Draft Genome Sequence of Tsukamurella sp. Strain 1534

A draft genome sequence of Tsukamurella sp., an aerobic bacterium isolated from a human sputum specimen, is described here. A new virus or provirus, TPA4, was characterized.     

Association of three polymorphisms of scavenger receptor class BI gene (exon8, exon1, intron5) with coronary stenosis in a coronary Tunisian population

Background

The potential role of scavenger receptor class BI (gene name SCARB1) in the regulation of lipoproteins metabolism and atherosclerosis has attracted considerable interest. We tested the relationship of SCARB1 polymorphisms with significant coronary stenosis (SCS) and lipid profile in a coronary Tunisian population.

Methods

Three SCARB1 polymorphisms (exon8 (C/T), exon1 (G/A), intron5 (C/T)) were studied in 316 Tunisian patients undergoing coronary angiography. SCS was defined as a luminal narrowing of ≥ 50% in at least one major coronary artery. Lipid profile was measured. Genotyping was performed using PCR–RFLP.

Results

Individuals with TT genotypes of exon8 were associated with higher concentrations of plasma HDL-C and ApoAI in the group without SCS. Carriers of T allele of exon8 were associated with 41% lower risk of SCS. This protective effect seemed to be particularly significant in women, nondiabetics and nonsmokers. Subjects homozygous for the variant allele of intron5 were significantly associated with an increased risk of SCS, particularly in smokers. AA genotype of exon1 was associated with an increased risk of SCS in diabetics and in patients with metabolic syndrome. The (CAT) haplotype was associated with increase in the risk of SCS compared to the wild haplotype and had a 4-fold greater risk of SCS than patients with haplotype (TGC) which seems to be the most protective against SCS.

Conclusion

Carriers of T allele of exon8 in SCARB1 seemed to increase HDL-C and ApoAI concentrations and reduce the risk of SCS. The intron5, exon1 and (CAT) haplotype seemed to have an atherogenic effect.     

Development of a beverage from red grape juice fermented with the Kombucha consortium

Kombucha is a health-promoting fermented beverage traditionally made by fermenting a sweetened tea with a symbiotic culture of yeast species and acetic acid bacteria. The aim of this work was to develop a beverage using red grape juice as an alternative substrate. Grape juice contains various nutrient elements and phytochemicals, such as polyphenols, which possess a wide range of biological activities. We investigated the chemical characteristics and sensory and antimicrobial activities of the fermented grape juice Kombucha beverage. The pH decreased from 3.95 to 2.9 during the fermentation process and remained fairly constant thereafter, and the acetic acid bacteria and yeast counts in the broth increased up to 6 days of fermentation and subsequently decreased. Phenolic and anthocyanin contents and the antioxidant activity of the fermented beverage were higher after fermentation, with the maximum increase observed on the sixth day of fermentation when values were approximately 2.47- and 1.59-fold higher than pre-fermentation values, respectively, as assessed by 2,2-diphenyl-1-picrylhydrazyl and 2,2′-azino-bis (3-ethylbenzothiazoline-6- sulfonic acid) radical scavenging assays. Fourier transform infrared spectroscopy was used for the qualitative analysis of the grape juice before and after fermentation. Distinct peak variations in the spectral region between 2500 and 1650 cm^?1 were observed, which matched the appearance of organic acids and changes in phenolic compounds. Fermented juice Kombucha showed antibacterial activity toward all tested bacteria, which can be primarily ascribed to the increased production of acetic acid, but also to the biosynthesis of other metabolites, during the fermentation process. The 6-day fermented juice was the most appreciated by the taste panel based on the overall quality evaluation; with prolongation of fermentation the fermented juice acquired a distinct sour flavor.     

Early oxidative stress in the diabetic kidney: effect of DL-alpha-lipoic acid

Oxidative stress is implicated in the pathogenesis of diabetic nephropathy. The attempts to identify early markers of diabetes-induced renal oxidative injury resulted in contradictory findings. We characterized early oxidative stress in renal cortex of diabetic rats, and evaluated whether it can be prevented by the potent antioxidant, DL-alpha-lipoic acid. The experiments were performed on control rats and streptozotocin-diabetic rats treated with/without DL-alpha-lipoic acid (100 mg/kg i.p., for 3 weeks from induction of diabetes). Malondialdehyde plus 4-hydroxyalkenal concentration was increased in diabetic rats vs. controls (p <.01) and this increase was partially prevented by DL-alpha-lipoic acid. F(2) isoprostane concentrations (measured by GCMS) expressed per either mg protein or arachidonic acid content were not different in control and diabetic rats but were decreased several-fold with DL-alpha-lipoic acid treatment. Both GSH and ascorbate (AA) levels were decreased and GSSG/GSH and dehydroascorbate/AA ratios increased in diabetic rats vs. controls (p <.01 for all comparisons), and these changes were completely or partially (AA) prevented by DL-alpha-lipoic acid. Superoxide dismutase, glutathione peroxidase, glutathione reductase, glutathione transferase, and NADH oxidase, but not catalase, were upregulated in diabetic rats vs. controls, and these activities, except glutathione peroxidase, were decreased by DL-alpha-lipoic acid. In conclusion, enhanced oxidative stress is present in rat renal cortex in early diabetes, and is prevented by DL-alpha-lipoic acid.     

