Testing the IMEter on rice introns and other aspects of intron-mediated enhancement of gene expression

In many eukaryotes, spliceosomal introns are able to influence the level and site of gene expression. The mechanism of this Intron Mediated Enhancement (IME) has not yet been elucidated, but regulation of gene expression is likely to occur at several steps during and after transcription. Different introns have different intrinsic enhancing properties, but the determinants of these differences remain unknown. Recently, an algorithm called IMEter, which is able to predict the IME potential of introns without direct testing, has been proposed. A computer program was developed for Arabidopsis thaliana and rice (Oryza sativa L.), but was only tested experimentally in Arabidopsis by measuring the enhancement effect on GUS expression of different introns inserted within otherwise identical plasmids. To test the IMEter potential in rice, a vector bearing the upstream regulatory sequence of a rice β-tubulin gene (OsTub6) fused to the GUS reporter gene was used. The enhancing intron interrupting the OsTub6 5′-UTR was precisely replaced by seven other introns carrying different features. GUS expression level in transiently transformed rice calli does not significantly correlate with the calculated IMEter score. It was also found that enhanced GUS expression was mainly due to a strong increase in the mRNA steady-state level and that mutations at the splice recognition sites almost completely abolished the enhancing effect. Splicing also appeared to be required for IME in Arabidopsis cell cultures, where failure of the OsTub6 5′ region to drive high level gene expression could be rescued by replacing the poorly spliced rice intron with one from Arabidopsis

Idiopathic acute pancreatitis: a single-center investigation of clinical and biochemical features

Acute pancreatitis (AP) is termed as idiopathic (IAP) when the underlying conditions of pancreatic inflammation remain unknown. The aim of this study was to identify different clinical features in patients with IAP and AP of known aetiology. All patients hospitalized in our Gastroenterology Unit with an initial diagnosis of AP were recruited. AP was classified as of known aetiology or idiopathic according to clinical examination, serum biochemistry testing, and radiological imaging investigations, and clinical data in both patient groups were compared. A total of 127 patients (80 males, mean age: 57 years) were eligible for the analysis, 92 of which (73%) with AP of known aetiology and 35 (27%) with IAP. The major causes of AP were biliary obstruction (65%) or alcohol abuse (25%). Previous cholecystectomy was more frequent in patients with AP of known aetiology than in patients with IAP (14% versus 0%); patients with IAP showed lower gamma-glutamyl transpeptidase levels, lower daily alcohol intake, and higher frequency of gastroenteritis than patients with AP of known aetiology (34.3% versus 15.2%). Previous intake of nonsteroidal anti-inflammatory drugs was more frequent in patients with IAP than in patients with AP of known aetiology (23% versus 0%). No further differences in clinical features were found between the two patient groups. IAP accounts for almost 20% of cases of AP. An association of AP with gastroenteritis or the use of NSAIDs should be considered if time-related with disease onset, especially in patients with no recurrent attacks

Serum Levels of Granulocyte-Macrophage-colony-stimulating Factor and Stem-cell Factor During Liver Regeneration after Partial Hepatectomy in Humans

Background: Multiple biological functions have been recognized regarding Granulocyte Macrophage-Colony Stimulating Factor (GM-CSF) and Stem Cell Factor (SCF).Aim: To evaluate the serum changes of GM-CSF and SCF in patients undergoing surgical resection for liver tumor, in the regenerative phase after surgery in order to identify the possible relationship with the patient, tumor or surgical variables.Methods: Thirty-two consecutive patients (50% male, median age 66), undergoing hepatic resection of liver neoplasm, were evaluated. The liver tumor was Hepatocellular Carcinoma (HCC) in 44% of cases. Other tumors were cholangiocarcinoma and metastasis. Serum levels of GM-CSF and SCF were assessed at baseline and 2 days, 7 days and 4 weeks after surgery. Personal and clinical patient data were also recorded. The statistical analysis was carried out using t-test for unpaired data or ANOVA (repeated measure) for continuous variables and Fisher test for discrete variables.Results: GM-CSF levels remained constant after surgery and were compared to baseline values. SCF levels, on the other hand, increased during the time, after surgery. The evaluation of SCF levels (fold increase) according to surgical, patient and tumor variables evidenced some differences. At day 7 and week 4, SCF levels were statistically increased: i) in patients undergoing a large resection in comparison with others (p<0.05); ii) in patients non-cirrhotic in comparison with cirrhotic ones (p=0.02) and finally; iii) in patients with non-HCC tumor in comparison with HCC ones (p=0.02).Conclusion: During liver regeneration in humans, SCF serum levels are increased allowing to hypothesize a possible role of this chemokine during tissue growth and remodeling

miRNA-23a modulates sodium-hydrogen exchanger 1 expression: studies in medullary thick ascending limb of salt-induced hypertensive rats

