Characterization of Neuroendocrine Tumors of the Pancreas by Real-Time Quantitative Polymerase Chain Reaction. A Methodological Approach

The aim of this study was to assess the suitability of using real-time quantitative PCR (RT-qPCR) to characterize neuroendocrine (NE) tumors of the pancreas. For a series of tumors, we evaluated several genes of interest, and the data were matched with the “classical” immunohistochemical (IHC) features. In 21 cases, we extracted RNA from formalin-fixed paraffin-embedded (FFPE) blocks, and in nine cases, we also extracted RNA from fresh-frozen tissue. The RT-qPCR procedure was performed using two sets of customized arrays. The test using the first set, covering 96 genes of interest, was focused on assessing the feasibility of the procedure, and the results were used to select 18 genes indicative of NE differentiation, clinical behavior, and therapeutic responsiveness for use in the second set of arrays. Threshold cycle (Ct) values were used to calculate the fold-changes in gene expression using the 2-??Ct method. Statistical procedures were used to analyze the results, which were matched with the IHC and follow-up data. Material from fresh-frozen samples performed better in terms of the level of amplification, but acceptable and concordant results were also obtained from FFPE samples. In addition, high concordance was observed between the mRNA and protein expression levels of somatostatin receptor type 2A (R?=?0.52, p?=?0.016). Genes associated with NE differentiation, as well as the gastrin-releasing peptide receptor and O-6-methylguanine-DNA methyltransferase genes, were underexpressed, whereas angiogenesis-associated markers (CDH13 and SLIT2) were overexpressed in tissues with malignant behavior. The RT-qPCR procedure is practical and feasible in economic terms for the characterization of NE tumors of the pancreas and can complement morphological and IHC-based evaluations. Thus, the results of the RT-qPCR procedure might offer an objective basis for therapeutic choices.     

Clinical characteristics and outcome of diabetic patients with acute myocardial infarction. Data from the BLITZ-1 study.

BACKGROUND: The determinants of a worse outcome in diabetic patients after an acute myocardial infarction (AMI) are controversial. They include delayed hospital admission, worse clinical presentation and lesser efficacy of accepted therapeutic interventions. Therefore, to improve our knowledge, we aimed to describe the clinical characteristics, treatment options and short-term outcomes of diabetic patients in a survey of consecutive AMI subjects admitted to the Italian coronary care unit (CCU) network in the current era of reperfusion. METHODS: The BLITZ study prospectively enrolled patients with AMI, within 48 hours of symptom onset, admitted to 296 out of the 341 existing Italian CCUs from October 15 to 29, 2001. Diabetic status was recorded by collecting clinical history. In-hospital and post-discharge management and outcomes were collected up to 30 days from admission. RESULTS: Overall, 434 of 1959 enrolled patients (22%) had a clinical diagnosis of diabetes. Diabetic patients were older, more frequently women, had a worse coronary risk profile, and an unfavorable clinical presentation compared to non-diabetics. Among 1275 patients with ST-elevation AMI, diabetics (20%) received a similar proportion of any reperfusion therapy (61 vs 66%, p = 0.10), but significantly less primary percutaneous coronary angioplasty (9 vs 16%, p = 0.003). Diabetic patients were treated less often with oral beta-blockers than non-diabetics both during hospitalization (56 vs 64%, p = 0.003) and at discharge (54 vs 61%, p = 0.01). In contrast, in-hospital use of angiotensin-converting enzyme inhibitors (76 vs 67%, p = 0.0003), digitalis (10 vs 5%, p = 0.0005), and diuretics (54 vs 36%, p < 0.0001) was more frequent among diabetics. During their index admission, subjects with diabetes had higher in-hospital mortality (11 vs 6%, p = 0.0004), as well as higher rates of reinfarction (6 vs 2%, p = 0.0003), new congestive heart failure (28 vs 14%, p < 0.0001), cardiogenic shock (10 vs 5%, p = 0.0005) or recurrent angina (22 vs 16%, p = 0.0034). A similar pattern was observed at 30-day follow-up. At multivariate analysis, diabetic status was not confirmed to be an independent predictor of 30-day mortality. CONCLUSIONS: Although diabetic patients with AMI admitted to the Italian CCU network have a higher in-hospital and 30-day morbidity and mortality rates compared to non-diabetics, a clinical diagnosis of diabetes has no independent predictive value on short-term outcome.     

