Multitraits evaluation of a Solanum pennellii introgression tomato line challenged by combined abiotic stress.

Rising daily temperatures and water shortage are two of the major concerns in agriculture. In this work, we analysed the tolerance traits in a tomato line carrying a small region of the Solanum pennellii wild genome (IL12-4-SL) when grown under prolonged conditions of single and combined high temperature and water stress. When exposed to stress, IL12-4-SL showed higher heat tolerance than the cultivated line M82 at morphological, physiological, and biochemical levels. Moreover, under stress IL12-4-SL produced more flowers than M82, also characterized by higher pollen viability. In both lines, water stress negatively affected photosynthesis more than heat alone, whereas the combined stress did not further exacerbate the negative impacts of drought on this trait. Despite an observed decrease in carbon fixation, the quantum yield of PSII linear electron transport in IL12-4-SL was not affected by stress, thereby indicating that photochemical processes other than CO2 fixation acted to maintain the electron chain in oxidized state and prevent photodamage. The ability of IL12-4-SL to tolerate abiotic stress was also related to the intrinsic ability of this line to accumulate ascorbic acid. The data collected in this study clearly indicate improved tolerance to single and combined abiotic stress for IL12-4-SL, making this line a promising one for cultivation in a climate scenario characterized by frequent and long-lasting heatwaves and low rainfall.

The efficient physiological strategy of a novel tomato genotype to adapt to chronic combined water and heat stress.

Climate change is increasing the frequency of high temperature shocks and water shortages, pointing to the need to develop novel tolerant varieties and to understand the mechanisms employed to withstand combined abiotic stresses. Two tomato genotypes, a heat-tolerant Solanum lycopersicum accession (LA3120) and a novel genotype (E42), previously selected as a stable yielding genotype under high temperatures, were exposed to single and combined water and heat stress. Plant functional traits, pollen viability and physiological (leaf gas exchange and chlorophyll a fluorescence emission measurements) and biochemical (antioxidant content and antioxidant enzyme activity) measurements were carried out. A Reduced Representation Sequencing approach allowed exploration of the genetic variability of both genotypes to identify candidate genes that could regulate stress responses. Both abiotic stresses had a severe impact on plant growth parameters and on the reproductive phase of development. Growth parameters and leaf gas exchange measurements revealed that the two genotypes used different physiological strategies to overcome individual and combined stresses, with E42 having a more efficient capacity to utilize the limiting water resources. Activation of antioxidant defence mechanisms seemed to be critical for both genotypes to counteract combined abiotic stresses. Candidate genes were identified that could explain the different physiological responses to stress observed in E42 compared with LA3120. Results here obtained have shown how new tomato genetic resources can be a valuable source of traits for adaptation to combined abiotic stresses and should be used in breeding programmes to improve stress tolerance in commercial varieties.

HepG2 and human healthy hepatocyte in vitro culture and co-culture in PCL electrospun platforms.

The discovery of new drugs to treat pathological cells in the case of aggressive liver primary cancer is imposing the identification of high-throughput screening systems to predict the in vivo response of new therapeutic molecules, in order to reduce current use of animals and drug testing costs. Recently, micro/nanostructured scaffolds have been adopted to reproduce the hepatic microenvironment due to their higher similarity to the biological niche with respect to the traditional two-dimensional culture plate, so providing novel in vitro models for reliably understanding molecular mechanisms related to cancer cells activity. Herein, we propose the study of electrospun scaffolds made of polycaprolactone as in vitro model that can mimic the morphological organization of native extracellular matrix and the co-culture of hepatic cell lines-i.e., HepG2, human healthy hepatocytes (HHH). The micro- and nano-scale morphological features of fibers with diameter equal to (3.22 ± 0.42) µm and surface roughness of (17.84 ± 4.43) nm-allow the reproduction of the in vivo scenario influencing the adhesion and proliferation rate of the cultured cells. A much lower proliferation rate is observed for the HepG2 cells compared to the HHH cells, when cultured on the fibrous scaffolds over a time course of 4 weeks. Moreover, results on oxidative stress mechanisms indicate an antioxidant effect of fibers mainly in the case of co-colture, thus suggesting a promising use as new in vitro models to explore alternative therapeutic strategies in hepatocarcinoma treatment.

Navigator® and SmartPilot® View are helpful in guiding anesthesia and reducing anesthetic drug dosing.