Background. The kidney is the main organ in the pathophys- iology of essential hypertension. Although most bicarbonate reabsorption occurs in the proximal tubule, the medullary thick ascending limb (mTAL) of the nephron also maintains acid–base balance by contributing to 25% of bicarbonate reab- sorption. A crucial element in this regulation is the sodium- hydrogen exchanger 1 (NHE1), a ubiquitous membrane protein controlling intracellular pH, where proton extrusion is driven by the inward sodium ux. MicroRNA (miRNA) expression of hypertensive patients signi cantly di ers from that of normotensive subjects. The aim of this study was to determine the functional role of miRNA alterations at the mTAL level. Methods. By miRNA microarray analysis, we identi ed miRNA expression pro les in isolated mTALs from high sodium intake–induced hypertensive rats (HSD) versus their normotensive counterparts (NSD). In vitro validation was carried out in rat mTAL cells. Results. Five miRNAs involved in the onset of salt-sensitive hypertension were identi ed, including miR-23a, which was bioinformatically predicted to target NHE1 mRNA. Data demonstrated that miRNA-23a is downregulated in the mTAL of HSD rats while NHE1 is upregulated. Consistently, transfec- tion of an miRNA-23a mimic in an mTAL cell line, using a viral vector, resulted in NHE1 downregulation. Conclusion. NHE1, a protein involved in sodium reabsorption at the mTAL level and blood pressure regulation, is upregulated in our model. This was due to a downregulation of miRNA-23a. Expression levels of this miRNA are in uenced by high sodium intake in the mTALs of rats. The downregulation of miRNA- 23a in humans a ected by essential hypertension corroborate our data and point to the potential role of miRNA-23a in the regulation of mTAL function following high salt intake

L’ Assemblea costituente italiana nell’opinione pubblica europea

Il volume è improntato alla volontà di sprovincializzare il dibattito sulla Costituzione italiana attraverso l'analisi delle reazioni dell'opinione pubblica europea di fronte alle vicende politico istituzionali del nostro dopoguerra.Tale contributo si colloca in un momento politico di difficoltà per la Repubblica nata dalla Resistenza e fondata sulla Costituzione,segnato dalla crisi dei partiti e da un dibattito che spinge verso una revisione costituzionale

A prospective non-randomized controlled, multicenter trial comparing Appendectomy and Conservative Treatment for Patients with Uncomplicated Acute Appendicitis (the ACTUAA study)

Purpose: Acute appendicitis (AA) is among the most common causes of lower abdominal pain and admissions to the emergency department. Over the past 20 years, there has been a renewed interest in the conservative management of uncomplicated AA, and several studies demonstrated that an antibiotic-first strategy is a viable treatment option for uncomplicated AA. The aim of this prospective non-randomized controlled, multicenter trial is to compare antibiotic therapy and emergency appendectomy as treatment for patients with uncomplicated AA confirmed by US and/or CT or MRI scan. Methods: All adult patients in the age range 18 to 65 years with suspected AA, consecutively admitted to the Surgical Department of the 13 participating Italian Hospitals, will be invited to take part in the study. A multicenter prospective collected registry developed by surgeons, radiologists, and pathologists with expertise in the diagnosis and treatment of uncomplicated acute appendicitis represents the best research method to assess the long-term role of antibiotics in the management of the disease. Comparison will be made between surgical and antibiotic-first approaches to uncomplicated AA through the analysis of the primary outcome measure of complication-free treatment success rate based on 1-year follow-up. Quality of life, length of hospital stay, pain evaluation, and time to return to normal activity will be evaluated as secondary outcome measures. Trial registration: Clinicaltrials.gov ID: NCT03080103