Experimental left pneumonectomy in pigs. Procedure and management

Background Because there is no detailed description of procedures and perioperative management of major pulmonary resections in swine, we reviewed our experience to delineate the most effective practice in performing left pneumonectomy. Materials and methods Analysis of 11 consecutive left pneumonectomies. Animal data, operative reports, anesthesia records, and perioperative facts were evaluated. Follow-up information until postoperative day 60, methods of care-taking, therapy administration, and all the stabling aspects were systematically assessed. The investigation was aimed at highlighting those procedural steps or details which make the difference in optimizing the available resources (animals, instruments, and personnel). No statistical analysis was performed considering data characteristics and the descriptive nature of information. Results Surgery requires a median time of 2 h and 16 min; two operators and one anesthesiologist represent the basic team. Circulators' number depends on goals to accomplish. The most straightforward procedure requires careful dissection of the pulmonary ligament (limited view), pulmonary veins (low variability), pulmonary artery (delicate), and finally bronchus (no variability observed). The key factors for good anesthesia management have been identified: sedation by caregivers, preoxygenation before induction of general anesthesia, high respiratory rates with low tidal volume after pneumonectomy, and noninvasive ventilation after extubation. Antibiotic prophylaxis has been performed. Postoperative care must be continuous until animals are able to stand up, afterward "preventive noncurative," and always animal friendly. Ideas for minimally stressful therapy administration are helpful. Conclusions After the delineation of this methodology, the compliance to a routine practice allowed us to reduce time, stress, and cost; quality and quantity of possible research increased

T-wave axis deviation, metabolic syndrome and cardiovascular risk: results from the MOLI-SANI study

Early recognition of patients at increased cardiovascular risk is a major challenge. The surface electrocardiogram provides a useful platform and it has been used to propose several indexes. T wave axis abnormality is associated with an increased risk of cardiovascular mortality, independently of other risk factors and can be associated with the presence of the metabolic syndrome (MetS). We assessed the prevalence of T axis abnormalities and its relationship with MetS and its components in a large population of Italian adults. Data concerning 11,143 women (54±11years) and 9742 men (55±11years) randomly recruited from a general population (Moli-sani cohort) were analyzed. After excluding subjects with incomplete data and with history of cardiac disease or left ventricular hypertrophy, T-wave axis was normal in 74.5% of men and 80.9% of women, borderline in 23.6% and 17.3% and abnormal in 1.9% and 1.8%. In subjects with MetS, the prevalence of borderline or abnormal T-wave axis deviation was higher than in subjects without MetS (in men: 26.6% vs. 22.1% and 2.5% vs. 1.7%; in women: 25% vs. 15% and 2.4% vs. 1.6%, respectively for borderline and abnormal levels, pb0.0001). Each component of MetS increased the odds of having borderline or abnormal T-wave axis deviation by 1.21 in men and 1.31 in women. T wave axis deviation is associated with MetS and its individual components. These findings confirm previous reported results, expanding them to a large and representative sample of European population of Caucasian ethnicity

T-wave axis deviation, metabolic syndrome and estimated cardiovascular risk in men and women of the MOLI-SANI Study

Aim: We aimed at investigating the association between T-wave axis deviation, metabolic syndrome (MetS), its components and estimated risk of cardiovascular disease (CVD) at 10 years in a adult Italian population. Methods: 11,143 women (54±11 years) and 9,742 men (55±11 years) were analysed from the Molisani cohort, randomly recruited from the general population. MetS was defined using the ATPIII criteria. T-wave axis deviation was measured from the standard 12-lead resting electrocardiogram. CVD risk in ten years was estimated by the CUORE score. Results: 29% of men and 27% of women with MetS showed borderline or abnormal T-wave as compared to 24% and 17% without MetS (p<0.0001 for both genders). Among components of MetS, elevated waist and blood pressure were strongly associated with Twave axis deviation, whereas glucose, HDL and triglycerides were only marginally. The odds of having borderline or abnormal T-wave axis deviation in multivariable regression analysis, was 1.38 (95% CI:1.25-1.53) in MetS men and 1.68 (95% CI:1.51-1.87) in MetS women compared to those without. Further adjustment for MetS components completely abolished the associations. Abnormal T-wave axis deviation was associated with an increased risk of CVD in 10 years in men (OR=4.4; 95% CI:1.10-17.9). Conclusion: T-wave axis deviation is strongly associated with components of the MetS, in particular high waist circumference and blood pressure and with an increased CVD risk, particularly in men. ECG monitoring to identify T-wave axis deviation in obese, hypertensive or MetS subjects can be an early indicator of vascular disease and help in reducing cardiac events

