The natural history and risk factors for progression of non-alcoholic fatty liver disease and steatohepatitis

Nonalcoholic fatty liver disease (NAFLD) is a condition of increasing incidence in western Countries seldom associated to other diseases of high prevalence in general population (i.e. diabetes and obesity). NAFLD ranges from simple fatty liver to steatohepatitis (NASH), which may lead to cryptogenic cirrhosis and in some cases hepatocellular carcinoma (HCC). Natural history of NAFLD in humans is poorly understood and progression of liver disease seems to be due to interaction between hosting (i.e. genetic, gut flora, insulin resistance) and environmental factors (social and eating behaviours) that should be responsible of increased oxidative stress within hepatocytes. Even if we need non-invasive markers able to describe the progression of liver disease, only meaning of liver biopsy is useful to characterize the stigmata of worsening such as inflammation and fibrosis.     

Design and synthesis of the first generation of novel potent, selective, and in vivo active (benzothiazol-2-yl)acetonitrile inhibitors of the c-Jun N-terminal kinase

Several lines of evidence support the hypothesis that c-Jun N-terminal kinase (JNKs) plays a critical role in a wide range of diseases including cell death (apoptosis)-related disorders (neurodegenerative diseases, brain, heart, and renal ischemia, epilepsy) and inflammatory disorders (multiple sclerosis, rheumatoid arthritis, inflammatory bowel diseases). Screening of our internal compound collection for inhibitors of JNK3 led to the identification of (benzothiazol-2-yl)acetonitrile derivatives as potent and selective JNK1, -2, -3 inhibitors. Starting from initial hit 1 (AS007149), the chemistry and initial structure-activity relationship (SAR) of this novel and unique kinase inhibitor template were explored. Investigation of the SAR rapidly revealed that the benzothiazol-2-ylacetonitrile pyrimidine core was crucial to retain a good level of potency on rat JNK3. Therefore, compound 6 was further optimized by exploring a number of distal combinations in place of the chlorine atom. This led to the observation that the presence of an aromatic group, two carbons away from the aminopyrimidine moiety and bearing substituents conferring hydrogen bond acceptor (HBA) properties, could improve the potency. Further improvements to the biological and biopharmaceutical profile of the most promising compounds were performed, resulting in the discovery of compound 59 (AS601245). The in vitro and in vivo anti-inflammatory potential of this new JNK inhibitor was investigated and found to demonstrate efficacy per oral route in an experimental model of rheumatoid arthritis (RA).     

Reaction of 5-Aminosalicylic Acid with Peroxyl Radicals: Protection and Recovery by Ascorbic Acid and Amino Acids

Purpose The aims of the study are to analyze the interaction between 5-aminosalicylic acid (5-ASA) and peroxyl radicals and to evaluate the effect of some endogenous compounds such as ascorbic acid and amino acids on the oxidation of 5-ASA induced by 2,2′-azo-bis(2-amidinopropane) dihydrochloride. Methods The consumption and/or the recovery of 5-ASA (7.6 μM) exposed to a peroxyl radical source [2,2′-azo-bis(2-amidinopropane)] was followed by techniques such as spectrofluorescence, high-performance liquid chromatography, and differential pulse voltammetry. Results 5-Aminosalicylic acid was found to readily react with peroxyl radicals at micromolar concentrations and to protect c-Phycocyanin in a very similar fashion to that shown by Trolox. Exposure of 5-ASA to peroxyl radicals led to its oxidation into the corresponding quinone-imine. Disappearance of 5-ASA was prevented by tryptophan, cysteine, glutathione, and ascorbic acid. Furthermore, some of these compounds induced the partial (cysteine and glutathione) or total (ascorbic acid) recovery of 5-ASA when added after its almost total consumption. Conclusions 5-Aminosalicylic acid is a very efficient peroxyl radical scavenger. The 5-ASA oxidation by peroxyl radicals was prevented by ascorbic acid, cysteine, and glutathione. In addition, 5-ASA can be regenerated by these endogenous compounds, which would be a valuable mechanism to preserve 5-ASA in tissues undergoing oxidative stress conditions.     

Whole Body Protein Turnover and Net Protein Balance After Pediatric Thoracic Surgery: A Noninvasive Single‐Dose 15N Glycine Stable Isotope Protocol With End‐Product Enrichment

