Prevalence of metabolic syndrome and non-alcoholic fatty liver disease in a cohort of italian patients with spinal-bulbar muscular atrophy.

Spinal-bulbar muscular atrophy (SBMA), is an X-linked motor neuron disease caused by a CAG-repeat expansion in the first exon of the androgen receptor gene (AR) on chromosome X. In SBMA, non-neural clinical phenotype includes disorders of glucose and lipid metabolism. We investigated the prevalence of metabolic syndrome (MS), insulin resistance (IR) and non alcoholic fatty liver disease (NAFLD) in a group of SBMA patients. Forty-seven consecutive patients genetically diagnosed with SBMA underwent biochemical analyses. In 24 patients abdominal sonography examination was performed. Twenty-three (49%) patients had fasting glucose above reference values and 31 (66%) patients had a homeostatic model assessment (HOMA-IR) ≥ 2.6. High levels of total cholesterol were found in 24 (51%) patients, of LDL-cholesterol in 18 (38%) and of triglycerides in 18 (38%). HDL-cholesterol was decreased in 36 (77%) patients. Twenty-four (55%) subjects had 3 or more criteria of MS. A positive correlation (r = 0.52; p < 0.01) was observed between HOMA-IR and AR-CAG repeat length. AST and ALT were above the reference values respectively in 29 (62%) and 18 (38%) patients. At ultrasound examination increased liver echogenicity was found in 22 patients (92 %). In one patient liver cirrhosis was diagnosed. Liver/kidney ratio of grey-scale intensity, a semi-quantitative parameter of severity of steatosis, strongly correlated with BMI (r = 0.68; p < 0.005). Our study shows a high prevalence of IR, MS and NAFLD in SBMA patients, conditions that increase the cardiovascular risk and can lead to serious liver damage, warranting pharmacological and non-pharmacological treatment.

A strain of Meyerozyma guilliermondii isolated from sugarcane juice is able to grow and ferment pentoses in synthetic and bagasse hydrolysate media.

The search for new microbial strains that are able to withstand inhibitors released from hemicellulosic hydrolysis and are also still able to convert sugars in ethanol/xylitol is highly desirable. A yeast strain isolated from sugarcane juice and identified as Meyerozyma guilliermondii was evaluated for the ability to grow and ferment pentoses in synthetic media and in sugarcane bagasse hydrolysate. The yeast grew in xylose, arabinose and glucose at the same rate at an initial medium pH of 5.5. At pH 4.5, the yeast grew more slowly in arabinose. There was no sugar exhaustion within 60 h. At higher xylose concentrations with a higher initial cell concentration, sugar was exhausted within 96 h at pH 4.5. An increase of 350 % in biomass was obtained in detoxified hydrolysates, whereas supplementation with 3 g/L yeast extract increased biomass production by approximately 40 %. Ethanol and xylitol were produced more significantly in supplemented hydrolysates regardless of detoxification. Xylose consumption was enhanced in supplemented hydrolysates and arabinose was consumed only when xylose and glucose were no longer available. Supplementation had a greater impact on ethanol yield and productivity than detoxification; however, the product yields obtained in the present study are still much lower when compared to other yeast species in bagasse hydrolysate. By the other hand, the fermentation of both xylose and arabinose and capability of withstanding inhibitors are important characteristics of the strain assayed.

Yeast composition of sugar cane juice in relation to plant varieties and seasonality / Composição de leveduras do caldo em relação às variedades de cana e sazonalidade

In the production of the artisanal cachaça, a beverage obtained after distillation of the fermented sugar cane juice, natural starter ferment ("fermento caipira") is utilized, in which crushed corn, rice bran and citric fruit juice are added to sugar cane juice. The primary microbial source is the juice itself, and although the cachaça sensorial quality is recognized when this ferment is utilized, difficulties in the quality control due to the high level of contaminants and extensive preparation periods are reported. In this context, this work aimed the evaluation of the yeast composition and physico-chemical characteristics of the juice extracted from 10 sugar cane RB-varieties during the harvest season in an area under organic management, seeking for information to contribute to the varietal management allowing a faster and efficient ferment preparation. A significant decrease in the yeast numbers in the juice was observed when the maximal point of maturity was reached for the majority of the varieties. However, the proportion (%) of Saccharomyces increased with the cane maturity, recommending the early and medium maturity varieties (RB835054, RB835486, RB845210 and RB855156) to be utilized at the beginning of the harvest period for the ferment preparation, which could result in diminished preparation time and faster fermentation. The variety RB845210 is indicated because it also presented high reducing sugar and protein concentrations in the juice. The varietal management can facilitate the production and performance of the natural starter ferment, in order to contribute for the organic cachaça production.