BACKGROUND: The recently introduced Navigator® (GE Healthcare, Helsinki, Finland) and SmartPilot® View (Dräger Medical, Lübeck, Germany) show the concentrations and predicted effects of combined anesthetic drugs, and should facilitate more precisely their titration. Our aim was to evaluate if Navigator® or SmartPilot® View guided anesthesia was associated with a good quality of analgesia, depth of hypnosis and may reduce anesthetic requirements. METHODS: We performed a prospective non-randomized study. Sixty ASA I-II patients undergoing balanced general anesthesia for abdominal and plastic surgery were enrolled. Patients were divided in 4 groups. Group 1 (N. 15) and group 3 (N. 15) were cases in whom anesthesia was performed with standard monitoring plus the aid of Navigator® (Nav) or SmartPilot® View (SPV) display. Group 2 (N. 15) and group 4 (N. 15) were controls in whom anesthesia was performed with standard monitoring (heart rate, NIBP, SpO2, end-tidal CO2, end-expired sevoflurane concentration, train of four, Bispectral Index [Aspect Medical Systems, Natick, MA, USA] or Entropy [GE Healthcare]). Patients' vital parameters and end-expired sevoflurane concentration were recorded during anesthesia. RESULTS: All patients recovered uneventfully and showed hemodynamic stability. End-tidal sevoflurane concentrations values [median (min-max)], during maintenance of anesthesia, were significantly (P<0.05) lower in SPV [1.1% (0.8-1.5)] and Nav [1%(0.8-1.8)] groups compared to SPV-control group [1.5%(1-2.5)] and Nav-control group [1.5%(0.8-2)]. BIS and entropy values were respectively higher in the SPV group [53 (46-57)] compared to the control group [43 (37-51)] (P<0.05) and Nav group [53 (43-60)] compared to the control group [41 (35-51)] (P<0.05). No significant differences in Remifentanil dosing were observed in the four groups. CONCLUSION: Navigator® and SmartPilot® View may be of clinical use in monitoring adequacy of anesthesia. Both displays can optimize the administration and monitoring of anesthetic drugs during general anesthesia and may reduce the consumption of volatile anesthetic agents.

Femoral and sciatic nerves block (BiBlock) in orthopedic traumatologic lower limbs surgery in patients with multiple sclerosis.

Multiple sclerosis is a progressive demyelinating disease which affects large areas of the brain and of the spinal cord. Stressful events, surgical procedures, general anaesthesia and central blocks seem to be responsible for relapses, with worsening of the disease. So, when we scheduled 2 patients with multiple sclerosis for lower limbs orthopedic traumatologic surgery, we decided to use a peripheral block, and in particular a BiBlock. The patients' evaluation in the immediate postoperative course and 30 days after surgery has shown no relapses of the disease. In the literature, however, data about anaesthesia and multiple sclerosis are few and controversial, sometimes in contrast. Anyway, the use of peripheral blocks has neither anatomic, nor metabolic interferences with the lesion sites of multiple sclerosis. In conclusion, peripheral block is safe and it is the technique of choice for this type of patients, when surgery allows it.

Desflurane and sevoflurane in elderly patients during general anesthesia: a double blind comparison.

AIM: To investigate pulmonary wash-out of sevoflurane and desflurane and the quality of recovery from anesthesia in elderly patients. METHODS: Thirty-six patients aged >65 years, ASA II, were assigned in a double blind fashion to either desflurane (n=18) or sevoflurane (n=18) anesthesia. All received propofol 2 mg/kg and remifentanil 0.2 microg/kg/min for induction and 0.6 mg/kg of rocuronium. When the trachea was intubated volatile anaesthetic was administered. All data were recorded 1, 3, 5, 15, 30 min after intubation and then every 15 min. All data were recorded 1, 2, 3, 4, 5 min after suspension of all agents. Once extubated simple orders and questions were given every minute, times of appropriate response were noted. The patients were then transferred to the recovery room, until discharge to the floor. Postoperative pain control was obtained by a continuous iv infusion of ketorolac 60 mg and tramadol 100 mg. The latter was incremented by supplemental boluses of 50 mg according to patient needs (VAS <4) up to a maximum of 300 mg/24h. RESULTS: The F(A)/F(A0) ratio was lower in the desflurane group after halogenated agent suspension (p= or <0.05). Desflurane proved to have a faster wash out curve with respect to sevoflurane. Early recovery, as indicated by the time necessary to appropriately answer simple questions after the discontinuation of anesthetics, showed a significant advantage for desflurane (p= or <0.05). VAS was higher in the desflurane group as well as the needs for postoperative analgesia. CONCLUSIONS: Patients receiving desflurane reported faster recovery from anesthesia but an earlier and more intense perception of pain after surgery.

