Identification and analysis of murine pancreatic islet enhancers.

AIMS/HYPOTHESIS: The paucity of information on the epigenetic barriers that are blocking reprogramming protocols, and on what makes a beta cell unique, has hampered efforts to develop novel beta cell sources. Here, we aimed to identify enhancers in pancreatic islets, to understand their developmental ontologies, and to identify enhancers unique to islets to increase our understanding of islet-specific gene expression. METHODS: We combined H3K4me1-based nucleosome predictions with pancreatic and duodenal homeobox 1 (PDX1), neurogenic differentiation 1 (NEUROD1), v-Maf musculoaponeurotic fibrosarcoma oncogene family, protein A (MAFA) and forkhead box A2 (FOXA2) occupancy data to identify enhancers in mouse islets. RESULTS: We identified 22,223 putative enhancer loci in in vivo mouse islets. Our validation experiments suggest that nearly half of these loci are active in regulating islet gene expression, with the remaining regions probably poised for activity. We showed that these loci have at least nine developmental ontologies, and that islet enhancers predominately acquire H3K4me1 during differentiation. We next discriminated 1,799 enhancers unique to islets and showed that these islet-specific enhancers have reduced association with annotated genes, and identified a subset that are instead associated with novel islet-specific long non-coding RNAs (lncRNAs). CONCLUSIONS/INTERPRETATIONS: Our results indicate that genes with islet-specific expression and function tend to have enhancers devoid of histone methylation marks or, less often, that are bivalent or repressed, in embryonic stem cells and liver. Further, we identify a subset of enhancers unique to islets that are associated with novel islet-specific genes and lncRNAs. We anticipate that these data will facilitate the development of novel sources of functional beta cell mass.

Similar serum levels of IL-6 and its soluble receptors in patients with HCV-related arthritis and rheumatoid arthritis: a pilot study.

The high serum levels of Interleukin-6 (IL-6) and its soluble receptors (sIL-6r and sgp130), described in the course of Rheumatoid Arthritis (RA), have been linked to the enhanced activity of this cytokine in this disorder. In this study, the serum concentrations of IL-6 and its soluble receptors were determined in a group of patients with HCV-related arthritis (HCVrA), a condition resembling RA in several aspects, and then compared to those found in a sample of subjects affected by RA. Twenty-one patients with HCVrA, 24 patients with RA and 20 healthy subjects (control group) were examined. Different ELISA methods were used for determination of serum concentrations of IL-6, sIL-6r and sgp130. Increased IL-6 serum levels were found in 15 (71 %) of the patients with HCVrA and in 16 (62 %) of those with RA. Eight (38 %) of the patients with HCVrA and 11 (46%) of those with RA denoted high levels of sIL-6r, while sgp130 levels were elevated in 21 (76%) of the patients with HCVrA and in 16 (69%) of those with RA. A significant difference between the median values of sIL-6r and sgp130 levels in the two groups of patients versus controls was found. A mild correlation of these parameters with RF levels was detected in the RA group. Furthermore, in HCVrA patients the serum levels of IL-6, sIL-6r and sgp130 appeared unrelated to HCV viraemia and to levels of transaminases. The enhanced serum levels of IL-6 in HCVra patients indicate an increased synthesis and hyperactivity of this cytokine in HCVrA, and the substantial similarity of the behaviour of IL-6 and its serum receptors in the two groups of patients suggests common mechanisms with RA, in which the function of I L-6 is central.

Circulating levels of interferon-gamma in course of hepatitis C virus-related arthritis.

