Modulatory effects of alpha-lipoic acid (ALA) administration on insulin sensitivity in obese PCOS patients.

PURPOSE: To evaluate the efficacy of alpha-lipoic acid (ALA) administration on hormonal and metabolic parameters of obese PCOS patients. METHODS: A group of 32 obese PCOS patients were selected after informed consent. 20 patients referred to have first grade relatives with diabetes type I or II. Hormonal and metabolic parameters as well as OGTT were evaluated before and after 12 weeks of ALA integrative administration (400 mg per os every day). RESULTS: ALA administration significantly decreased insulin, glucose, BMI and HOMA index. Hyperinsulinemia and insulin response to OGTT decreased both as maximal response (Δmax) and as AUC. PCOS with diabetes relatives showed the decrease also of triglyceride and GOT. Interestingly in all PCOS no changes occurred on all hormonal parameters involved in reproduction such as LH, FSH, and androstenedione. CONCLUSIONS: ALA integrative administration at a low dosage as 400 mg daily improved the metabolic impairment of all PCOS patients especially in those PCOS with familiar diabetes who have a higher grade of risk of NAFLD and predisposition to diabetes.

Asymptomatic bacteriuria in candidates for active treatment of renal stones: results from an international multicentric study on more than 2600 patients.

The occurrence of asymptomatic bacteriuria concomitant to urolithiasis is an issue for patients undergoing renal stone treatment. Disposing of a preoperative urine culture is essential to reduce the risk of septic events. The endpoint of the study is to report which characteristics of candidates for renal stone treatment are frequently associated with positive urine culture. 2605 patients were retrospectively enrolled from 14 centers; inclusion criteria were age > 18 and presence of a single renal stone 1-2 cm in size. The variables collected included age, gender, previous renal surgery, comorbidities, skin-to-stone distance, stone size, location, density, presence of hydronephrosis. After a descriptive analysis, the association between continuous and categorical variables and the presence of positive urine culture was assessed using a logistic regression model. Overall, 240/2605 patients (9%) had preoperative bacteriuria. Positive urine culture was more frequent in females, patients with previous renal interventions, chronic kidney disease, congenital anomalies, larger stones, increased density. Multivariate analysis demonstrated that previous renal interventions (OR 2.6; 95% CI 1.9-3.4; p < 0.001), renal-related comorbidities (OR 1.31; 95% CI 1.19-1.4; p < 0.001), higher stone size (OR 1.06; 95% CI 1.02-1.1; p = 0.01) and density (OR 1.00; 95% CI 1.0-1.00; p = 0.02) were associated with bacteriuria; male gender and lower caliceal location were inversely related to it. Beyond expected risk factors, such as female gender, other parameters are seemingly favoring the presence of positive urine culture. The awareness of variables associated with bacteriuria allows to assess which individuals are at increased risk of presenting bacteriuria and reduce the rate of septic complications.

Reduced susceptibility to quinupristin/dalfopristin in Enterococcus faecium in Greece without prior exposure to the agent.

During 2005-2006, a total of 865 Enterococcus faecium isolated from patients from eight Greek hospitals were tested for susceptibility to quinupristin/dalfopristin (Q/D). Among them, 250 genetically unrelated strains (28.9%) were found to be intermediate-resistant to Q/D (minimum inhibitory concentration (MIC) 1.5-4 mg/L); all were resistant to dalfopristin (MIC=16-64 mg/L), whilst 69% were resistant to quinupristin, carrying the ermB gene. No strain was found to carry any of the known genes, such as vatE and vatD, involved in Q/D resistance, indicating that a non-transferable undetermined mechanism is responsible for the expression of low-level Q/D resistance. The high percentage of Q/D-intermediate-resistant E. faecium in Greece was not associated with prior consumption of the agent or with the veterinary use of virginiamycin.

Aqueous extract of Arbutus unedo inhibits STAT1 activation in human breast cancer cell line MDA-MB-231 and human fibroblasts through SHP2 activation.