Na produção artesanal de cachaça, bebida obtida através da destilação do caldo de cana-de-açúcar fermentado, tradicionalmente utiliza-se o fermento natural ou também chamado de caipira, resultado da mistura de vários ingredientes como milho moído, farelo de arroz e suco de frutas cítricas com caldo de cana. A fonte primária de microrganismos é o próprio caldo da cana, e embora se reconheça a qualidade sensorial da bebida quando este tipo de fermento é utilizado, há alguns inconvenientes como dificuldades no controle de qualidade devido ao alto nível de contaminantes e longos períodos de preparação. Neste contexto, o objetivo deste trabalho foi avaliar a composição de leveduras e as características físico-químicas do caldo em relação às variedades de cana orgânica (10 variedades RB) e à sazonalidade, no intuito de gerar informações para o manejo de variedades que permita o preparo do fermento caipira de forma mais eficiente e rápida. Os resultados indicaram uma diminuição significativa no número de leveduras próximo ao ponto máximo de maturação da cana-de-açúcar para a maioria das variedades. Porém, observou-se que a proporção (%) de Saccharomyces aumentou em decorrência da maturação da cana. Sugere-se as variedades precoces e precoces/médias (RB835054, RB835486, RB845210 e RB855156) a serem utilizadas no início da safra para o preparo do fermento caipira, o que poderia proporcionar uma diminuição no tempo de preparo do fermento e uma fermentação mais rápida. A variedade RB845210 é indicada por apresentar também maior concentração de açúcar redutor e proteína no caldo. O manejo varietal pode facilitar a produção e eficiência do fermento caipira, contribuindo assim para a produção de cachaça orgânica.

Identification of four novel mutations in the alpha glucosidase gene in five Italian patients with infantile onset glycogen storage disease type II.

Glycogen storage disease type II (GSDII) is an autosomal recessive disorder due to the deficiency of the lysosomal enzyme acid alpha glucosidase. Four novel mutations (C670T, G989A, G2188T, and Delta 23 nt 828-850) were identified in five Italian patients with the infantile form of the disease. The C670T mutation was present in two unrelated patients in heterozygosity; the effect on enzyme activity was assessed by in vitro expression. COS-1 cells expressing the C670T allele had a twofold higher activity than the negative control cells. The G989A and G2188T point mutations lead to the introduction of premature stop signals that results in truncated forms of alpha glucosidase. The in vitro expression of G2188T allele demonstrated no increment in activity compared to negative control. The frame shifting deletion of nucleotides 828-850 was identified in one patient in heterozygosity. The shift in the reading frame introduces a stop codon 135 nucleotides downstream the deletion junction that results in a truncated protein without catalytic activity. Nested PCR screening showed that the mutation was carried by the mother and was absent in the other members of the family. The four novel severe mutations herein described concerned only infantile onset GSDII patients; the loss of enzyme activity is correlated with the severity of the disease.

Niemann-Pick type C disease: mutations of NPC1 gene and evidence of abnormal expression of some mutant alleles in fibroblasts.

We analyzed Niemann-Pick type C disease 1 (NPC1) gene in 12 patients with Niemann-Pick type C disease by sequencing both cDNA obtained from fibroblasts and genomic DNA. All the patients were compound heterozygotes. We found 15 mutations, eight of which previously unreported. The comparison of cDNA and genomic DNA revealed discrepancies in some subjects. In two unrelated patients carrying the same mutations (P474L and nt 2972del2) only one mutant allele (P474L), was expressed in fibroblasts. The mRNA corresponding to the other allele was not detected even in cells incubated with cycloheximide. The promoter variants (-1026T/G and -1186T/C or -238 C/G), found to be in linkage with 2972del2 allele do not explain the lack of expression of this allele, as they were also found in control subjects. In another patient, (N1156S/Q922X) the N1156S allele was expressed in fibroblasts while the expression of the other allele was hardly detectable. In a fourth patient cDNA analysis revealed a point mutation in exon 20 (P1007A) and a 56 nt deletion in exon 22 leading to a frameshift and a premature stop codon. The first mutation was confirmed in genomic DNA; the second turned out to be a T-->G transversion in exon 22, predicted to cause a missense mutation (V1141G). In fact, this transversion generates a donor splice site in exon 22, which causes an abnormal pre-mRNA splicing leading to a partial deletion of this exon. In some NPC patients, therefore, the comparison between cDNA and genomic DNA may reveal an unexpected expression of some mutant alleles of NPC1 gene.