Optimization of transgene action at the post-transcriptional level: high quality parthenocarpic fruits in industrial tomatoes

Background: Genetic engineering of parthenocarpy confers to horticultural plants the ability toproduce fruits under environmental conditions that curtail fruit productivity and quality. TheDefH9-iaaM transgene, whose predicted action is to confer auxin synthesis specifically in theplacenta, ovules and derived tissues, has been shown to confer parthenocarpy to several plantspecies (tobacco, eggplant, tomato) and varieties.Results: UC82 tomato plants, a typical cultivar used by the processing industry, transgenic for theDefH9-iaaM gene produce parthenocarpic fruits that are malformed. UC82 plants transgenic forthe DefH9-RI-iaaM, a DefH9-iaaM derivative gene modified in its 5'ULR by replacing 53 nucleotidesimmediately upstream of the AUG initiation codon with an 87 nucleotides-long sequence derivedfrom the rolA intron sequence, produce parthenocarpic fruits of high quality. In an in vitro translationsystem, the iaaM mRNA, modified in its 5'ULR is translated 3\u20134 times less efficiently than theoriginal transcript. An optimal expressivity of parthenocarpy correlates with a reduced transgenemRNA steady state level in DefH9-RI-iaaM flower buds in comparison to DefH9-iaaM flower buds.Consistent with the known function of the iaaM gene, flower buds transgenic for the DefH9-RI-iaaMgene contain ten times more IAA than control untransformed flower buds, but five times less thanDefH9-iaaM flower buds.Conclusions:: By using an auxin biosynthesis transgene downregulated at the post-transcriptionallevel, an optimal expressivity of parthenocarpy has been achieved in a genetic background notsuitable for the original transgene. Thus, the method allows the generation of a wider range ofexpressivity of the desired trait in transgenic plants

Improved Drought Stress Response in Alfalfa Plants Nodulated by an IAA Over-producing Rhizobium Strain

The drought\u2013stress response in plant involves the cross-talk between abscisic acid (ABA) and other phytohormones, such as jasmonates and ethylene. The auxin indole-3- acetic acid (IAA) plays an integral part in plant adaptation to drought stress. Investigation was made to see how the main auxin IAA interacted with other plant hormones under water stress, applied through two different growth conditions (solid and hydroponic). Medicago sativa plants nodulated by the Ensifer meliloti wild type 1021 (Ms-1021) and its IAA-overproducing RD64 derivative strains (Ms-RD64) were subjected to drought stress, comparing their response. When the expression of nifH gene and the activity of the nitrogenase enzyme were measured after stress treatments, Ms-RD64 plants recorded a significantly weaker damage. These results were correlated with a lower biomass reduction, and a higher Rubisco protein level measured for the Ms-RD64- stressed plants as compared to the Ms-1021-stressed ones. It has been verified that the stress response observed for Ms-RD64-stressed plants was related to the production of greater amount of low-molecular-weight osmolytes, such as proline and pinitol, measured in these plants. For the Ms-RD64 plants the immunoblotting analysis of thylakoid membrane proteins showed that some of the photosystem proteins increased after the stress. An increased non-photochemical quenching after the stress was also observed for these plants. The reduced wilting signs observed for these plants were also connected to the significant down-regulation of the MtAA03 gene involved in the ABA biosynthesis, and with the unchanged expression of the two genes (Mt- 2g006330 and Mt-8g095330) of ABA signaling. When the expression level of the ethylene-signaling genes was evaluated by qPCR analysis no significant alteration of the key positive regulators was recorded for Ms-RD64-stressed plants. Coherently, these plants accumulated 40% less ethylene as compared to Ms-1021-stressed ones. The results presented herein indicate that the variations in endogenous IAA levels, triggered by the overproduction of rhizobial IAA inside root nodules, positively affected drought stress response in nodulated alfalfa plants

An Investigation of Nurses' Knowledge, Attitudes, and Practices Regarding Disinfection Procedures in Italy