Background : We used the ¹⁵N glycine urinary end‐product enrichment technique to quantify whole body protein turnover following thoracic surgery. Materials and Methods : A single dose of ¹⁵N glycine (2 mg/kg) was administered orally on postoperative day 1 to children (1–18 years) following thoracic surgery. ¹⁵N enrichment of ammonia and urea was measured in mixed urine after 12 and 24 hours, respectively, and protein synthesis, breakdown, and net balance determined. Nitrogen balance (dietary intake minus urinary excretion) was calculated. Urinary 3‐methylhistidine:creatinine ratio was measured as a marker of skeletal muscle protein breakdown. Results : We enrolled 19 subjects—median (interquartile range): age, 13.8 years (12.2–15.1); weight, 49.2 kg (38.4–60.8)—who underwent thoracotomy (n = 12) or thoracoscopic (n = 7) surgery. Protein synthesis and breakdown by ¹⁵N enrichment were 7.1 (5.5–9) and 7.1 (5.6–9) g·kg^?1·d^?1 with ammonia (12 hours) as the end product, and 5.8 (3.8–6.7) and 6.7 (4.5–7.6) with urea (24 hours), respectively. Net protein balance by the ¹⁵N glycine and urinary urea nitrogen methods were ?0.34 (?0.47, ?0.3) and ?0.48 (?0.65, ?0.28) g·kg^?1·d^?1, respectively (r_s = 0.828, P < .001). Postoperative change in 3‐methylhistidine:creatinine ratio did not correlate significantly with protein breakdown or balance. Conclusion : The single‐dose oral administration of ¹⁵N glycine stable isotope with measurement of urinary end‐product enrichment is a feasible and noninvasive method to investigate whole body protein turnover in children. After major surgery, children manifest increased protein turnover and net negative balance due to increased protein breakdown.     

Evidence for the Presence of Non-Celiac Gluten Sensitivity in Patients with Functional Gastrointestinal Symptoms: Results from a Multicenter Randomized Double-Blind Placebo-Controlled Gluten Challenge

Non-celiac gluten sensitivity (NCGS) is characterized by the onset of symptoms after eating gluten-containing food. We aimed to single out NCGS subjects among subjects with functional gastrointestinal symptoms. Patients were enrolled in a multicenter double-blind placebo-controlled trial with crossover. Symptoms and quality of life were evaluated by means of 10-cm VAS and SF36. Iron parameters, transaminases and C reactive protein (CRP) were evaluated. After a three-week-long gluten-free diet (GFD), responsive patients were randomly assigned to gluten intake (5.6 g/day) or placebo for seven days, followed by crossover. The primary endpoint was the worsening of symptoms (VAS increase ≥3 cm) during gluten ingestion compared to placebo. One hundred and forty patients were enrolled and 134 (17 males, mean age 39.1 ± 11.7 years, BMI 22.4 ± 3.8) completed the first period. A total of 101 subjects (10 males, mean age 39.3 ± 11.0 years, BMI 22.3 ± 4.0) reported a symptomatic improvement (VAS score 2.3 ± 1.2 vs. 6.5 ± 2.2 before and after GFD, p = 0.001). 98 patients underwent the gluten challenge and 28 (all females, mean age 38.9 ± 12.7 years, BMI 22.0 ± 2.9) reported a symptomatic relapse and deterioration of quality of life. No parameters were found to be statistically associated with positivity to the challenge. However, 14 patients responded to the placebo ingestion. Taking into account this finding, about 14% of patients responding to gluten withdrawal showed a symptomatic relapse during the gluten challenge. This group is suspected to have NCGS.     

Bevacizumab in non small cell lung cancer: development, current status and issues

Bevacizumab is a monoclonal antibody directed against Vascular Endothelial Growth Factor (VEGF). Evidence about its efficacy in addition to first-line chemotherapy in non-small-cell-lung-cancer (NSCLC) has been produced by two large randomized phase III clinical trials (ECOG 4599 and AVAiL), conducted in a clinically selected population with non-squamous histology and without major risk factors for bleeding. In the ECOG 4599 trial, the addition of bevacizumab (15 mg/kg) to carboplatin plus paclitaxel produced a statistically significant and clinically relevant improvement in overall survival (OS), that was the primary endpoint of the trial (12.3 months vs 10.3 months, HR 0.79; p=0.003). Furthermore, patients receiving bevacizumab showed a significant improvement in progression-free survival (PFS) and in objective response rates. Treatment with bevacizumab was well tolerated by the majority of patients, but was still associated with increased risk of clinically significant bleeding (4.4% vs 0.7%, p0.001). In the AVAiL trial the addition of bevacizumab (at the dose of 7.5 and 15 mg/kg) to cisplatin plus gemcitabine produced a small improvement in PFS, but no differences in OS. Information from retrospective analysis and two large observational studies (SAIL and ARIES) have confirmed the safety profile of first-line bevacizumab with a wide range of chemotherapy partners, but whether its efficacy is comparable when combined with the different regimens is still unknown. The identification of predictive factors of efficacy would be relevant for the optimal use of the drug, but to date we have no conclusive data in this direction.     

