altered vessel signalling molecules in subjects with down s syndrome

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Humoral and cellular response to BoHV-1 in buffalo and cattle treated with an inactivated marker vaccine

The study is aimed at assessing and comparing the immune response to BoHV-1 elicited by an inactivated marker vaccine in buffaloes and cattle. Vaccination did not produced any local or general reactions in buffaloes. Seroneutralizing antibodies and cellular response by IFN-γ- test have been detected in buffaloes and cattle after a prime/ booster vaccination strategy. Humoral and cellular responses were significantly higher in cattle than in buffaloes. Data pointed out the possibility to use the marker vaccine in buffaloes. However, further studies must be planned to assess the immune pressure of marker vaccines in terms of IBR eradicative attitude in infected buffalo herds

Reduced mitochondrial D-loop methylation levels in sporadic amyotrophic lateral sclerosis

Background: Mitochondrial dysregulation and aberrant epigenetic mechanisms have been frequently reported in neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS), and several researchers suggested that epigenetic dysregulation in mitochondrial DNA (mtDNA) could contribute to the neurodegenerative process. We recently screened families with mutations in the major ALS causative genes, namely C9orf72, SOD1, FUS, and TARDBP, observing reduced methylation levels of the mtDNA regulatory region (D-loop) only in peripheral lymphocytes of SOD1 carriers. However, until now no studies investigated the potential role of mtDNA methylation impairment in the sporadic form of ALS, which accounts for the majority of disease cases. The aim of the current study was to investigate the D-loop methylation levels and the mtDNA copy number in sporadic ALS patients and compare them to those observed in healthy controls and in familial ALS patients. Pyrosequencing analysis of D-loop methylation levels and quantitative analysis of mtDNA copy number were performed in peripheral white blood cells from 36 sporadic ALS patients, 51 age- and sex-matched controls, and 27 familial ALS patients with germinal mutations in SOD1 or C9orf72 that represent the major familial ALS forms. Results: In the total sample, D-loop methylation levels were significantly lower in ALS patients compared to controls, and a significant inverse correlation between D-loop methylation levels and the mtDNA copy number was observed. Stratification of ALS patients into different subtypes revealed that both SOD1-mutant and sporadic ALS patients showed lower D-loop methylation levels compared to controls, while C9orf72-ALS patients showed similar D-loop methylation levels than controls. In healthy controls, but not in ALS patients, D-loop methylation levels decreased with increasing age at sampling and were higher in males compared to females. Conclusions: Present data reveal altered D-loop methylation levels in sporadic ALS and confirm previous evidence of an inverse correlation between D-loop methylation levels and the mtDNA copy number, as well as differences among the major familial ALS subtypes. Overall, present results suggest that D-loop methylation and mitochondrial replication are strictly related to each other and could represent compensatory mechanisms to counteract mitochondrial impairment in sporadic and SOD1-related ALS forms

Curcuma longa Extract Exerts a Myorelaxant Effect on the Ileum and Colon in a Mouse Experimental Colitis Model, Independent of the Anti-Inflammatory Effect

Background: Curcuma has long been used as an anti-inflammatory agent in inflammatory bowel disease. Since gastrointestinal motility is impaired in inflammatory states, the aim of this work was to evaluate if Curcuma Longa had any Methods: The biological activity of Curcuma extract was evaluated against Carbachol induced contraction in isolated mice intestine. Acute and chronic colitis were induced in Balb/c mice by Dextran Sulphate Sodium administration (5% and 2.5% respectively) and either Curcuma extract (200 mg/kg/day) or placebo was thereafter administered for 7 and 21 days respectively. Spontaneous contractions and the response to Carbachol and Atropine of ileum and colon were studied after colitis induction and Curcuma administration. Results: Curcuma extract reduced the spontaneous contractions in the ileum and colon; the maximal response to Carbachol was inhibited in a non-competitive and reversible manner. Similar results were obtained in ileum and colon from Curcuma fed mice. DSS administration decreased the motility, mainly in the colon and Curcuma almost restored both the spontaneous contractions and the response to Carbachol after 14 days assumption, compared to standard diet, but a prolonged assumption of Curcuma decreased the spontaneous and Carbachol-induced contractions. Conclusions: Curcuma extract has a direct and indirect myorelaxant effect on mouse ileum and colon, independent of the anti-inflammatory effect. The indirect effect is reversible and non-competitive with the cholinergic agent. These results suggest the use of curcuma extract as a spasmolytic agent

