Enteric α-synuclein impairs intestinal epithelial barrier through caspase-1-inflammasome signaling in Parkinson's disease before brain pathology.

Bowel inflammation, impaired intestinal epithelial barrier (IEB), and gut dysbiosis could represent early events in Parkinson's disease (PD). This study examined, in a descriptive manner, the correlation among enteric α-synuclein, bowel inflammation, impairments of IEB and alterations of enteric bacteria in a transgenic (Tg) model of PD before brain pathology. Human A53T α-synuclein Tg mice were sacrificed at 3, 6, and 9 months of age to evaluate concomitance of enteric inflammation, IEB impairments, and enteric bacterial metabolite alterations during the early phases of α-synucleinopathy. The molecular mechanisms underlying the interplay between α-synuclein, activation of immune/inflammatory responses and IEB alterations were investigated with in vitro experiments in cell cultures. Tg mice displayed an increase in colonic levels of IL-1ß, TNF, caspase-1 activity and enteric glia activation since 3 months of age. Colonic TLR-2 and zonulin-1 expression were altered in Tg mice as compared with controls. Lipopolysaccharide levels were increased in Tg animals at 3 months, while fecal butyrate and propionate levels were decreased. Co-treatment with lipopolysaccharide and α-synuclein promoted IL-1ß release in the supernatant of THP-1 cells. When applied to Caco-2 cells, the THP-1-derived supernatant decreased zonulin-1 and occludin expression. Such an effect was abrogated when THP-1 cells were incubated with YVAD (caspase-1 inhibitor) or when Caco-2 were incubated with anakinra, while butyrate incubation did not prevent such decrease. Taken together, early enteric α-synuclein accumulation contributes to compromise IEB through the direct activation of canonical caspase-1-dependent inflammasome signaling. These changes could contribute both to bowel symptoms as well as central pathology.

Mechano-electrical feedback in the clinical setting: Current perspectives.

Mechano-electric feedback (MEF) is an established mechanism whereby myocardial deformation causes changes in cardiac electrophysiological parameters. Extensive animal, laboratory and theoretical investigation has demonstrated that abnormal patterns of cardiac strain can induce alteration of electrical excitation and recovery through MEF, which can potentially contribute to the establishment of dangerous arrhythmias. However, the clinical relevance of MEF in patients with heart disease remains to be established. This paper reviews up-to date experimental evidence describing the response to different types of mechanical stimuli in the intact human heart with the support of new data collected during cardiac surgery. It discusses modulatory effects of MEF that may contribute to increase the vulnerability to arrhythmia and describes MEF interaction with clinical conditions where mechanically induced changes in cardiac electrophysiology are likely to be more relevant. Finally, directions for future studies, including the need for in-vivo human data providing simultaneous assessment of the distribution of structural, functional and electrophysiological parameters at the regional level, are identified.

1,2,3-Triazole Bridge as Conformational Constrain in ß-Hairpin Peptides: Analysis of Hydrogen-Bonded Positions.

Conformational constrained ß-hairpin peptides are useful tool to modulate protein-protein interactions. A triazole bridge in hydrogen-bonded positions between two antiparallel strands induces a conformational stabilization of the ß-hairpin peptide. The entity of the stability of the ß-hairpin peptide depends on the length of the bridge.

Hepatic left lobe volume is a sensitive index of metabolic improvement in obese women after gastric banding.