Preserved Skeletal Muscle Mitochondrial Function,Redox State,Inflammation and Mass in Obese Mice with Chronic Heart Failure

Background: Skeletal muscle (SM) mitochondrial dysfunction, oxidative stress, inflammation and muscle mass loss may worsen prognosis in chronic heart failure (CHF). Diet-induced obesity may also cause SM mitochondrial dysfunction as well as oxidative stress and inflammation, but obesity per se may be paradoxically associated with high SM mass and mitochondrial adenosine triphosphate (ATP) production, as well as with enhanced survival in CHF. Methods: We investigated interactions between myocardial infarction(MI)-induced CHF and diet-induced obesity (12-wk 60% vs. standard 10% fat) in modulating gastrocnemius muscle (GM) mitochondrial ATP and tissue superoxide generation, oxidized glutathione (GSSG), cytokines and insulin signalling activation in 10-wk-old mice in the following groups: lean sham-operated, lean CHF (LCHF), obese CHF (ObCHF; all n = 8). The metabolic impact of obesity per se was investigated by pair-feeding ObCHF to standard diet with stabilized excess body weight until sacrifice at wk 8 post-MI. Results: Compared to sham, LCHF had low GM mass, paralleled by low mitochondrial ATP production and high mitochondrial reative oxygen species (ROS) production, pro-oxidative redox state, pro-inflammatory cytokine changes and low insulin signaling (p < 0.05). In contrast, excess body weight in pair-fed ObCHF was associated with high GM mass, preserved mitochondrial ATP and mitochondrial ROS production, unaltered redox state, tissue cytokines and insulin signaling (p = non significant vs. Sham, p < 0.05 vs. LCHF) despite higher superoxide generation from non-mitochondrial sources. Conclusions: CHF disrupts skeletal muscle mitochondrial function in lean rodents with low ATP and high mitochondrial ROS production, associated with tissue pro-inflammatory cytokine profile, low insulin signaling and muscle mass loss. Following CHF onset, obesity per se is associated with high skeletal muscle mass and preserved tissue ATP production, mitochondrial ROS production, redox state, cytokines and insulin signaling. These paradoxical and potentially favorable obesity-associated metabolic patterns could contribute to reported obesity-induced survival advantage in CHF.     

Oxygen Tension Modulates Inhibition of L-type Calcium Currents by Isoflurane in Human Atrial Cardiomyocytes

Background: Myocardial L-type Ca2+ currents (ICa,L) are inhibited by isoflurane in the presence of a partial pressure of oxygen (Po2) of 150 mmHg. In guinea pig cardiomyocytes, ICa,L are inhibited by reduced oxygen tensions. The authors therefore analyzed the effects of Po2 on ICa,L in human cardiomyocytes and the effects of isoflurane at reduced Po2.

Methods: Atrial cardiomyocytes were prepared from specimens of patients undergoing open-heart surgery and superfused with either a high or a low Po2 (150 or 12 +/- 1 mmHg) while ICa,L were measured with the whole cell patch clamp technique.

Results: Basal ICa,L were not changed by the Po2 (range, 9-150 mmHg) at 21[degrees] or 36[degrees]C. The reducing agent 1,4-dithiothreitol (DTT) left ICa,L unaffected, and the oxidizing agent 5,5'-dithio-bis(2-nitrobenzoic acid) (DTNB) irreversibly inhibited ICa,L. The Po2 significantly affected the inhibition of ICa,L by isoflurane (1 minimum alveolar concentration) that decreased ICa,L by 17 +/- 2.0% at the high Po2 but only by 5.8 +/- 2.9% (P = 0.037) at the low Po2. The inhibition of ICa,L by isoflurane was also significantly diminished (P = 0.018) by a low Po2 when isoflurane effects at both Po2 conditions were compared in the same cell.     

The service for Monitoring and Early Intervention against psychoLogical and mEntal suffering in young people (SMILE) at the University of L'Aquila: first year experience

Aim: To establish a comprehensive early intervention service for young people with mental suffering in L'Aquila, Italy, and to evaluate its effectiveness in delivering user friendly integrated interventions. Methods: The S ervice for M onitoring and early I ntervention against psycho L ogical and m E ntal suffering in young people (SMILE) began operation in November 2005 under the auspices of the Department of Mental Health, University of L'Aquila, Italy. It is the mission of our service to reduce the burden of mental suffering in young people by means of an earlier recognition of signs and symptoms, systematic evaluation of psychological distress and promotion of attitudes that encourage young people to seek care. We also aim to reduce the delays that young people at incipient risk of severe psychiatric illness experience in accessing appropriate psychiatric care. Results: Between November 2005 and November 2006, 216 young people (127 women, 89 men; mean age 21.8 ± 5.7 years) were referred for assessment. Thirty‐five per cent of patients had a diagnosis of severe mental illness (schizophrenia, bipolar disorder, incipient risk of psychosis). In addition, 80.5% of patients were admitted to the service more than once, totalling 685 admissions. Treatment modalities comprised cognitive behavioural treatment (61.5%), integrated psychosocial treatment (25.5%) and psychopharmacotherapy (15.9%). Conclusions: The SMILE service provides most of the mental health service requirements for young people with various forms of psychological suffering in Italy. Moreover, it offers appropriate case management with an early multimodal approach.