Saccharomyces boulardii in Maintenance Treatment of Crohn’s Disease

The possible role of Saccharomyces boulardii, a nonpathogenic yeast with beneficial effects on the human intestine, in the maintenance treatment of Crohns disease has been evaluated. Thirty-two patients with Crohns disease in clinical remission (CDAI < 150) were randomly treated for six months with either mesalamine 1 g three times a day or mesalamine 1 g two times a day plus a preparation of Saccharomyces boulardii 1 g daily. Clinical relapses as assessed by CDAI values were observed in 37.5% of patients receiving mesalamine alone and in 6.25% of patients in the group treated with mesalamine plus the probiotic agent. Our results suggest that Saccharomyces boulardii may represent a useful tool in the maintenance treatment of Crohns disease. However, in view of the products cost, further controlled studies are needed to confirm these preliminary data.     

Biphasic Diurnal Periodicity in Bleeding from Peptic Ulcer

Objective : To evaluate if tbere was periodicity in the manifestations of gastrointestinal bleeding (hemateme-sis and melena). Method : This is a multicenter prospective study carried out in the Endoscopy Units of eight hospitals. At the time of the emergency endoscopy, the following data were collected: age, sex, endoscopic diagnosis, solar hour of the first hematemesis (vomiting of bright red or tarry black material) and of the first melena (black or bloody soft stools), and any drugs taken during the week before the bleeding episode, regardless of the dose. Results : 806 patients were studied. Bleeding was from peptic ulcer in 405 patients (50%), from esophageal varices in 197 (24%), and from other sources in the remainder. Analysis using single cosinor statistics showed a nonrandom distribution in bleeding from peptic ulcer, whether presenting first with hematemesis (p = 0.02) or melena (p = 0.03). There were two peaks at 6:45 AM and 6:45 PM for hemate-mesis and at 7:25 AM and 7:25 PM for melena, representing a biphasic diurnal (ultradian) rhythm. Conclusions : This study shows that bleeding due to peptic ulcer has a biphasic diurnal periodicity. 1 his has potential importance for the pathogenesis of bleeding, for the management of gastrointestinal hemorrhage and the administration of drugs known to cause peptic ulcer bleeding.     

Human immunodeficiency virus type 1 and other viral co-infections among young heterosexual men and women in Argentina

Infections with hepatitis C virus, (HCV), hepatitis B virus (HBV), and human T lymphotropic type I/II (HTLV-I/II) virus are commonly found in patients infected with human immunodeficiency virus type 1 (HIV-1). We conducted a seroepidemiologic study among 174 HIV-positive heterosexuals in Buenos Aires, Argentina in 1999. Evidence of exposure to HCV, HBV, and HTLV-I/II was found in 32%, 17%, and 5%, respectively. A higher prevalence of HBV infection was observed among males (33%) compared with females (12%; P < 0.05). Among women, a prior history of a sexually transmitted infection, injecting drug use (IDU), having had more than five lifetime sex partners, and having exchanged sex-for-goods were significantly associated with HCV infection, whereas an IDU history, syringe sharing, and having exchanged sex-for-goods were found to be associated with HBV infection. Among men, an IDU history and syringe/needle sharing were significantly associated with HCV infection. The IDU-related and sexual transmission of hepatitis viruses constitute a significant problem among young, HIV-infected, heterosexuals in Argentina.     

The role of the Purkinje network in premature ventricular complex-triggered ventricular fibrillation

The Purkinje network (PN) is the distal part of the ventricular conduction system, which has shown to play a central role in the pathophysiology of ventricular fibrillation (VF). Abnormal automaticity and triggered activity are commonly seen in the PN, and the resulting premature ventricular complexes (PVCs) are frequently recognized as triggers of this life-threatening arrhythmia. Catheter ablation targeting PN-related PVCs can be successfully performed in patients with medically refractory VF in a variety of arrhythmic substrates.