BACKGROUND: Nonalcoholic fatty liver disease is a common finding in obese subjects. Increasing evidence has been provided suggesting that it represents the hepatic component of the metabolic syndrome. OBJECTIVE: Aim of this longitudinal study was to evaluate the relationships between several anthropometric measures, including the hepatic left lobe volume (HLLV), and various indicators of the metabolic syndrome in a cohort of severely obese women before and after laparoscopic adjustable gastric banding (LAGB). STUDY DESIGN AND RESULTS: Seventy-five obese women (mean age 45 ± 10 years and body mass index (BMI) 42.5 ± 4.8 kg m(-2)) underwent LAGB and completed an average (± s.d.) post-surgical follow-up of 24 ± 6 months. Determination of HLLV, subcutaneous and intra-abdominal fat (IAF) was based on ultrasound. The principal component statistical analysis applied to pre-operative measurements, highlighted HLLV as a parameter that clustered with serum insulin, IAF, serum glucose and uric acid, along with triglycerides (TGs), alkaline phosphatase and high-density lipoprotein cholesterol. After LAGB, the average reduction of BMI was 23%, 12% for subcutaneous fat (SCF), 42% for HLLV and 40% for visceral fat. Among body weight, BMI, SCF, IAF and HLLV, reduction of the latter was an independent predictor of reduction of serum transaminases and γ-Glutamyltransferase, glucose, insulin and TGs. CONCLUSIONS: In severely obese women: (i) HLLV is a sensitive indicator of ectopic fat deposition, clustering with parameters defining the metabolic syndrome; (ii) weight loss achieved by LAGB is associated with a reduction of liver volume as estimated by HLLV; (iii) among various anthropometric parameters measured, reduction of HLLV that follows LAGB represents the best single predictor of improvement of various cardiometabolic risk factors.

Remote ischaemic preconditioning reduces myocardial injury in patients undergoing cardiac surgery with cold-blood cardioplegia: a randomised controlled trial.

BACKGROUND: Remote ischaemic preconditioning (RIPC) induced by brief ischaemia and reperfusion of the arm reduces myocardial injury in coronary artery bypass (CABG) surgery patients receiving predominantly cross-clamp fibrillation for myocardial protection. However, cold-blood cardioplegia is the more commonly used method world wide. OBJECTIVE: To assess whether RIPC is cardioprotective in CABG patients receiving cold-blood cardioplegia. DESIGN: Single-centre, single-blinded, randomised controlled trial. SETTING: Tertiary referral hospital in London. PATIENTS: Adults patients (18-80 years) undergoing elective CABG surgery with or without concomitant aortic valve surgery with cold-blood cardioplegia. Patients with diabetes, renal failure (serum creatinine >130 mmol/l), hepatic or pulmonary disease, unstable angina or myocardial infarction within the past 4 weeks were excluded. INTERVENTIONS: Patients were randomised to receive either RIPC (n = 23) or control (n = 22) after anaesthesia. RIPC comprised three 5 min cycles of right forearm ischaemia, induced by inflating a blood pressure cuff on the upper arm to 200 mm Hg, with an intervening 5 min reperfusion. The control group had a deflated cuff placed on the upper arm for 30 min. MAIN OUTCOME MEASURES: Serum troponin T was measured preoperatively and at 6, 12, 24, 48 and 72 h after surgery and the area under the curve (AUC at 72 h) calculated. RESULTS: RIPC reduced absolute serum troponin T release by 42.4% (mean (SD) AUC at 72 h: 31.53 (24.04) microg/l.72 h in controls vs 18.16 (6.67) microg/l.72 h in RIPC; 95% CI 2.4 to 24.3; p = 0.019). CONCLUSIONS: Remote ischaemic preconditioning induced by brief ischaemia and reperfusion of the arm reduces myocardial injury in CABG surgery patients undergoing cold-blood cardioplegia, making this non-invasive cardioprotective technique widely applicable clinically. TRIAL REGISTRATION NUMBER: NCT00397163.

Erythropoietin protects the human myocardium against hypoxia/reoxygenation injury via phosphatidylinositol-3 kinase and ERK1/2 activation.

