Hydrogen sulfide dysfunction in metabolic syndrome-associated vascular complications involves cGMP regulation through soluble guanylyl cyclase persulfidation.

Here, by using in vitro and ex vivo approaches, we elucidate the impairment of the hydrogen sulfide (H2S) pathway in vascular complications associated with metabolic syndrome (MetS). In the in vitro model simulating hyperlipidemic/hyperglycemic conditions, we observe significant hallmarks of endothelial dysfunction, including eNOS/NO signaling impairment, ROS overproduction, and a reduction in CSE-derived H2S. Transitioning to an ex vivo model using db/db mice, a genetic MetS model, we identify a downregulation of CBS and CSE expression in aorta, coupled with a diminished L-cysteine-induced vasorelaxation. Molecular mechanisms of eNOS/NO signaling impairment, dissected using pharmacological and molecular approaches, indicate an altered eNOS/Cav-1 ratio, along with reduced Ach- and Iso-induced vasorelaxation and increased L-NIO-induced contraction. In vivo treatment with the H2S donor Erucin ameliorates vascular dysfunction observed in db/db mice without impacting eNOS, further highlighting a specific action on smooth muscle component rather than the endothelium. Analyzing the NO signaling pathway in db/db mice aortas, reduced cGMP levels were detected, implicating a defective sGC/cGMP signaling. In vivo Erucin administration restores cGMP content. This beneficial effect involves an increased sGC activity, due to enzyme persulfidation observed in sGC overexpressed cells, coupled with PDE5 inhibition. In conclusion, our study demonstrates a pivotal role of reduced cGMP levels in impaired vasorelaxation in a murine model of MetS involving an impairment of both H2S and NO signaling. Exogenous H2S supplementation through Erucin represents a promising alternative in MetS therapy, targeting smooth muscle cells and supporting the importance of lifestyle and nutrition in managing MetS.

Duchenne's muscular dystrophy involves a defective transsulfuration pathway activity.

Duchenne muscular dystrophy (DMD) is the most frequent X chromosome-linked disease caused by mutations in the gene encoding for dystrophin, leading to progressive and unstoppable degeneration of skeletal muscle tissues. Despite recent advances in the understanding of the molecular processes involved in the pathogenesis of DMD, there is still no cure. In this study, we aim at investigating the potential involvement of the transsulfuration pathway (TSP), and its by-end product namely hydrogen sulfide (H2S), in primary human myoblasts isolated from DMD donors and skeletal muscles of dystrophic (mdx) mice. In myoblasts of DMD donors, we demonstrate that the expression of key genes regulating the H2S production and TSP activity, including cystathionine γ lyase (CSE), cystathionine beta-synthase (CBS), 3 mercaptopyruvate sulfurtransferase (3-MST), cysteine dioxygenase (CDO), cysteine sulfonic acid decarboxylase (CSAD), glutathione synthase (GS) and γ -glutamylcysteine synthetase (γ-GCS) is reduced. Starting from these findings, using Nuclear Magnetic Resonance (NMR) and quantitative Polymerase Chain Reaction (qPCR) we show that the levels of TSP-related metabolites such as methionine, glycine, glutathione, glutamate and taurine, as well as the expression levels of the aforementioned TSP related genes, are significantly reduced in skeletal muscles of mdx mice compared to healthy controls, at both an early (7 weeks) and overt (17 weeks) stage of the disease. Importantly, the treatment with sodium hydrosulfide (NaHS), a commonly used H2S donor, fully recovers the impaired locomotor activity in both 7 and 17 old mdx mice. This is an effect attributable to the reduced expression of pro-inflammatory markers and restoration of autophagy in skeletal muscle tissues. In conclusion, our study uncovers a defective TSP pathway activity in DMD and highlights the role of H2S-donors for novel and safe adjuvant therapy to treat symptoms of DMD.

Is hyaluronate sodium effective in the management of knee osteoarthritis? A placebo-controlled double-blind study.

AIM: The aim of this study was to investigate the effect of intra-articular hyaluronic acid (HA) injection on pain and function in knee osteoarthritis (OA). METHODS: Fourty-eight patients with knee OA were included in this study. The patients were randomized into two groups: one group received HA injections (average molecular weight [MW] 1.5 million Da), and the other group received placebo containing 0.9% saline. Three injections of HA or placebo were given at weeks 1, 2 and 3. The evaluation instruments were: Visual Analog Scale (VAS); Likert Scale; Lequesne 0.05), except for WOMAC pain subscore on walking at final assessment (week 14) which showed greater improvement in the HA-treated group (35.2% versus 9.1%; P=0.01). CONCLUSION: HA treatment was effective in the management of knee OA and improved knee pain and functional outcome, but there was no statistically significant difference in functional and symptom improvement with respect to saline (placebo) injection.

A family with autosomal dominant leukodystrophy linked to 5q23.2-q23.3 without lamin B1 mutations.