The aim was to weigh the serum concentrations of interferon gamma (IFN-gamma), a cytokine that enhances Th1-cell differentiation and suppresses collagen synthesis and angiogenesis, in two apparently distinct diseases, hepatitis C virus-related arthritis (HCVrA) and rheumatoid arthritis (RA), which share some overlapping immunological features. In this study, IFN-gamma serum levels were assayed by an ELISA method in 21 HCVrA patients and in 16 with RA. Very low IFN-gamma serum levels were found in five out of 21 patients with HCVrA and only in three out of 16 RA patients. Median value (range) resulted decrease in both HCVrA and RA groups, that is, 0.29 (0.04-1.49) versus 0.20 (0.05-1.18) IU/mL, P = 0.58. No correlation was evidenced with hepatic and arthritic involvements, nor between IFN-gamma serum levels and viral replication and moreover with the positivity of antinuclear antibody, rheumatoid factor, and anti-cyclic citrullinated peptides antibodies. These results show that IFN-gamma behavior appears similar in HCVrA and RA groups reinforcing the lack of significant differences between HCVrA and RA patients. Low circulating levels could be explained with the fact that IFN-gamma is not an isolate cytokine, but a piece of composite system regulated in a complex fashion, with many different factors contributing.

A new strategy for the early diagnosis of rheumatoid arthritis: a combined approach.

Rheumatoid arthritis [RA] is one of the most common and severe autoimmune rheumatic diseases, diagnosed primarily according to clinical manifestations and radiological reports. For many years, laboratory diagnosis of rheumatoid arthritis has relied on the detection of rheumatoid factor [RF], as established by the ACR criteria. A recent test to detect antibodies towards citrullinated peptides, called the anti-CCP assay, showed a similar sensitivity but a more elevated specificity than the RF test. Our intention was the recognition of an optimal diagnostic strategy that exhibits the highest sensitivity and specificity for RA detection. To this purpose, we examine the usefulness of autoantibodies in RA testing, evaluating the diagnostic performance of conventional and innovative assays for RF detection, and ELISA anti-CCP test, for anti-CCP antibodies detection, by a prospective study. Multiplex cytofluorimetric test appeared to be more sensitive and specific than nephelometric assay for RF detection. Hence, a novel combined approach, significantly increasing the diagnostic sensitivity for RA, was planned, employing the multiplex RF test in combination with the anti-CCP test.

Usefulness of a tilt training program for the prevention of refractory neurocardiogenic syncope in adolescents: A controlled study.

BACKGROUND: Recurrent syncope represents a debilitating disorder and quality of life deteriorates as a function of recurrence of symptoms. Although the administration of beta-blockers, vasoconstrictors, fludrocortisone, and serotonin reuptake inhibitors may be helpful in preventing episodes, many patients are intolerant of or respond poorly to these agents. Orthostatic training has been reported to be effective in preventing refractory syncope. Thus, to determine whether a tilt training program could prevent symptoms in adolescents, the following controlled study was undertaken. METHODS AND RESULTS: Forty-seven consecutive adolescents (18 male and 29 female, mean age 16.0+/-2.2 years) with recurrent syncope and positive head-up tilt test refractory to previous traditional therapies were distributed between 2 groups, depending on their consent (24 patients) or refusal (controls, 23 patients) to enter the program. Orthostatic training was started, in the presence of a family member, with a series of 5 in-hospital sessions. The 24 patients and their relatives were then instructed to perform the tilt training at home by standing against a wall twice a day for a planned duration of up to 40 minutes, depending on the in-hospital orthostatic tolerance. Head-up tilt response was reevaluated after 1 month, and the clinical effect was noted over a mean follow-up of 18. 2+/-5.3 months (range 15 to 23); 26.1% of patients in the control group and 95.8% of patients in the training group became tilt-negative (P<0.0001). Spontaneous syncope was observed in 56.5% versus 0% in the control and training group, respectively (P<0.0001). CONCLUSIONS: Orthostatic training was found to significantly improve symptoms of adolescents with neurocardiogenic syncope unresponsive to or intolerant of traditional medications. Twice-a-day training sessions of 40 minutes were well accepted by patients.

Effects of paroxetine hydrochloride, a selective serotonin reuptake inhibitor, on refractory vasovagal syncope: a randomized, double-blind, placebo-controlled study.