BACKGROUND AND PURPOSES: Erythropoietin (EPO) has been shown to protect against myocardial infarction in animal studies by activating phosphatidylinositol-3 kinase (PI3K)/Akt and ERK1/2. However these pro-survival pathways are impaired in the diabetic heart. We investigated the ability of EPO to protect human atrial trabeculae from non-diabetic and diabetic patients undergoing coronary artery bypass surgery, against hypoxia-reoxygenation injury. EXPERIMENTAL APPROACH: Human atrial trabeculae were exposed to 90min hypoxia and 120min reoxygenation. EPO was administered throughout reoxygenation. The developed force of contraction, calculated as a percentage of baseline force of contraction, was continuously monitored. The involvement of PI3K and ERK1/2 and the levels of activated caspase 3(AC3) were assessed. KEY RESULTS: EPO improved the force of contraction in tissue from non-diabetic patients (46.7+/-1.7% vs. 30.2+/-2.2% in control, p<0.001). These beneficial effects were prevented by the PI3K inhibitor, LY294002 and the ERK1/2 inhibitor, U0126. EPO also significantly improved the force of contraction in the diabetic tissue, although to a lesser degree. The levels of activated caspase 3 were significantly reduced in EPO treated trabeculae from both non-diabetic and diabetic patients, relative to their respective untreated controls. CONCLUSIONS AND IMPLICATIONS: EPO administered at reoxygenation protected human myocardial muscle by activating PI3K and ERK1/2 and reducing the level of activated caspase 3. This cardioprotection was also observed in the diabetic group. This data supports the potential of EPO being used as a novel cardioprotective strategy either alone or as an adjunct in the clinical setting alongside existing reperfusion therapies.

Purification and properties of ornithine carbamoyltransferase from loggerhead turtle liver.

Ornithine carbamoyltransferase has been purified from the liver of the loggerhead turtle Caretta caretta by a single-step procedure using chromatography on an affinity column to which the transition-state analogue, delta-N-(phosphonoacetyl)-L-ornithine (delta-PALO), was covalently bound. The procedure employed yielded an enzyme which was purified 373-fold and was judged to be homogeneous by nondenaturing and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme showed a specific activity of 224. The molar mass of the C. caretta enzyme was approximately 112 kDa, the single band obtained by SDS-PAGE indicated a subunit molar mass of 39.5 kDa; hence, the enzyme is a trimer of identical subunits. It catalyzes an ordered sequential mechanism in which carbamoyl phosphate binds first, followed by L-ornithine. The Michaelis constants were 0.858 mM for L-ornithine and 0.22 mM for carbamoyl phosphate, the dissociation constant of the enzyme-carbamoyl phosphate complex was 0.50 mM.

Early single centre experience with 192 Sorin Bicarbon valves.

In 1990 Sorin Biomedica introduced a new pyrolytic bileaflet heart valve, the Bicarbon, developed using innovative design concepts with new materials, housing, hinge mechanism, leaflet curvature and sewing ring construction. From June 1991 to May 1995, 192 bicarbon valves were implanted in 177 patients by one team. The patient population was 102 males and 75 females with an age range from 16 to 82 years (mean 62.5 years). Ninety-six were AVR, 68 MVR, 12 double valve replacements, 2 patients received another type of bileaflet valve as did the triple valve replacement. Overall hospital mortality was 5.2%. For MVR the mortality was 12.7%; for AVR it was 4.7%. There were no deaths associated with double or triple valve replacement. Five deaths were at reoperation, 2 at emergency operation, 1 aortic dissection, 2 for endocarditis, none of the deaths were valve related. One patient had a haemorrhage due to inadequate control of Warfarin therapy. Four valves were explanted: 3 mitrals for paravalvular leaks and 1 aortic for endocarditis. No patients showed any evidence of thromboembolic complications and there were no late deaths. All the surviving patients have improved their preoperative clinical status. Maximum follow-up is nearly 4 years, mean 32 months (range 1-47). Twenty one patients underwent haemodynamic evaluation by Doppler echocardiography. The mean pressure drop across 12 mitral valves was 8.34 mmHg (mean size 29.7 mm) and for 9 aortic valves it was 15.85 mmHg (mean size 23). Based on these results we are of the opinion that the Bicarbon valve is equivalent to the best of other bileaflet valves currently available.