BACKGROUND AND PURPOSE: Duplications of lamin B1 (LMNB1) at 5q23 are implicated in adult-onset autosomal dominant leukodystrophy (ADLD) having been described in six families with diverse ethnic background but with a homogeneous phenotype. In a large Italian family, we recently identified a variant form of ADLD characterized clinically by absence of the autonomic dysfunction at onset described in ADLD and, on MRI, by milder cerebellar involvement with sparing of hemispheric white matter. Aim of this study was to investigate the genetic basis of this variant form of ADLD. METHODS: We carried out a genome-wide linkage analysis using microsatellite markers, and the genes in the candidate region were screened for point mutations. LMNB1 was also screened for deletions/duplications by real-time PCR, multiplex ligation-dependent probe amplification and Southern blot. RESULTS: We mapped the variant ADLD locus to 5q23.2-q23.3, a genomic region containing 11 genes including LMNB1. Neither gene copy-number defects nor point mutations in the LMNB1 gene were found. We also excluded point mutations in the coding exons of the other ten genes in the candidate region. However, expression of lamin B1 evaluated in lymphoblastoid cells was higher in patients than in healthy controls, and was similar to the lamin B1 expression levels found in a patient with LMNB1 duplication. CONCLUSIONS: This observation suggests that a mutation in an LMNB1 regulatory sequence underlies the variant ADLD phenotype. Thus, adult forms of ADLD linked to 5q23 appear to be more heterogeneous clinically and genetically than previously thought.

Autosomal recessive hereditary spastic paraplegia with thin corpus callosum: a novel mutation in the SPG11 gene and further evidence for genetic heterogeneity.

BACKGROUND AND PURPOSE: Autosomal Recessive Hereditary Spastic Paraplegia with Thin Corpus Callosum (AR-HSPTCC) is a clinically and genetically heterogeneous complicated form of spastic paraplegia. Two AR-HSPTCC loci have been assigned to chromosome 15q13-15 (SPG11) and chromosome 8p12-p11.21 respectively. Mutations in the SPG11 gene, encoding the spatacsin protein, have been found in the majority of SPG11 families. In this study, involvement of the SPG11 or 8p12-p11.21 loci was investigated in five Italian families, of which four consanguineous. METHODS: Families were tested for linkage to the SPG11 or 8p12-p11.21 loci and the SPG11 gene was screened in all the affected individuals. RESULTS: Linkage was excluded in the four consanguineous families. In the only SPG11-linked family the same homozygous haplotype 4.2 cM across the SPG11 locus was shared by all the three affected siblings. A novel c.2608A>G mutation predicted to affect the splicing was found in exon 14 of the SPG11 gene. DISCUSSION: This collection of families contributes to highlight the intra and inter locus heterogeneity in AR-HSPTCC, already remarked in previous reports. In particular, it confirms heterogeneity amongst Italian families and reports a new mutation predicted to affect splicing in the spatacsin gene.

Refinement of the SPG9 locus on chromosome 10q23.3-24.2 and exclusion of candidate genes.

BACKGROUND AND PURPOSE: The hereditary spastic paraplegias (HSPs) are a heterogeneous group of neurodegenerative disorders, characterized by a progressive spasticity of the lower limbs. So far, 33 different loci (SPGs) have been mapped and the 15 genes responsible have been identified. We mapped a locus responsible for a form of spastic paraplegia, complicated by bilateral cataracts, gastroesophageal reflux with persisting vomiting and amyotrophy to chromosome 10q23.3-q24.2, in an Italian family. The critical region was in a 12 cm chromosomal interval between markers D10S564 and D10S603 (SPG9, MIM601162). In the same region, two other forms of HSP have been recently mapped: SPG27 and SPG33. In the latter case, the gene responsible has been identified. MATERIALS AND METHODS: To better characterize this region, we genotyped individuals from SPG9-linked families using additional markers and reduced the candidate region to a 4.8 Mb, excluding several genes by positional cloning. RESULTS: The refined SPG9 locus is positioned completely within SPG27 and does not include the SPG33 gene. DISCUSSION: Fifty-two transcripts are present in the refined critical region and 25 strong candidates have been excluded as disease causing genes by direct sequencing. Six of them were also excluded as responsible for SPG27.

Depression and psychosocial factors in Turkish women with fibromyalgia syndrome.