Effect of methyljasmonate and light on stilbene production in cell lines of Vitis vinifera and other Vitis species

Plants are constantly exposed to both biotic and abiotic environmental stress. Plant responses to these stresses involve different defense mechanisms, including the neo-synthesis of secondary metabolites, called fitoalexins (Chang et al. 2016). Among these, particular interest is addressed to the stilbenes, both for the biological role of chemical defense and for the numerous and well-proven biological activities on human health (Sirerol et al. 2015). Our study is focused on the biosynthesis of stilbenes, firstly resveratrol (the main exponent) and viniferins (resveratrol oligomers), in response to biotic stress in several species of the genus Vitis. In the plant, the accumulation of stilbenes is induced by the presence of phytopathogenic fungi (Langcake & Pryce 1976) or by abiotic stimuli as UV radiation (Roldan et al. 2003). Different stilbenes produced by grape show a synergistic activity, furthermore the quali-quantitative composition of the stilbenic content determines the resistance against specific pathogens. The species Vitis vinifera includes several cultivars quite different from the genetic point of view, since they are exclusively agamic propagated. Consequently, each of these show different levels of suscettibility to pathogens, such as Botrytis cinerea and Aspergillus spp., in relation to the different stilbene profile. Recently, great interest has also been paid for species of Vitis popularly known as “american grapes”, introduced in Europe as rootstocks due to their resistance against some important pathogens of V. vinifera. At present, resistance mechanisms of the american grapes are still unknown. The few avaiable data has been obtained from samples cultivated in field, where the conditions are not fully under control. The aim of the present research is to compare the stilbenic content of the cellular extracts in response to elicitation with methyljasmonate (MeJa), to identify and quantify the main stilbenes involved in resistance against phytopathogenic fungi. The role of light on stilbene production in the same cell cultures will also be investigated. Unespectedly, the obtained results indicate that cultivars of V. vinifera have a richer stilbenic content than the american grapes. For instance, V. vinifera cv. malvasia, in response to MeJa, increases the production of total stilbenes from 2.58 to 15.58 mg/g DW while Vitis rupestris cv. du Lot increases from 2.80 to 8.97 mg/g DW. Moreover, cell lines grown under light conditions accumulate higher levels of total stilbenes, both constitutively and in response to elicitation (e.g. malvasia: from 8.69 to 37.81 mg/g DW). Bibliography • Chang X. et al. (2012) PLoS One 7.7: e40446. • Langcake P. et al. (1976) Physiol Plant Path 9.1: 77-86. • Sirerol J. et al. (2015) Oxid Med Cell Longev 2016:3128951. • Roldán A. et al. (2003) J Agric Food Chem 51.5: 1464-1468

Stilbene biosynthesis and gene expression in response to methyl jasmonate and continuous light treatment in Vitis vinifera cv. Malvasia del Lazio and Vitis rupestris Du Lot cell cultures

Vitis rupestris is used as rootstock or to obtain hybrids with Vitis vinifera, due to its resistance to certain pathogens. Its resistance mechanisms are poorly understood, while it is known that stilbene neo‐synthesis is a central defense strategy in V. vinifera. In the present study, the response to methyl jasmonate (MeJa) and light treatment in terms of stilbene biosynthesis and the expression of genes involved in polyphenol biosynthesis was investigated in V. vinifera and V. rupestris cells. The two species exhibited a similar constitutive stilbene content [2.50–2.80 mg g−1 dry weight (DW)], which greatly increased in response to elicitation (8.97–11.90 mg g−1 DW). In V. vinifera, continuous light treatment amplified the effect of MeJa, with a stilbene production that had never previously been obtained (26.49 mg g−1 DW). By contrast, it suppressed the effect of MeJa in V. rupestris. Gene expression was consistent with stilbene production in V. vinifera, whereas discrepancies were recorded in V. rupestris that could be explained by the synthesis of stilbenes that had never before been analyzed in this species

Poly(lactic-co-glycolic) acid nanoparticles uptake by Vitis vinifera and grapevine-pathogenic fungi