One hundred patients' experience with the Jyros valve.

The ideal mechanical valve abolishes all potential areas for blood stagnation and hence the potential for micro-emboli and clot formation. This problem has been addressed by the innovative design of the Jyros valve which has evolved from the technology available from Russian space research. The valve comprises two pyrolytic carbon leaflets which can rotate within the solid carbon housing to either align themselves to the optimum haemodynamic configuration, or to respond by rotation either continuous or intermittent, according to the degree of swirl on the inflow profile. This Jyros mechanical heart valve has been assessed in vivo at the London Chest Hospital since August 1992 where the series of unselected patients constitutes the largest single user experience available. 107 valves were implanted in 100 patients of age range 33 to 80 years. 23 patients were re-replacement for failed xenograft and 26 underwent concomitant revascularisation. 92 patients left hospital and 4 died later all from non-valve related causes. Two valves were explanted for endocarditis (one pre-existing) but successfully re-replaced. There have been no late thrombo-embolic events. Two patients with AVR & CABG had strokes at 8 months. There is no excessive haemolysis in excess of that detected for any mechanical valve nor any mechanical valve-related failure. Rotation of the valve is variable and can be visualised by 2D and M mode echo and by X-ray screening as the leaflets are impregnated with Boron carbide. Half of the valves rotate constantly and a quarter intermittently. There appears to be no disadvantage to patients in whom no valve rotation could be demonstrated. We have found that the theoretical potential of performance have been achieved.

Does activated neutrophil depletion on bypass by leukocyte filtration reduce myocardial damage? A preliminary report.

An important factor in the production of myocardial damage following cardiopulmonary bypass in the creation of oxygen derived free radicals. Few sources for these radicals have been identified but experimentally activated neutrophils are known to release free radical which contribute to myocyte necrosis. The aim of this pilot study was to identify whether, by depleting patients of leukocytes and particularly neutrophils on bypass, a better degree of myocardial protection could be observed using specific identifiers of myocardial damage. Ten patients undergoing urgent coronary artery bypass for unstable angina with impaired left ventricular function were leuko-depleted using a PALL medical leukocyte filter in the extra corporeal circulation together with leukocyte depletion of all transfused blood. A similar group of matched controls had only an arterial line filter without leukodepletion. All patients were operated by one surgeon using identical techniques of intermittent cross clamping and fibrillation at moderate hypothermia. Full blood count, Glutathione, Troponin T and CPK/MB were measured before, during and at identified intervals up to 72 hour after bypass. Preliminary results show little change in the total leukocyte count but the Troponin T and CPK/MB values were lower in the filtered group than in the control group and an increased level of total Glutathione in the filter group showed that there was less oxidated stress on the myocardium. Currently this filter is an expensive addition to bypass surgery but these preliminary results suggest that activated neutrophil depletion on bypass may be of benefit to patients with unstable angina, impending myocardial necrosis and low ejection fraction.

An essential lysine in the substrate-binding site of ornithine carbamoyltransferase.

Treatment of ornithine carbamoyltransferase from dolphin Stenella with pyridoxal phosphate, followed by reduction with NaBH4 resulted in complete loss of enzyme activity. The phosphate alone or the substrate analogue 2-aminovaleric acid moderately decreased the extent of inactivation, while carbamoyl phosphate plus 2-aminovaleric acid provided complete protection from inactivation. The partially inactivated enzyme showed K(m) values for substrates equivalent to those of native enzyme and lowered Kcat values. Two lysyl residues were substantially modified in the absence of ligands but only one of them was responsible for the inactivation of catalytic activity. Modification of a single subunit was sufficient to completely abolish the catalytic activity of the trimeric enzyme. The lysine involved has been identified as lysine 56 on the known primary structure of homologous human liver enzyme.