OBJECTIVES: The purpose of the study was to determine whether the well tolerated serotonin reuptake inhibitor paroxetine hydrochloride could prevent vasovagal syncope in patients resistant to or intolerant of previous traditional therapies. BACKGROUND: Serotonergic mechanisms play a major role in the processes leading to neurocardiogenic vasovagal syncope, and serotonin reuptake inhibitors have been reported to be effective in preventing refractory syncope. METHODS: Sixty-eight consecutive patients (26 men and 42 women, mean age 44.7+/-16.5 years) with recurrent syncope and positive head-up tilt test and in whom standard therapies with beta-adrenergic blocking agents, vagolytic, negative inotropic or mineral corticoid agents were ineffectual or poorly tolerated were referred for study. Patients randomly received either paroxetine at 20 mg once a day or a placebo. A head-up tilt test was then reperformed after one month of treatment, and the clinical effect was noted over a mean follow-up of 25.4+/-7.9 months. RESULTS: The response rates (negative tilt test) after one month of treatment were 61.8% versus 38.2% (p < 0.001) in the paroxetine and placebo groups, respectively. During follow-up spontaneous syncope was reported in six patients (17.6%) in the paroxetine group as compared to 18 patients (52.9%) in the placebo group (p < 0.0001). Only one patient (2.9%) asked to be discontinued from the drug for severe side effects. CONCLUSIONS: Paroxetine was found to significantly improve the symptoms of patients with vasovagal syncope unresponsive to or intolerant of traditional medications and was well tolerated by patients.

Sedimentable mineral organic detritus as radioecological indicator.

The sedimentable mineral organic detritus (SMOD) drawn by rivers can be considered an important matrix for the monitoring of contaminants in aquatic environments. In Italy the collection and radioactivity analysis of SMOD has been introduced and standardised in the 80s. However hydrological parameters, like the flow and the amount of suspended matter close to the sampling points, must be considered to improve the methodology. This technique has been applied by the authors since 1992 to monitor the concentration of both 137Cs (following the Chernobyl accident) and 131I (because of possible waste discharge from Perugia University Hospital) along the Umbrian course of the Tiber River. In this paper the results of 137Cs water concentration are presented and discussed in the light of our interpretative working hypothesis based on hydrological parameters. This work allowed us to reduce the number of sampling points without loss of radioecological information.

[Effectiveness of paroxetine in the treatment of refractory vasovagal syncope in young patients]. / Efficacia della paroxetina nel trattamento della sincope vasovagale refrattaria in pazienti giovani.

OBJECTIVES: The aim of the study was to assess whether the well-tolerated serotonin re-uptake inhibitor paroxetine hydrochloride could prevent vasovagal syncope in young patients resistant to or intolerant of previous traditional therapies. BACKGROUND: Serotonergic mechanisms may play a major role in the pathophysiology of neurocardiogenic syncope, and serotonin re-uptake inhibitors have been recently reported to be effective in preventing episodes. METHODS: Forty-one consecutive young patients (13 male and 28 female), aged less than thirty years with recurrent syncope and positive head-up tilt test, and in whom standard therapies with beta-blocking, vagolytic, negative inotropic or mineral corticoid agents were ineffectual, poorly tolerated or contraindicated, randomly received either paroxetine at 20 mg once a day or a placebo. A head-up tilt test was then re-performed after one month of treatment, and the clinical effect was noted over a mean follow-up of 27.1 +/- 6.6 months. RESULTS: The response rates (negative tilt test) after one month of treatment were 57.1 versus 33.3% (p < 0.001) in the paroxetine and placebo groups, respectively. During follow-up, spontaneous syncope was observed in 4 patients (19%) in the paroxetine group and in 12 patients (60%) in the placebo group (p < 0.001). Only one patient (4.8%) asked to be discontinued from the drug for severe recurrent headache. CONCLUSIONS: Paroxetine significantly improved symptoms of young patients with recurrent vasovagal syncope unresponsive to or intolerant of traditional medications and was well tolerated by patients.

[Evaluation of the effects of diverse therapeutic treatments versus no treatment of patients with neurocardiogenic syncope]. / Valutazione degli effetti di diversi trattamenti terapeutici rispetto a nessun trattamento in pazienti con sincope neurocardiogena.