Arbutus unedo L. has been for a long time employed in traditional and popular medicine as an astringent, diuretic, urinary anti-septic, and more recently, in the therapy of hypertension and diabetes. Signal transducer and activator of transcription 1 (STAT1) is a fascinating and complex protein with multiple yet contrasting transcriptional functions. Although activation of this nuclear factor is finely regulated in order to control the entire inflammatory process, its hyper-activation or time-spatially erroneous activation may lead to exacerbation of inflammation. The modulation of this nuclear factor, therefore, has recently been considered as a new strategy in the treatment of inflammatory diseases. In this study, we present data showing that the aqueous extract of Arbutus unedo's leaves exerts inhibitory action on interferon-gamma (IFN-gamma) elicited activation of STAT1, both in human breast cancer cell line MDA-MB-231 and in human fibroblasts. This down-regulation of STAT1 is shown to result from a reduced tyrosine phosphorylation of STAT1 protein. Evidence is also presented indicating that the inhibitory effect of this extract may be mediated through enhancement of tyrosine phosphorylation of SHP2 tyrosine phosphatase. The modulation of this nuclear factor turns out into the regulation of the expression of a number of genes involved in the inflammatory response such as inducible nitric oxide synthase (iNOS) and intercellular adhesion molecule-1 (ICAM-1). Taken together, our results suggest that the employment of the Arbutus unedo aqueous extract is promising, at least, as an auxiliary anti-inflammatory treatment of diseases in which STAT1 plays a critical role.

The role of constitutive and inducible nitric oxide synthase in the human brain after subarachnoid hemorrhage.

AIM: Results of prior experimental studies show that nitric oxide (NO) plays an important role in the pathogenesis of vasospasm. In the present study, the expression of endothelial NO synthase (eNOS), neuronal NO synthase (nNOS) and of inducible NO synthase (iNOS) in the human brain after subarachnoid haemorrhage were studied. METHODS: Twenty-three samples of gyrus rectus or temporal operculum that were obtained during a surgical approach to anterior circulation aneurysms were used for this study. Seven samples were obtained during surgery from patients who underwent operation for unruptured aneurysms (control group). eNOS-mRNA, nNOS-mRNA and iNOS-mRNA were extracted and amplified by RT-PCR. Patients were subdivided for intergroup comparison by: age < 60 / > 60 years; source of sample; clinical grading; extent of subarachnoid clot; presence of intracerebral/intraventricular hematoma; surgical timing; vasospasm; outcome. RESULTS: There was a significant increase in the expression of eNOS between SAH and control groups (P=0.046); eNOS hyperexpression was higher in the patients in poor clinical conditions (P=0.002) and lasted until the late phase of haemorrhage. nNOS overall expression was unchanged but hyperexpression was observed in the patients in poor clinical conditions (P=0.008). There was a significant hyperexpression of iNOS in SAH group (P=0.026), and in patients with vasospasm (P=0.0024); the expression was significantly reduced in the late phase of haemorrhage (P=0.0038). CONCLUSIONS: The acute decrease of NO after SAH is not determined by reduced constitutive NOS expression and iNOS induction is a consequence of SAH and plays a major role in the pathogenesis of vasospasm.

Activation of human monocyte-derived macrophages by interferon gamma is accompanied by increase of poly(ADP-ribose) polymerase activity.

We have investigated poly(ADP-ribosyl)ation processes in human monocyte-derived macrophages and the effect of the activating cytokine, interferon gamma (IFN-gamma) on these processes. IFN-gamma was shown to increase the activity of poly(ADP-ribose) polymerase in human macrophages. A 2-3-fold enhancement of poly(ADP-ribose) polymerase activity was observed after 3-4 h of incubation with IFN-gamma, whose effects were dose-dependent and maximal at 20-50 U/ml. Staining with anti-poly(ADP-ribose) antibodies and purification of ADP-ribosylated nuclear proteins by affinity chromatography over boronate agarose showed that enhancement of poly(ADP-ribose) polymerase activity by IFN-gamma was accompanied by accumulation of poly(ADP-ribose) polymers in nuclear proteins. The effects of IFN-gamma on poly(ADP-ribose) polymerase activity were not due to an enhanced accumulation of the message for the enzyme, indicating that the activation of the enzyme activity was due to post-transcriptional modifications. IFN-gamma was shown to induce DNA strand breaks in human macrophages. This phenomenon followed the same time-course and was evident with the same doses of IFN-gamma that increased poly(ADP-ribose) polymerase activity. Since poly(ADP-ribose) polymerase is known to require DNA nicks for its activity, the capability of IFN-gamma to induce DNA strand breaks can explain its effects on poly(ADP-ribosyl)ation processes.