Serum metal evaluation in a small cohort of Amyotrophic Lateral Sclerosis patients reveals high levels of thiophylic species

Amyotrophic Lateral Sclerosis (ALS) has often been associated with improper/altered metal metabolism. Analysis of thiophylic metals in serum from a small and geographically restricted cohort of ALS patients indicates contents of Pb and Ni much higher in patients than in controls (Ni, 5-fold; Pb, 2-fold). Se levels are also higher in the patients\u2019 group, which has instead lower As levels than controls. Thiophylic metals may impair biogenesis of FeS clusters or substitute for iron, even in folded proteins; Se may non-functionally replace S. Thus, improper assembly/function of FeS proteins could represent another possible issue to be considered in ALS pathogenesis

Metal and proteomic analysis of sporadic ALS patients with common geographical origin

Amyotrophic Lateral Sclerosis (ALS) is a rare neurodegenerative disorder characterized by selective degeneration of both upper and lower motor neurons in the brain, brainstem, and spinal cord. This results in paralysis due to muscle weakness and atrophy, leading to death in 3-5 years. Genetic and environmental factors are involved in the pathogenesis of the disease and metals metabolism have been linked to ALS. This study enrolled seven patients and five controls (age matched, living in the same geographical area). For metal quantitation, samples of serum were analyzed by ICP-MS. For proteomic analyses, immobilized pH gradient covered the 4-10 and 3-7 pH range. Statistical analyses were carried out with Student's t-test and Artificial Neural Networks. Among the metals analyzed, As concentration resulted significantly lower in patients than in controls (p=0.007); Hg too was found in lower concentration in patients, but with a lower statistical significance (p=0.13). Higher concentration of Al in patients was detected (p=0.08). In this study, we were not able to confirm the higher concentrations of Ni and Pb in patients previously described in a smaller cohort. Our proteomics data show that APOA2 is decreased by 30% in patients with respect to controls. Furthermore, AHSG and SAP showed a significant decrease in patients with a story of more than 10 years of disease. Impaired metal homeostasis, attributable to environmental exposure, could lead to mineral overload. Besides promoting oxidative stress, metals can compete for the binding sites of metal-containing proteins, such as those containing iron-sulfur clusters. At present, no literature data link APOA2 to ALS, but the fact that its mRNA is processed by TDP43, provides a possible connection with the disease. The proteins differentially expressed belong to the group of Acute Phase Reaction proteins, possibly linking ALS to a chronic inflammation status. Further experiments are still ongoing

A multidisciplinary approach to study Sporadic Amyotrophic Lateral Sclerosis in patients with common geographical origin

Amyotrophic Lateral Sclerosis (ALS), a fatal neurodegenerative disorder, is object of intensive research as the causes are still unknown and a treatment not available yet. This project is aimed to study, with a multidisciplinary approach, a small cohort of ALS subjects with a common environmental exposure. For metal quantitation, samples of serum and whole blood were analyzed by ICP-MS. For proteomic analyses, immobilized pH gradient covered the 4-10 and 3-7 pH range. Arsenic concentration resulted significantly lower in patients than in controls. Also, Mn and Hg showed lower levels in patients. Levels of plasma APOA2 protein resulted decreased in patients with respect to controls, whereas SAMP showed a significant decrease only in the late onset group. APOA1 and TTHY also were decreased, the latter in late-onset patients. RET4 was decreased only in the early-onset group. When evaluating APOE genotype we found a 3-fold increase in the frequency of E3/E4 genotype in the patient\u2019s group. DNA oxidative stress has been evaluated through a Comet Assay. The multidisciplinary approach applied in this study allowed to dissect different aspects of ALS, often are evaluated separately and in heterogeneous cohorts of patients

Metal and proteomic analysis of sporadic ALS patients with common geographical origin