Peri-intubation cardiovascular response during low dose remifentanil or sufentanil administration in association with propofol TCI. A double blind comparison.

AIM: The aim of this study was to compare the effects on cardiovascular modifications induced by tracheal intubation when low dose infusion of remifentanil or sufentanil are used in association with propofol target controlled infusion. METHODS: Sixty normotensive, ASA I-II-III, Mallampati Score <3 undergoing general anaesthesia for major elective abdominal surgery, received i.m. midazolam (0.05 mg/kg) and atropine (0.01 mg/kg) 30 min before induction. They were randomly divided in a double-blind fashion into two groups receiving for induction respectively target controlled infusion of propofol (site-effect 3.0 microg/ml) and sufentanil (0.01 microg/kg/min) or remifentanil (0.1 microg/kg/ min) infusion. Rocuronium (0.6 mg/kg) was administered. Following intubation, the lungs were mechanically ventilated with an oxygen/air mixture. RESULTS: Bispectral index score and haemodynamic variables were recorded at baseline, after induction, during intubation and 1-3-5 min after the trachea was intubated. No differences in systolic and diastolic arterial pressure were observed in the sufentanil group, while the remifentanil group showed transient systolic and diastolic variations after intubating manoeuvers. The heart rate and bispectral index score were not affected in either group. CONCLUSION: In healthy normotensive patients the use of a small dose of either remifentanil or sufentanil after standard midazolam premedication, proved to be an effective strategy to blunt the cardiovascular response to intubation.

Anaestethic problems in Sanfilippo syndrome. A rare case of adult patient.

The authors report the case of a female patient (41 years old) affected by mucopolysaccharidosis type III or Sanfilippo syndrome submitted to a gynecologic surgical procedure and describe the main anesthesiologic problems. A sub-arachnoid anesthesia with hyperbaric Bupivacain 0.5% was used. This technique proved to be safe and convenient without peri- and postoperative complications.

Small doses of remifentanil and alfetanil in continuous total intravenous anesthesia in major abdominal surgery. A double blind comparison.

AIM: The purpose of this study was to test the safety and efficacy of small doses of remifentanil and alfentanil in a continuous total intravenous anesthesia technique for patients undergoing major abdominal surgery. METHODS: Sixty patients were enrolled in the study, and received in a double blind fashion either remifentanil (0.1 microg/kg/min) or alfentanil (alfentanil 0.75 microg/kg/min) in association with propofol (12 mg/kg/h at induction; 6-9 mg/kg/h for maintenance) and cisatracurium. Hemodynamic data, hypnosis monitoring data (Bispectral Index Score), ventilatory parameters and settings, drug utilisation were monitored during stress moments and during all the intraoperative period. Patients were evaluated also in the first 6 postoperative hours. RESULTS: Mean amount of propofol for induction (BIS<60) was lower in the remifentanil group than in the alfentanil group. Significantly fewer patients receiving remifentanil responded to intubation in comparison with patients receiving alfentanil in terms of non invasive blood pressure (>30 mmHg) and heart rate variations. Significantly more patients receiving alfentanil had 1 or more responses to surgery. Incidence of hypotension was significantly higher in patients receiving remifentanil. There were no differences between the 2 groups in the times for spontaneous respiration, adequate respiration, adequate responsivness (OAA/s=5) and discharge from the recovery room. Time to extubation resulted slightly shorter (p<0.05) in patients who received remifentanil. CONCLUSIONS: The use of remifentanil and alfentanil in association with propofol, in a continuous infusion total intravenous anesthesia technique, demonstrated to be safe and reliable strategies.

Enalapril: benefit-to-risk ratio in hypertensive patients.

Enalapril is an effective agent in the treatment of mild to severe hypertension. It is equally effective in elderly and young adult patients but appears to be more effective in white than in black hypertensive patients. Following treatment with enalapril, an assessment of maximum exercise performance found a decrease in total peripheral resistance without significant changes in cardiac output, heart rate, or stroke volume compared with pretreatment values. In addition, there have been reports of reversal of left ventricular hypertrophy in enalapril-treated hypertensive patients. Enalapril is also effective and well tolerated in hypertensive patients with renal impairment of varying etiology. The most common adverse experiences reported in controlled clinical trials were headache (5.2%), dizziness (4.3%), and fatigue (3.0%). In high-risk hypertensive patients, no enalapril-treated neutropenia, proteinuria, dysgeusia, or ageusia were reported. It may be concluded that the benefit-to-risk ratio of enalapril is among the best of the antihypertensive therapies currently available.

