalence clinical trial with two arms (electrocautery versus conventional scalpel) which evaluated 133 women undergoing elective abdominal gynecologic oncology surgery. A simple randomization stratified by body mass index (BMI: 30 kg/m2) was carried out. Women were evaluated at 14 and 30 days following the operation. A multivariate analysis was performed in order to check whether the type of scalpel would be a risk factor for SSI.ResultsGroup arms were balanced for all variables, excepted for surgical time, which was significantly higher in the electrocautery group (mean: 161.1 versus 203.5 min, P = 0.029). The rates of SSI were 7.4% and 9.7%, respectively, for the conventional scalpel and electrocautery groups (P = 0.756). The exploratory multivariate model identified body mass index ≥30 kg/m2 (OR = 24.2, 95% CI: 2.8–212.1) and transverse surgical incision (OR = 8.1, 95% CI: 1.5–42.6) as independent risk factors for SSI. The type of scalpel used in surgery, when adjusted for these variables and the surgery time, was not a risk factor for SSI.ConclusionThis study showed that the SSI rates for conventional scalpel and electrocautery were not significantly different. These results were consistent with others reported in the literature and would not allow a surgeon to justify scalpel choice based on SSI.Trial number: NCT01410175 (Clinical Trials – NIH)

Probing formation of cargo/importin-α transport complexes in plant cells using a pathogen effector

Importin-αs are essential adapter proteins that recruit cytoplasmic proteins destined for active nuclear import to the nuclear transport machinery. Cargo proteins interact with the importin-α armadillo repeat domain via nuclear localization sequences (NLSs), short amino acids motifs enriched in Lys and Arg residues. Plant genomes typically encode several importin-α paralogs that can have both specific and partially redundant functions. Although some cargos are preferentially imported by a distinct importin-α it remains unknown how this specificity is generated and to what extent cargos compete for binding to nuclear transport receptors. Here we report that the effector protein HaRxL106 from the oomycete pathogen Hyaloperonospora arabidopsidis co-opts the host cell's nuclear import machinery. We use HaRxL106 as a probe to determine redundant and specific functions of importin-α paralogs from Arabidopsis thaliana. A crystal structure of the importin-α3/MOS6 armadillo repeat domain suggests that five of the six Arabidopsis importin-αs expressed in rosette leaves have an almost identical NLS-binding site. Comparison of the importin-α binding affinities of HaRxL106 and other cargos in vitro and in plant cells suggests that relatively small affinity differences in vitro affect the rate of transport complex formation in vivo. Our results suggest that cargo affinity for importin-α, sequence variation at the importin-α NLS-binding sites and tissue-specific expression levels of importin-αs determine formation of cargo/importin-α transport complexes in plant cells