Amyotrophic Lateral Sclerosis (ALS) is a rare neurodegenerative disorder characterized by selective degeneration of both upper and lower motor neurons in the brain, brainstem, and spinal cord. This results in paralysis due to muscle weakness and atrophy, leading to death in 3-5 years. Genetic and environmental factors are involved in the pathogenesis of the disease and metals metabolism have been linked to ALS. This study enrolled seven patients and five controls (age matched, living in the same geographical area). For metal quantitation, samples of serum were analyzed by ICP-MS. For proteomic analyses, immobilized pH gradient covered the 4-10 and 3-7 pH range. Statistical analyses were carried out with Student's t-test and Artificial Neural Networks. Among the metals analyzed, As concentration resulted significantly lower in patients than in controls (p=0.007); Hg too was found in lower concentration in patients, but with a lower statistical significance (p=0.13). Higher concentration of Al in patients was detected (p=0.08). In this study, we were not able to confirm the higher concentrations of Ni and Pb in patients previously described in a smaller cohort. Our proteomics data show that APOA2 is decreased by 30% in patients with respect to controls. Furthermore, AHSG and SAP showed a significant decrease in patients with a story of more than 10 years of disease. Impaired metal homeostasis, attributable to environmental exposure, could lead to mineral overload. Besides promoting oxidative stress, metals can compete for the binding sites of metal-containing proteins, such as those containing iron-sulfur clusters. At present, no literature data link APOA2 to ALS, but the fact that its mRNA is processed by TDP43, provides a possible connection with the disease. The proteins differentially expressed belong to the group of Acute Phase Reaction proteins, possibly linking ALS to a chronic inflammation status. Further experiments are still ongoing

Dissecting Sporadic ALS in a geographical cluster of patients: a multidisciplinary study

Amyotrophic Lateral Sclerosis (ALS) is a rare neurodegenerative disorder that results in paralysis and leading to death in 3-5 years. Genetic and environmental factors are involved in the pathogenesis of this disease and metals metabolism has been linked to ALS. Proteomic studies are currently being performed to search for possible biomarkers. Here we present a study aimed at investigating different aspects of the disease, based on a multidisciplinary approach. The cohort of ALS patients that we analyzed includes seven patients, all originating from a common, restricted, geographical area and five matched controls. Environmental exposure is the same for all these subjects. SOD1, FUS, TDP43, C9ORF72 and APOE genotypes were evaluated. For metal quantitation, samples of serum and whole blood were analyzed by ICP-MS. For proteomic analyses, immobilized pH gradient covered the 4-10 and 3-7 pH range both in reducing and nonreducing conditions. Levels of DNA oxidation were evaluated by a comet assay. Statistical analyses were carried out with Student\u2019s t-test and Artificial Neural Networks. As concentration resulted significantly lower in patients than in controls ( Auto-CM analysis linked closely high concentrations of Al and Se to the ALS group. Levels of metals in whole blood have been correlated with levels in serum. Proteomics data show that some proteins related to Acute Phase Response (APR) and lipid homeostasis are decreased in patients. Apo\u3b54 allele is more represented in the patient\u2019s group than in controls\u2019. Impaired metal homeostasis, attributable to environmental exposure, could lead to mineral overload. Metals can compete for the binding sites of metal-containing proteins. The different expression of the APR proteins reported could be a reflection of the disease status of the subjects analyzed. Enrichment in Apo\u3b54 allele frequency in patients may provide a link between neurodegeneration and lipid metabolism disturbances

BS148 Reduces the Aggressiveness of Metastatic Melanoma via Sigma-2 Receptor Targeting

: The management of advanced-stage melanoma is clinically challenging, mainly because of its resistance to the currently available therapies. Therefore, it is important to develop alternative therapeutic strategies. The sigma-2 receptor (S2R) is overexpressed in proliferating tumor cells and represents a promising vulnerability to target. Indeed, we have recently identified a potent S2R modulator (BS148) that is effective in melanoma. To elucidate its mechanism of action, we designed and synthesized a BS148 fluorescent probe that enters SK-MEL-2 melanoma cells as assessed using confocal microscopy analysis. We show that S2R knockdown significantly reduces the anti-proliferative effect induced by BS148 administration, indicating the engagement of S2R in BS148-mediated cytotoxicity. Interestingly, BS148 treatment showed similar molecular effects to S2R RNA interference-mediated knockdown. We demonstrate that BS148 administration activates the endoplasmic reticulum stress response through the upregulation of protein kinase R-like ER kinase (PERK), activating transcription factor 4 (ATF4) genes, and C/EBP homologous protein (CHOP). Furthermore, we show that BS148 treatment downregulates genes related to the cholesterol pathway and activates the MAPK signaling pathway. Finally, we translate our results into patient-derived xenograft (PDX) cells, proving that BS148 treatment reduces melanoma cell viability and migration. These results demonstrate that BS148 is able to inhibit metastatic melanoma cell proliferation and migration through its interaction with the S2R and confirm its role as a promising target to treat cancer