AIM: The aim of this study was to evaluate the demographic qualifications, frequency of depression, degree of pain, and the correlations between these factors in Turkish women with fibromyalgia syndrome (FS). METHODS: Fifty-three women with FS and 54 healthy women were included in the study. The visual analogue scale (VAS) was applied to evaluate the degree of pain and Beck depression inventory (BDI) for depression. RESULTS: Mean age of the FS and the healthy women groups was 42.6+/-9.6 (21-63) and 39.4+/-13.2 (24-60) years respectively; which were statistically similar (P>0.05). Of the patients, 69.8% were married, and the mean years of education was 8.7+/-4.8 years. Marriage among relatives was found in 18.4% of the patients. The mean VAS score of the patients was 7.2+/-1.7 cm with the mean duration of pain 5+/-4.6 years. Mean BDI scores of FS patients and the healthy group were 15.7+/-8.7 and 10.2+/-5.5 respectively; the difference between the 2 groups was statistically significant (P<0.05). According to the BDI scores, 90% of FS patients were classified as depressed; among them, 50% had minor, 38% moderate, and 2% severe depression. FS patients who were married to a relative had higher scores of BDI (r=0.414, P=0.013). There was negative correlation between BDI score and patients' total year of education (r=-0.295, P=0.037); and the husband's education level (r=-0.367, P=0.030). According to BDI, the c2 test revealed significant depression in patients with sleep-disorders (P=0.009). CONCLUSIONS: We found a significant degree of depression in Turkish female FS patients. This situation is found to be correlated with the education level of both patient and husband; marital status, and sleep-disorder; it is suggested that these factors should be taken into consideration in the diagnosis, treatment and follow-up of FS patients.

Nonspecific inflammatory bowel disease and smoking.

The authors assessed the relation between cigarette smoking and nonspecific inflammatory bowel disease in a case-control study of 124 cases of ulcerative colitis, 109 cases of Crohn's disease, and 250 age- and sex-matched control subjects in hospital for acute nongastric or intestinal conditions unrelated to smoking. For ulcerative colitis, the risk for current smoking compared with never smoking was 0.5, with a 95% confidence interval (Cl) of 0.3-1.0. They observed decreasing risk with increasing number of cigarettes smoked. The risk for ex-smokers, however, was greater than that for never smokers (relative risk = 2.7; 95% Cl = 1.5-4.9). The elevated risk of ulcerative colitis in ex-smoking in the presence of an overall lack of association with ever-smoking may plausibly be attributed to either 1) brief induction time of a protective effect of smoking on ulcerative colitis or 2) selective cessation of smoking due perhaps to very early symptoms of the disease. If time at first onset of bowel symptoms, instead of clinical diagnosis, is considered as the index date, the negative association between ulcerative colitis and current smoking would have weakened in men and disappeared in the overall series. There was clear evidence of a positive association between cigarette smoking and Crohn's disease (relative risk for ever smokers vs. never smokers = 4.0; 95% Cl = 2.2-7.3). The risk estimates increased with the number of cigarettes smoked per day and duration of habit. The association between current smoking and Crohn's disease was even stronger when age at first onset of bowel symptoms was considered as the index date, but the risk for ex-smokers fell below unity.

Medium-dose antacids versus cimetidine in the short-term treatment of duodenal ulcer.

Seventy-eight patients with endoscopically proven duodenal ulcer were randomly allocated to be treated with a medium dose of liquid aluminum-magnesium antacid (75 ml in five daily doses) or cimetidine (400 mg twice daily) for 4 weeks in a prospective double-blind, double-dummy study. Healing rates at completion of trial were 66.7% in the cimetidine-treated group and 71.8% in the antacid group (p, ns). Both treatments were equally effective in relieving ulcer symptoms. Among the patient variables considered, only cigarette smoking was found to have a significant negative effect on ulcer healing. These results indicate that medium doses of antacids are as effective as cimetidine in the short-term treatment of duodenal ulcer.

A double-blind controlled trial of ranitidine 300 mg nocte and ranitidine 150 mg b.i.d. in the short-term treatment of gastric ulcer.

A multi-center study was carried out to compare the healing rates of ranitidine 300 mg nocte and 150 mg b.i.d. in the short-term treatment of gastric ulcers and to assess the side-effect liability of ranitidine 300 mg given in a single daily dose. Forty-five outpatients suffering from endoscopically and bioptically proven uncomplicated benign gastric ulcer were selected for the study. The patients were treated, for 4 weeks, on the basis of a double-blind randomized design. An endoscopic examination was repeated within 4 days after the end of the treatments. The patients who did not demonstrate complete healing were treated for an additional 4-week period. Clinical controls were performed to evaluate symptoms, antacid consumption, compliance with trial tablet consumption; hematological and biochemical tests were also carried out at the end of the 4 and 8-week periods. No differences were observed between the healing rate induced by 300 mg nocte and 150 mg b.i.d., after 4 weeks of treatment (76% and 79% respectively) and after 8 weeks (100% in both groups). Similar results were obtained as far as symptoms, antacid consumption and compliance are concerned. Neither treatment regimen induced appreciable side-effects.

The use of cimetropii bromidum as premedication for endoscopy of the upper gastro-intestinal tract: a double-blind controlled clinical trial.

The effectiveness of cimetropii bromidum (10 or 20 mg i.v.) as premedication for upper gastro-intestinal tract endoscopy was examined and compared with that of placebo in a double-blind crossover study involving 160 patients. The drug effects were assessed from the degree of opening of the pyloric sphincter and the endoscopist's rating of drug performance. The study showed that the effects of cimetropii bromidum differed highly significantly (p less than 0.0001) from those of placebo, on either count. No adverse effects were noted.