Head-up tilt test was introduced in clinical practice to assess vasovagal syncope and its use has further been extended to evaluate the efficacy of drug administration in these patients. Nevertheless, the effects of tilt test on vasovagal syncope have never been compared with those obtained by ethylephrine or propranolol administration. One hundred and sixty-nine consecutive patients with vasovagal syncope and positive baseline or nitrate-potentiated tilt test (60 degrees upright position for 45 min, or until syncope occurred; 5 mg sublingual isosorbide dinitrate administration if no symptoms occurred) were randomly distributed among three groups: Group A (57 control patients discharged without medical therapy); Group B (56 patients discharged with 75 mg/die ethylephrine); Group C (56 patients discharged with 80 mg/die propranolol). Tilt test was repeated after 1 month, while clinical outcome was evaluated monthly for a mean follow-up of 37.1 +/- 15.6 months. No significant differences in acute tilt-induced syncope recurrence rates were obtained among groups at test repetition since 70.2% of Group A, 69.6% of Group B and 62.5% of Group C experienced syncope. At 3-year follow-up 82.4% of Group A, 83.9% of Group B and 87.5% of Group C (NS among groups) remained symptom free, the most important clinical result being obtained in untreated patients. These data suggest that tilt test execution may prevent syncope recurrence as ethylephrine or propranolol administration. Irrespective of the therapeutical choice, the "controlled reproduction" of symptoms and some psychophysical training of patients to avoid precipitating circumstances, to recognize early symptoms promptly to be reverted by Trendelemburg position, may produce the same clinical improvement as (empiric) ethylephrine or propranolol therapy.

Cytotoxic activity and IL-1 production in mice infected with Aspergillus niger.

In this report, we demonstrate an interaction between macrophages and T lymphocytes during A. nigr infection. Supernatants obtained after 48 hrs adherence of infected peritoneal macrophages were able to increase the cytotoxicity of T lymphocytes. Our results also indicate that macrophage supernatant (MS) from mice, in the first 5 days after challenge, is more active on T cell than MS produced later. Splenic T cells activated by IL-1 from mice at 5 days of infection show a significantly increased cytotoxicity, at 10 days after challenge, the cytotoxicity of T cells activated by IL-1 did not significantly differ from non-activated T cells.

Uptake of radiocesium by mushrooms.

After the Chernobyl accident in April 1986 the 134Cs, 137Cs and 40K activity of mushrooms of different genera and their corresponding soils from woods located in the province of Umbria (Italy) was determined. The results indicated: a temporally increasing trend for 137Cs up to a "limit value" depending on local fallout in 1986; different cesium concentrations in various mushroom genera; and uptake of potassium from soil that did not appear to compete with cesium uptake. No reliable correlation can be inferred between the cesium content of the soil and those of mushrooms.

Interleukin-2 and increased natural killer activity in mice experimentally infected with Aspergillus niger.

The authors studied the production and effect of interleukin 2 obtained from T lymphocytes stimulated with Aspergillus niger on populations of LGL cells from the peripheral blood and spleen of infected and non-infected mice. The results show an increase in IL-2 production by T lymphocytes in the spleen. In addition, the data demonstrate an increase in LGL activity in the blood and spleen of infected mice. This was seen as the percentage of growth inhibition towards Aspergillus niger, around the tenth day of infection, and reached a maximum of activity on the fifteenth day. The NK activity of the LGL studied increases both in infected and non-infected mice when IL-2 from T lymphocytes of infected mouse blood and spleen is present.

Modification of the T, K and NK cell population in experimental infection of mice with Aspergillus niger.

A study has been made of the cellular response by T, K and NK cells in mice experimentally infected with Aspergillus niger. The tests were made on mice treated with 5 X 10(5) conidia. The lymphocyte populations in the blood, spleen and pulmonary lymphnodes have studied to identify possible modifications in immune response in time. The results obtained show that in experimentally infected mice with respect to uninfected mice there is an increase in T-lymphocytes, particularly in the spleen, a percentage increase in EA-RFC cells in the blood, spleen and lymphnodes, a percentage increase in LGL cells, with a rise in associated NK activity, in the blood and spleen in the first days of infection, but a delayed increase in the lymphnodes.