Cytotoxic and mutagenic effects of radon and radon daughters on murine L5178Y lines differing in DNA repair.

The effects of 222Rn were measured in mouse L5178Y (LY) lymphoblasts that differ in repair capabilities. Line LY-S1 is deficient in the repair of X-radiation-induced DNA doublestrand breaks, while lines LY-R16 and LY-R83 are presumed to be deficient in the excision of UV-radiation-induced pyrimidine dimers. Line LY-R83 is hemizygous while the other two lines are heterozygous at the thymidine kinase (tk) locus. After exposure to radon the D0's were found to be very similar for the three lines (0.30-0.31 Gy), whereas for X radiation the D0 for line LY-S1 is lower (0.7 Gy) than that for the two LY-R lines (1.3 Gy). Mutant frequencies at the tk locus were higher per gray after treatment with radon than X radiation, but at equitoxic doses the mutant frequencies were similar for X and alpha-particle radiation. A low radon-induced mutant frequency was observed for the hemizygous line, in agreement with the hypothesis that multilocus lesions were induced by the alpha-particle radiation and that mutants bearing intergenic lesions were not recovered in the TK+/- line. The entire active tk allele was lost by 81% of the TK-/- mutants of line LY-R16. In lines LY-S1 and LY-R16, 39-43% of the TK-/- mutants exhibited loss of galactokinase activity, indicating that the mutational lesion inactivating the tk gene frequently extended to the neighboring galactokinase gene.

Induction of multilocus lesions by UVC-radiation in mouse L5178Y lymphoblasts.

The survival, the mutant frequency and the nature of the DNA alteration responsible for the inactivation of the thymidine kinase (tk) locus were investigated in 5 strains of mouse L5178Y lymphoblasts exposed to UVC radiation. The nature of the DNA alteration was investigated in independent TK-/- mutants using Southern blot analysis. The concomitant loss of galactokinase (GK) activity in homogenates of individual TK-/- mutants was taken as an indication that the lesion inactivating the tk allele extended to the neighboring galactokinase (gk) allele. The survival of strains LY-R16 and LY-R83 was decreased to a greater extent than that of strains LY-S1, LY-SR1, and LY-3.7.2C, reflecting a deficiency in excision repair in strains derived from LY-R cells. The TK-/- mutant frequency of strain LY-R83, which is monosomic for chromosome 11 and thus hemizygous for the tk and gk genes, was only 50% of the mutant frequency of strain LY-R16 which is heterozygous for the tk gene. Moreover, a greatly reduced percentage of individual spontaneous and UVC-induced TK-/- mutants of strain LY-R83 showed loss of GK activity in comparison to the other strains. This result indicates that UVC irradiation induces intergenic mutations and that such mutants are poorly recovered in the hemizygous strain. Strain LY-3.7.2C appears to have only one active galactokinase (gk) allele, and very few TK-/- mutants of this strain showed loss of GK activity, possibly because this strain, although heterozyogous for the tk gene, is hemizygous in the region of the gk gene. Strains LY-R16 and LY-S1 are deficient in the repair of UVC- and X-radiation-induced damage, respectively, and the percentage of TK-/- mutants with intergenic mutations was higher for strain LY-R16 after UVC-radiation and for strain LY-S1 after X-radiation. These results indicate that unrepaired DNA lesions lead to an increase in intergenic mutations.

[Angioimmunoblastic lymphadenopathy. Treatment of 2 cases]. / La linfoadenopatia angioimmunoblastica. Osservazione di 2 casi.

Following a review of the literature, 2 cases of angioimmunoblastic lymphadenopathy with dysproteinaemia are reported. Anatomo-pathological examination of the lymphonodes shows widespread change in the structure with vascular and lympho-immunoblastic proliferation and deposit of amorphous eosinophil substance. The most significant clinical and morphological aspects are reported and these are related to the therapeutic response. The effectiveness of cortisone therapy is shown by comparison of laboratory examination data at admittance and after three months.