<p>Abstract</p> <p>Background</p> <p>This study assessed the level of knowledge, attitudes, and practice regarding disinfection procedures among nurses in Italian hospitals.</p> <p>Methods</p> <p>A face-to-face interview gathered the following information: demographic and practice characteristics; knowledge about the healthcare-associated infections (HAIs) and the disinfection practices; attitudes towards the utility of guidelines/protocols and perception of the risks of acquiring/transmitting HAIs; compliance with antisepsis/disinfection procedures; and sources of information.</p> <p>Results</p> <p>Only 29% acknowledged that urinary and respiratory tract infections were the two most common HAIs and this knowledge was significantly higher in those with a higher level of education. Attitudes towards the utility of guidelines/protocols for disinfection procedures showed a mean score of 9.1. The results of the linear regression model indicated a more positive attitude in female nurses, in those with a lower number of years of activity, and in those needing additional information about disinfection procedures. Nurses with higher educational level and with a higher perception of risk of transmitting an infectious disease while working were more likely to perform appropriate antisepsis of the surgical wound and handwashing before and after medication.</p> <p>Conclusions</p> <p>Plan of successful prevention activities about HAIs and provide pointers to help optimize disinfection procedures and infection prophylaxis and management are needed.</p

Epidemiology, practice of ventilation and outcome for patients at increased risk of postoperative pulmonary complications

BACKGROUND Limited information exists about the epidemiology and outcome of surgical patients at increased risk of postoperative pulmonary complications (PPCs), and how intraoperative ventilation was managed in these patients. OBJECTIVES To determine the incidence of surgical patients at increased risk of PPCs, and to compare the intraoperative ventilation management and postoperative outcomes with patients at low risk of PPCs. DESIGN This was a prospective international 1-week observational study using the ‘Assess Respiratory Risk in Surgical Patients in Catalonia risk score’ (ARISCAT score) for PPC for risk stratification. PATIENTS AND SETTING Adult patients requiring intraoperative ventilation during general anaesthesia for surgery in 146 hospitals across 29 countries. MAIN OUTCOME MEASURES The primary outcome was the incidence of patients at increased risk of PPCs based on the ARISCAT score. Secondary outcomes included intraoperative ventilatory management and clinical outcomes. RESULTS A total of 9864 patients fulfilled the inclusion criteria. The incidence of patients at increased risk was 28.4%. The most frequently chosen tidal volume (VT) size was 500 ml, or 7 to 9 ml kg1 predicted body weight, slightly lower in patients at increased risk of PPCs. Levels of positive end-expiratory pressure (PEEP) were slightly higher in patients at increased risk of PPCs, with 14.3% receiving more than 5 cmH2O PEEP compared with 7.6% in patients at low risk of PPCs (P < 0.001). Patients with a predicted preoperative increased risk of PPCs developed PPCs more frequently: 19 versus 7%, relative risk (RR) 3.16 (95% confidence interval 2.76 to 3.61), P < 0.001) and had longer hospital stays. The only ventilatory factor associated with the occurrence of PPCs was the peak pressure. CONCLUSION The incidence of patients with a predicted increased risk of PPCs is high. A large proportion of patients receive high VT and low PEEP levels. PPCs occur frequently in patients at increased risk, with worse clinical outcome

Bacteria-inducing legume nodules involved in the improvement of plant growth, health and nutrition

Bacteria-inducing legume nodules are known as rhizobia and belong to the class Alphaproteobacteria and Betaproteobacteria. They promote the growth and nutrition of their respective legume hosts through atmospheric nitrogen fixation which takes place in the nodules induced in their roots or stems. In addition, rhizobia have other plant growth-promoting mechanisms, mainly solubilization of phosphate and production of indoleacetic acid, ACC deaminase and siderophores. Some of these mechanisms have been reported for strains of rhizobia which are also able to promote the growth of several nonlegumes, such as cereals, oilseeds and vegetables. Less studied are the mechanisms that have the rhizobia to promote the plant health; however, these bacteria are able to exert biocontrol of some phytopathogens and to induce the plant resistance. In this chapter, we revised the available data about the ability of the legume nodule-inducing bacteria for improving the plant growth, health and nutrition of both legumes and nonlegumes. These data showed that rhizobia meet all the requirements of sustainable agriculture to be used as bio-inoculants allowing the total or partial replacement of chemicals used for fertilization or protection of crops

Epidemiology, practice of ventilation and outcome for patients at increased risk of postoperative pulmonary complications: LAS VEGAS - An observational study in 29 countries