Lisinopril dose-response relationship in essential hypertension.

1. This was a multicentre, double-blind, parallel study in 216 patients with mild to moderate (supine diastolic blood pressure = 95-115 mm Hg) essential hypertension. 2. After a 4-week placebo washout, patients were randomized to placebo or lisinopril 1.25, 5.20 or 80 mg once daily for 6 consecutive weeks. Supine and erect blood pressure was measured 24 h postdose at the end of weeks -2, 0, 2, 4, and 6. 3. There was a linear dose-response relationship for both supine and erect blood pressure. Diastolic blood pressure reductions in the lisinopril 20 and 80 mg day-1 groups were significantly greater than in the placebo or lisinopril 1.25 and 5 mg day-1 groups. 4. Lisinopril, at doses up to 80 mg day-1, was well tolerated.

Sequence and structure of the yeast galactose transporter.

The previously cloned GAL2 gene of the Saccharomyces cerevisiae galactose transporter has been sequenced. The nucleotide sequence predicts a protein with 574 amino acids (Mr, 63,789). Hydropathy plots suggest that there are 12 membrane-spanning segments. The galactose transporter shows both sequence and structural homology with a superfamily of sugar transporters which includes the human HepG2-erythrocyte and fetal muscle glucose transporters, the rat brain and liver glucose transporters, the Escherichia coli xylose and arabinose permeases, and the S. cerevisiae glucose, maltose, and galactose transporters. Sequence and structural motifs at the N-terminal and C-terminal regions of the proteins support the view that the genes of this superfamily arose by duplication of a common ancestral gene. In addition to the sequence homology and the presence of the 12 membrane-spanning segments, the members of the superfamily show characteristic lengths and distributions of the charged, hydrophilic connecting loops. There is indirect evidence that the transporter is an N-glycoprotein. However, its only N-glycosylation site occurs in a charged, hydrophilic segment. This could mean that this segment is part of a hydrophilic channel in the membrane. The transporter has a substrate site for the cyclic AMP-dependent protein kinase which may be a target of catabolite inactivation. The transporter lacks a strong sequence enriched for proline (P), glutamate (E), aspartate, serine (S), and threonine (T) and flanked by basic amino acids (PEST sequence) even though it has a short half-life. Mechanisms for converting the poor PEST to a possible PEST sequence are considered. Like the other members of the superfamily, the galactose transporter lacks a signal sequence.

Characteristics of galactose transport in Saccharomyces cerevisiae cells and reconstituted lipid vesicles.

Growth on galactose induces two transport processes, a high-affinity and a low-affinity process. The most important results of a comparison of the two processes were that (i) both depended on GAL2 expression, (ii) only the high-affinity process required galactokinase, (iii) both were down-regulated by catabolite inactivation, (iv) neither was significantly inhibited by carbonyl cyanide-p-trifluoromethoxy-phenyl-hydrazone, (v) neither was differentially inhibited by silver nitrate or mercuric chloride, and (vi) transport activity with a Km closer to that of the low-affinity process of whole cells was reconstituted in fused phospholipid membrane vesicles.

Role of cyclic-AMP-dependent protein kinase in catabolite inactivation of the glucose and galactose transporters in Saccharomyces cerevisiae.

The derepressed high-affinity glucose transport system and the induced galactose transport system are catabolite inactivated when cells with these transport systems are incubated with glucose. The role of the cyclic AMP cascade in the catabolite inactivation of these transport systems was shown by using mutants affected in the activity of cyclic-AMP-dependent protein kinase (cAPK). In tpk1(w) mutants with reduced cAPK activity, the sugar transport systems were expressed but were not catabolite inactivated. In bcy1 mutants with unbridled cAPK activity resulting from a defective regulatory subunit, the transport systems were absent or present at low levels.

Relationship between low- and high-affinity glucose transport systems of Saccharomyces cerevisiae.

The high-affinity glucose transport process in Saccharomyces cerevisiae whole cells was regulated by catabolite repression and inactivation. The low-affinity process was constitutive, and its activity was inhibited in proportion to the extent of derepression of the high-affinity process. The latter finding suggests that there is some regulatory relationship between the two processes.