Impact of ubiquitous inhibitors on the GUS gene reporter system: evidence from the model plants Arabidopsis, tobacco and rice and correction methods for quantitative assays of transgenic and endogenous GUS

<p>Abstract</p> <p>Background</p> <p>The β-glucuronidase (GUS) gene reporter system is one of the most effective and employed techniques in the study of gene regulation in plant molecular biology. Improving protocols for GUS assays have rendered the original method described by Jefferson amenable to various requirements and conditions, but the serious limitation caused by inhibitors of the enzyme activity in plant tissues has thus far been underestimated.</p> <p>Results</p> <p>We report that inhibitors of GUS activity are ubiquitous in organ tissues of Arabidopsis, tobacco and rice, and significantly bias quantitative assessment of GUS activity in plant transformation experiments. Combined with previous literature reports on non-model species, our findings suggest that inhibitors may be common components of plant cells, with variable affinity towards the <it>E. coli </it>enzyme. The reduced inhibitory capacity towards the plant endogenous GUS discredits the hypothesis of a regulatory role of these compounds in plant cells, and their effect on the bacterial enzyme is better interpreted as a side effect due to their interaction with GUS during the assay. This is likely to have a bearing also on histochemical analyses, leading to inaccurate evaluations of GUS expression.</p> <p>Conclusions</p> <p>In order to achieve reliable results, inhibitor activity should be routinely tested during quantitative GUS assays. Two separate methods to correct the measured activity of the transgenic and endogenous GUS are presented.</p

Stylar peroxidases and heteromorphic incompatibility reactions in Primula acaulis Hill («thrum» morph)

SUMMARYIn non-, self- and cross-pollinated styles of Primula acaulis «thrum» morph, the distribution of apoplastic peroxidase activity has been investigated, by means of the cytochemical treatment with diaminobenzidine. Apoplastic peroxidase activity was detected throughout the transmitting tract (at the «neck» region level) in non- and self-pollinated styles. The enzyme has been related to the predisposition of incompatible pollen tube rejection, no appreciable differences in enzyme distribution having been found between non- and self-pollinated styles since, in this morph, incompatible pollen tubes hardly ever reach the stylar neck. Compatible intermorph pollination caused the disappearance of apoplastic peroxidase activity in the central portion of the transmitting tract, where pollen tubes elongated, utilizing the reserves of the stylar cells, which appeared highly degenerate. Stylar apoplastic peroxidases seem then to play a role in heteromorphic incompatibility responses

Prevalence of hypospadias in Italy according to severity, gestational age and birthweight: an epidemiological study

<p>Abstract</p> <p>Background</p> <p>Hypospadias is a congenital displacement of the urethral meatus in male newborns, being either an isolated defect at birth or a sign of sexual development disorders. The aim of this study was to assess the prevalence rate of hypospadias in different Districts of Italy, in order to make a comparison with other countries all over the world.</p> <p>Methods</p> <p>We reviewed all the newborns file records (years 2001–2004) in 15 Italian Hospitals.</p> <p>Results</p> <p>We found an overall hypospadias prevalence rate of 3.066 ± 0.99 per 1000 live births (82.48% mild hypospadias, 17.52% moderate-severe). In newborns Small for Gestational Age (birthweight < 10^thpercentile) of any gestational age the prevalence rate of hypospadias was 6.25 per 1000 live births. Performing multivariate logistic regression analysis for different degrees of hypospadias according to severity, being born SGA remained the only risk factor for moderate-severe hypospadias (p = 0.00898) but not for mild forms (p > 0.1).</p> <p>Conclusion</p> <p>In our sample the prevalence of hypospadias results as high as reported in previous European and American studies (3–4 per 1000 live births). Pathogenesis of isolated hypospadias is multifactorial (genetic, endocrine and environmental factors): however, the prevalence rate of hypospadias is higher in infants born small for gestational age than in newborns with normal birth weight.</p

Double vs single internal thoracic artery harvesting in diabetic patients: role in perioperative infection rate

Background: The aim of this prospective study is to evaluate the role in the onset of surgical site infections of bilateral internal thoracic arteries harvesting in patients with decompensated preoperative glycemia. Methods: 81 consecutive patients with uncontrolled diabetes mellitus underwent elective CABG harvesting single or double internal thoracic arteries. Single left ITA was harvested in 41 patients (Group 1, 50.6%), BITAs were harvested in 40 (Group 2, 49.4%). The major clinical end points analyzed in this study were infection rate, type of infection, duration of infection, infection relapse rate and total hospital length of stay. Results: Five patients developed sternal SSI in the perioperative period, 2 in group 1 and 3 in group 2 without significant difference. All sternal SSIs were superficial with no sternal dehiscence. The development of infection from the time of surgery took 18.5 ± 2.1 and 7.3 ± 3.0 days for Groups 1 and 2 respectively. The infections were treated with wound irrigation and debridement, and with VAC therapy as well as with antibiotics. The VAC system was removed after a mean of 12.8 ± 5.1 days, when sterilization was achieved. The overall survival estimate at 1 year was 98.7%. Only BMI was a significant predictor of SSI using multivariate stepwise logistic regression analysis (Odds Ratio: 1.34; 95%Conficdence Interval: 1.02–1.83; p value: 0.04). In the model, the use of BITA was not an independent predictor of SSI. Conclusion: CABG with bilateral pedicled ITAs grafting could be performed safely even in diabetics with poor preoperative glycaemic control

Studi sia in vivo che in vitro sull'internalizzazione di proteine nei tubetti pollinici di diverse specie di Nicotiana in relazione ai processi di incompatibilit\ue0 gametofitica e incongruit\ue0 unilaterale interspecifica

Scopo di questo progetto di ricerca \ue8 studiare la penetrazione delle S-RNasi nei tubetti pollinici \u201cin vivo\u201d e \u201cin vitro\u201d, sia in relazione ai processi di auto-incompatibilit\ue0 (GSI) e di inconguit\ue0 unilaterale interspecifica (UR) che in relazione ai processi di internalizzazione