Changes in activity and mRNA levels of poly(ADP-ribose) polymerase during rat liver regeneration.

ADP-ribosylation of nuclear proteins, catalysed by the enzyme poly(ADP-ribose) polymerase, is involved in the regulation of different cellular processes of DNA metabolism. To further clarify the role of the enzyme during proliferating activity of mammalian cells, we have studied the control of gene expression in regenerating rat liver. The changes in activity and mRNA levels were analysed during the early and late phases of the compensatory model. When enzyme activity was measured in isolated liver nuclei obtained at different times after hepatectomy, two different phases were observed: an early wave occurring before the onset of DNA synthesis, and a second one, starting several hours after the onset of DNA synthesis and returning to control values at later times. The evaluation of the enzymatic level in nuclear extracts and by activity gel analysis showed a more gradual increase starting 1 day after hepatectomy, in concomitance with the peak of DNA synthesis. By using a specific murine cDNA probe, a significant enhancement of mRNA levels for poly(ADP-ribose) polymerase was observed during liver regeneration, slightly preceding the onset of DNA synthesis. The results obtained show that changes in poly(ADP-ribose) polymerase activity, during liver regeneration, are associated both to early events preceding the increase in DNA synthesis and to later phases of the cell proliferation process.

Increase of poly(ADP-ribose) polymerase mRNA levels during TPA-induced differentiation of human lymphocytes.

The non-mitogenic stimulation of human peripheral blood mononuclear cells (PBMC) with low concentrations of the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA) caused a progressive increase in the percent fraction of the cells that were positive for the early activating antigen CD69. At the same time, it caused a progressive increase in the steady-state levels of poly(ADP-ribose) polymerase (pADPRP) transcripts. A further increase in TPA concentration, while inducing the maximal expression of the levels of CD69 activating surface antigen, both in the presence or in the absence of proliferative activity, did not evoke any additional hightening of pADPRP mRNA levels. Time course of PBMC stimulation with a non-mitogenic dose of TPA showed an early increase in the accumulation of pADPRP mRNA, which changed at 8-16 h, and remained high for several days thereafter. On the basis of these data, we suggest that the increase in pADPRP mRNA may be associated with the commitment of human lymphocytes from a quiescent (G0) to an activated (G1) state.

TPA and cycloheximide modulate the activation of NF-kappa B and the induction and stability of nitric oxide synthase transcript in primary neonatal rat hepatocytes.

12-O-Tetradecanoylphorbol 13-acetate (TPA) elicited a transient increase in the transcription of the inducible nitric oxide synthase (iNOS) gene coupled with a shortening of the half-life of its mRNA in primary neonatal rat hepatocytes. These effects of TPA were preceded by a surge in nuclear translocation of the transcription factor NF-kappa B, and followed by a mounting accumulation of NO-2 in the growth medium. Even cycloheximide (CHX) added by itself elicited an early, sustained activation of NF-kappa B followed by an intense induction of iNOS gene expression, irrespective of what degree of protein synthesis inhibition was brought about by the several concentrations tested. When given together, TPA and CHX exerted additive effects on hepatocellular iNOS mRNA levels. These results suggest the likelihood of an ordered sequence of events by which an activated NF-kappa B mediates the induction of iNOS gene expression in TPA- and/or CHX-treated primary hepatocytes.

Involvement of DNA polymerase beta in proliferation of rat liver induced by lead nitrate or partial hepatectomy.