Poly(lactic-co-glycolic) acid (PLGA)- based NPs are currently considered among the most promising drug carriers, nevertheless their use in plants has never been investigated. In this work, for the first time, we demonstrated the ability of PLGA NPs to cross the plant cell wall and membrane of Vitis vinifera cell cultures and grapevine-pathogenic fungi. By means of fluorescence microscopy, we established that PLGA NPs can enter in grapevine leaf tissues through stomata openings and that they can be absorbed by the roots and transported to the shoot through vascular tissues. TEM analysis on cultured cells showed that NPs B 50 nm could enter cells, while bigger ones remained attached to the cell wall. Viability tests demonstrated that PLGA NPs were not cytotoxic for V. vinifera-cultured cells. The cellular uptake of PLGA NPs by some important grapevinepathogenic fungi has also been observed, thus suggesting that PLGA NPs could be used to deliver antifungal compounds within fungal cells. Overall the results reported suggest that such NPs may play a key role in future developments of agrobiotechnologies, as it is currently happening in biomedicine

Stilbene production in response to methyl jasmonate and light in cell cultures of several Vitis species

Stilbenes are secondary metabolites produced by grapevine in response to stress, which play the role of phytoalexins in the plant (1). Recently, stilbenes have received a great deal of attention both for their biological role in the plant and for their well-proven biological activities on human health (1). Plant cell cultures have been investigated for their potential as valuable system for stilbene production (2). Among different elicitors, methyl jasmonate (MeJa) was shown to be the most effective in stimulating stilbene biosynthesis in cell suspension cultures of cultivar Italia (2). In the plant, the accumulation of stilbenes is induced by the presence of phytopathogenic fungi (3) or by abiotic stimuli as UV radiation (4). Different stilbenes produced by grapevine show a synergistic activity, furthermore the quali-quantitative composition of stilbene content determines the resistance against specific pathogens. The species Vitis vinifera includes several cultivars quite different from the genetic point of view, since they are exclusively agamic propagated. Consequently, each of these shows different levels of susceptibility to pathogens, such as Botrytis cinerea, Aspergillus spp. and Plasmopara viticola in relation to the different stilbene profile. Recently, great interest has been also addressed for species of Vitis popularly known as “american grapes”, introduced in Europe as rootstocks due to their resistance against some important pathogens of V. vinifera . At present, resistance mechanisms of the american grapes are still unknown. The few avaiable data has been obtained from samples cultivated in field, where the conditions are not fully under control. Our study is focused on the biosynthesis of stilbenes, firstly resveratrol (the main exponent) and viniferins (resveratrol oligomers), in response to biotic stress in cell cultures of several Vitis species. Stilbene content was compared in cell extracts obtained from Vitis spp. cell lines subjected to elicitation with MeJa, grown under continuous darkness or 16/8 h (light/dark) photoperiod. The main stilbenes involved in resistance against phytopathogenic fungi were identified and quantified by means of HPLC. Unexpectedly, the obtained results indicate that cultivars of V. vinifera have a higher stilbene content than the american grapes. For instance, V. vinifera cv. Malvasia, in response to MeJa, increases the production of total stilbenes from 2.58 to 15.58 mg/g DW while Vitis rupestris cv. du Lot increases from 2.80 to 8.97 mg/g DW. Moreover, cell lines grown under photoperiod accumulate higher levels of total stilbenes, both constitutively and in response to elicitation (e.g. Malvasia: from 8.69 to 37.81 mg/g DW)

POLY(LACTIC-CO-GLYCOLIC) ACID NANOPARTICLES UPTAKE BY VITIS VINIFERA AND GRAPEVINE-PATHOGENIC FUNGI

Poly(lactic-co-glycolic) acid (PLGA)-based NPs are currently considered among the most promising drug carriers, nevertheless their use in plants has never been investigated. In this work, for the first time, we demonstrated the ability of PLGA NPs to cross the plant cell wall and membrane of Vitis vinifera cell cultures and grapevine-pathogenic fungi. By means of fluorescence microscopy, we established that PLGA NPs can enter in grapevine leaf tissues through stomata openings and that they can be absorbed by the roots and transported to the shoot through vascular tissues. TEM analysis on cultured cells showed that NPs ≤ 50 nm could enter cells, while bigger ones remained attached to the cell wall. Viability tests demonstrated that PLGA NPs were not cytotoxic for V. vinifera-cultured cells. The cellular uptake of PLGA NPs by some important grapevine-pathogenic fungi has also been observed, thus suggesting that PLGA NPs could be used to deliver antifungal compounds within fungal cells. Overall the results reported suggest that such NPs may play a key role in future developments of agrobiotechnologies, as it is currently happening in biomedicine