BACKGROUND Limited information exists about the epidemiology and outcome of surgical patients at increased risk of postoperative pulmonary complications (PPCs), and how intraoperative ventilation was managed in these patients. OBJECTIVES To determine the incidence of surgical patients at increased risk of PPCs, and to compare the intraoperative ventilation management and postoperative outcomes with patients at low risk of PPCs. DESIGN This was a prospective international 1-week observational study using the ‘Assess Respiratory Risk in Surgical Patients in Catalonia risk score’ (ARISCAT score) for PPC for risk stratification. PATIENTS AND SETTING Adult patients requiring intraoperative ventilation during general anaesthesia for surgery in 146 hospitals across 29 countries. MAIN OUTCOME MEASURES The primary outcome was the incidence of patients at increased risk of PPCs based on the ARISCAT score. Secondary outcomes included intraoperative ventilatory management and clinical outcomes. RESULTS A total of 9864 patients fulfilled the inclusion criteria. The incidence of patients at increased risk was 28.4%. The most frequently chosen tidal volume (V T) size was 500 ml, or 7 to 9 ml kg−1 predicted body weight, slightly lower in patients at increased risk of PPCs. Levels of positive end-expiratory pressure (PEEP) were slightly higher in patients at increased risk of PPCs, with 14.3% receiving more than 5 cmH2O PEEP compared with 7.6% in patients at low risk of PPCs (P ˂ 0.001). Patients with a predicted preoperative increased risk of PPCs developed PPCs more frequently: 19 versus 7%, relative risk (RR) 3.16 (95% confidence interval 2.76 to 3.61), P ˂ 0.001) and had longer hospital stays. The only ventilatory factor associated with the occurrence of PPCs was the peak pressure. CONCLUSION The incidence of patients with a predicted increased risk of PPCs is high. A large proportion of patients receive high V T and low PEEP levels. PPCs occur frequently in patients at increased risk, with worse clinical outcome.</p

Studio sulla relazione tra la resistenza alla kanamicina ed il gene “Putative aminoglycoside 3’-phosphotranferase (PAPH) in Sinorhizobium meliloti ceppo 1021

Il ceppo selvatico di Sinorhizobium meliloti 1021 è sensibile alla kanamicina (30 mg/l) anche se Capela et al. (Proc. Natl. Acad. Sci. U.S.A.(2001), 98: 9877-9882), analizzando l’intero genoma, hanno trovato un gene codificante per una “putative aminoglycoside 3'-phosphotransferase” (PAPH), coinvolta nella resistenza ad antibiotici aminoglicosidici quali la kanamicina. Un mutante spontaneo (GM42) kanamicina resistente (200 mg/l) di Sinorhizobium meliloti 1021 è stato caratterizzato da un punto di vista molecolare e fenotipico per analizzare l’attività del gene PAPH. Il ceppo parentale e GM42 sono stati caratterizzati da un punto di vista microbiologico tramite il MicroLog System Biolog. I risultati mostrano pattern differenti tra il ceppo parentale ed il mutante sulla base della loro capacità di utilizzare o meno fonti di carbonio quali mannitolo o maltosio. L'amplificazione del gene PAPH tramite PCR ha prodotto un'unica banda sia nel ceppo wild-type 1021 che in GM42 e dal loro sequenziamento non è emersa alcuna differenza nucleotidica. Tuttavia, l’analisi trascrizionale mediante RT-PCR del gene PAPH ha evidenziato differenze tra i due ceppi. Infatti il wild-type non ha prodotto alcun amplicone, mentre GM42 esprime il gene PAPH in diverse condizioni di crescita saggiate. Sono state condotte ulteriori analisi mediante Western blotting sulle proteine totali del parentale e del ceppo mutante utilizzando l’anticorpo anti-neomicina fosfotransferasi II. GM42 contiene un peptide che immunoreagisce con l’anticorpo utilizzato, tale reattività è assente nel ceppo selvatico. Inoltre tale gene è stato clonato nel vettore di espressione pKK223-3 in E. coli TOP10 e i cloni ottenuti hanno acquisito resistenza alla kanamicina. Dal confronto dell’espressione genica effettuata mediante DNA-microarrays si evidenzia un’overespressione del gene PAPH e della regione a monte di tale gene nel mutante kanamicina resistente. Saranno condotte ulteriori indagini per la comprensione dei meccanismi genetici alla base del fenotipo del ceppo mutante GM42. Ciò rappresenterà un considerevole passo in avanti nello studio dei fattori di resistenza delle popolazioni naturali di S. meliloti 1021, aprendo la strada ad un razionale impiego in agricoltura sostenibile