We have studied the expression pattern of DNA polymerase beta in two different models of in vivo cell proliferation. Both mRNA levels and enzyme activity of DNA polymerase beta markedly increased before and/or during DNA synthesis in proliferating hepatocytes in mitogen-treated and partially hepatectomized rats. The time-courses of the expression of the gene coding for DNA polymerase beta were significantly different in the two cell systems. A 5-fold increase in DNA polymerase beta mRNA was observed 8 h after lead nitrate administration, i.e. well before the onset of DNA synthesis. In the regenerative liver cells a 3-fold increase in the amount of mRNA was observed 24-48 h after partial hepatectomy, the event being coincident with extensive DNA synthesis. In both systems, the increase of mRNA levels was always paralleled by an increase in enzyme activity, suggesting that DNA polymerase beta activity may be regulated at a pre-translational level.

Protective effect of NO on gastric lesions and inhibition of expression of gastric inducible NOS by flurbiprofen and its nitro-derivative, nitroflurbiprofen.

Nitroflurbiprofen (NFP) causes significantly less gastric lesions than flurbiprofen (FP), probably because of its capacity to release nitric oxide (NO) in the stomach. Lipopolysaccharide (LPS), which induces the expression of an inducible type of NO synthase (iNOS) in rat stomach, also reduces gastric mucosal damage elicited by FP. Furthermore, both FP and NFP decrease significantly the amount of mRNA encoding iNOS induced by LPS in the stomach. The inhibitory effect of NFP seems to be due at least in part to its ability to release NO.

Differential expression pattern of jun B and c-jun in the rat brain during the 24-h cycle.

Data from a previous report [3] demonstrated that the proto-oncogene c-fos mRNA expression undergoes basally a circadian fluctuation in the rat brain. The present study was designed to verify by means of Northern blot hybridization the eventual occurrence of a spontaneous oscillation in the expression of other two proto-oncogenes, jun B and c-jun, during 24 h. Rats were either entrained to a light-dark photoperiod or maintained under constant darkness or light. During the dark period, as well as the subjective night, the jun B mRNA levels in the cerebral cortex and striatum were 4-6 times higher than in the light hours or subjective day. No consistent oscillation was found in the c-jun mRNA expression during 24 h in any of the examined brain regions. These results suggest the possibility of different interactions of the c-fos, jun B and c-jun gene products throughout a 24-h period in discrete brain regions.

Biotransformation and cytotoxic properties of NO-donors on MCF7 and U251 cell lines.

Previous studies have shown a role for nitric oxide (NO) as a cytotoxic effector. In the present work, two chemically different NO-donors such as glyceryl trinitrate (GTN) and S-nitroso-N-acetylpenicillamine (SNAP) were evaluated for both NO release and cytostatic/cytotoxic properties. Nitrite accumulation in the supernatant of MCF-7 and U251 cell lines indicated a greater and quickly release of NO derived from SNAP. A time-course of hemoglobin absorption spectral changes showed a greater release of NO derived from GTN in presence of cells compared to the values observed in the media, confirming that the release of NO by GTN can be enzymatic and non-enzymatic. On the contrary, SNAP generated NO without contribution of cellular components and saturated oxyhemoglobin quickly, within 2 hours. Both NO-donors inhibited thymidine incorporation in a similar manner and dose-dependently in U251 cells, but not in MCF-7 cells, where SNAP at the highest tested dose of 1000 microM induced only a 33% cytostatic effect. About trypan blue exclusion test, after 24 h GTN and SNAP, releasing similar amounts of NO, showed comparable cytotoxic effects on U251 cells (50% dead cells), but not on MCF-7 cells, where GTN resulted more cytotoxic. From our data, the "in vitro" antitumoral activity of NO-donors seems to be related to the type of tumor cell lines, to the amount and duration of NO release.

[The putative role and use of DHEA and its association with the hormone replacement therapy]. / Dall'uso del solo DHEA alla sua associazione con la terapia ormonale sostitutiva.

The putative role and use of dehydroepiandrosterone (DHEA) as replacement therapy for menopausal women has been under consideration during the latest years. DHEA is one of the main adrenal hormones that progressively reduces its plasmatic levels from the beginning of ageing. This phenomenon implies not only the reduction of the plasmatic androgens but also the decrease of a peculiar category of hormones, named neurosteroids, in particular one: allopregnanolone. This review aims to elucidate the peculiar aspects of DHEA administration and its putative use as substitutive/integrative hormonal treatment alone or in combination with the traditional hormone replacement therapy.

Release of bone markers in immediately loaded and nonloaded dental implants: a randomized clinical trial.

The aim of this study was to compare the release of bone markers during osseointegration of immediately loaded and nonloaded implants. Forty patients who were indicated for rehabilitation with dental implants randomly received either implant and prosthesis placement within 72 hours (group IM) or implant insertion and no prosthesis placement (group NL). Peri-implant crevicular fluid was collected immediately after implant insertion and 7, 15, 30, 60, 90, and 120 days after surgery and levels of osteoprotegerin, transforming growth factors, osteocalcin, osteopontin, and parathyroid hormone were evaluated using Luminex assay. Bleeding index and peri-implantar sulcus depth were also evaluated. The data were compared using statistical tests (α = 5%). No statistical difference was found regarding demographic and clinical parameters (p > .05). Transforming growth factors, osteoprotegerin, osteopontin, and parathyroid hormone presented an earlier release peak in group IM than in NL group (p < .05). Osteocalcin achieved higher levels in group IM versus group NL between 7 and 30 days of evaluation (p < .05). It may be concluded that earlier loading positively modulates bone mediators release around immediately loaded implants when compared with nonloaded dental implants.

Use of mechanical devices for the intermaxillary registration in edentulous patients treated with implants.

This article describes the use of three devices that replace the wax rims in the registration of the intermaxillary position and of the vertical dimension of occlusion in completely edentulous patients who have been treated with Brånemark implants. The devices consist of a mechanical "tooth" that can be adjusted in all three dimensions of space and of two plates that support the registration material. The mechanical tooth is connected to an abutment in the anterior region and a contact is established with a tooth in the opposite jaw at the vertical dimension in which the patient will be restored. This allows the operator to position the mandible in the centric relation in a condition of neuromuscular deprogramming and in the absence of posterior interferences. The two metal plates are then fixed to the posterior abutments, one on each side, and support the wax and zinc oxide eugenol paste used to register the intermaxillary position just established.

[Influence of transcutaneous stimulation (TENS) of the posterior tibial nerve on various cysto-manometric parameters in patients with neurologic bladder. Initial data on 6 patients]. / Influenze della stimolazione transcutanea (TENS) del tibiale posteriore su alcuni parametri cistomanometrici in pazienti con vescica neurologica. Primi dati su sei pazienti.

The authors report about their first study of the detrusor activity modifications induced by electrical stimulation of the posterior tibial nerve in six patients with neurogenic bladder. The results suggest that this new application inhibit detrusor activity.

Mycoplasma synoviae in the guinea-fowl.

A mycoplasma strain, isolated from natural synovitis of guinea-fowl, was identified as Mycoplasma synoviae. In experimental infection this agent showed clear pathogenicity for guinea-fowl, and to a lesser degree for chickens. In guinea-fowl the strain was more likely to result in synovitis and amyloidosis when inoculated by the intravenous route and to produce sinusitis after intrasinusal inoculation.

Effects of kainic acid lesioning on poly(ADP-ribose) polymerase (PARP) activity in the rat striatum in vivo.

Poly(ADP-ribose) polymerase (PARP) is activated in glutamate-induced toxicity of neurons in culture (Cosi et al., 1994). Since injection of the excitatory amino acid, kainic acid (KA) into the rat striatum induces a delayed neuronal death, the effects of this in vivo excitotoxin lesioning procedure on striatal PARP activity was investigated. PARP activity was measured in striatal extracts both in the absence ("endogenous" activity) and presence ("total" activity) of exogenously-added fragmented DNA. KA (5nmols/1microl) produced significant and time-dependent changes in striatal PARP activity, compared to saline-injected control animals: no changes at 6h after intrastriatal KA, a 68% and 48% decrease in endogenous and total PARP activity respectively at 12h, a doubling in endogenous PARP activity at 24h, and a 382% and 60% increase in endogenous and total activities at 1 week after KA. PARP cleavage was not detected at any time point. These results suggest a participation of PARP in KA-induced toxicity in the brain in vivo.