Myocardial fibrosis and TGFB expression in hyperhomocysteinemic rats

Hyperhomocysteinemia, characterized by an elevated plasma homocysteine concentration, leads to several clinical manifestations and particularly cardiovascular diseases. Experimental models of hyperhomocysteinemia revealed several tissue injuries including heart fibrosis and ventricular hypertrophy. In order to analyze the molecular mechanisms link to these morphological alterations, a mild hyperhomocysteinemia was induced in rats via a chronic methionine administration. Effects of methionine administration were examined by histological analysis with Sirius red staining, histomorphometric analysis, zymography, and immunoblotting. Hyperhomocysteinemia due to methionine administration produces an interstitial myocardial fibrosis and a ventricular cardiomyocyte hypertrophy, which were associated with increased expression of transforming growth factor-beta1 (TGFβ1), tissue inhibitors of metalloproteinase (TIMP) 2, and JNK activation. However, the matrix metalloproteinase 2 activity was decreased in the hearts of hyperhomocysteinemic rats. Moreover, the TIMP1 protein expression was decreased, and the TIMP1-MMP1 balance was shifted. Remodeling in cardiac tissue observed in rat model of mild hyperhomocysteinemia is associated with a dysregulation in extracellular matrix degradation which results, at least in part, from enhancement of TGFβ1 level.     

ADCK3, an ancestral kinase, is mutated in a form of recessive ataxia associated with coenzyme Q10 deficiency

Muscle coenzyme Q(10) (CoQ(10) or ubiquinone) deficiency has been identified in more than 20 patients with presumed autosomal-recessive ataxia. However, mutations in genes required for CoQ(10) biosynthetic pathway have been identified only in patients with infantile-onset multisystemic diseases or isolated nephropathy. Our SNP-based genome-wide scan in a large consanguineous family revealed a locus for autosomal-recessive ataxia at chromosome 1q41. The causative mutation is a homozygous splice-site mutation in the aarF-domain-containing kinase 3 gene (ADCK3). Five additional mutations in ADCK3 were found in three patients with sporadic ataxia, including one known to have CoQ(10) deficiency in muscle. All of the patients have childhood-onset cerebellar ataxia with slow progression, and three of six have mildly elevated lactate levels. ADCK3 is a mitochondrial protein homologous to the yeast COQ8 and the bacterial UbiB proteins, which are required for CoQ biosynthesis. Three out of four patients tested showed a low endogenous pool of CoQ(10) in their fibroblasts or lymphoblasts, and two out of three patients showed impaired ubiquinone synthesis, strongly suggesting that ADCK3 is also involved in CoQ(10) biosynthesis. The deleterious nature of the three identified missense changes was confirmed by the introduction of them at the corresponding positions of the yeast COQ8 gene. Finally, a phylogenetic analysis shows that ADCK3 belongs to the family of atypical kinases, which includes phosphoinositide and choline kinases, suggesting that ADCK3 plays an indirect regulatory role in ubiquinone biosynthesis possibly as part of a feedback loop that regulates ATP production.     

RNAi depleted Drosophila cell extracts to dissect signaling pathways leading to actin polymerization

Dissection of signal transduction pathways leading to actin polymerization has been performed in cytosolic extracts. In such assays, the implication of an effector molecule is demonstrated by the loss of actin polymerization upon its depletion and the restoration of actin polymerization upon its add-back. Two major limitations in the wide use of this approach have been the availability of immunodepleting antibodies and the functional redundancy for many classes of effector molecules encoded by vertebrate genomes. To circumvent these limitations, we developed extracts derived from S2 Drosophila cells, which are competent for actin polymerization. In this system, depleted extracts are simply obtained from cells cultured with long double stranded RNAs in the medium. We validated the method by showing that beads coated with the C-terminal domain of Wave2 were no longer able to trigger actin polymerization in an extract depleted of the Arp2/3 complex. We also examined the complete set of Drosophila small GTPases of the Rho family for their ability to polymerize actin in such extracts, and found that only dCdc42 was able to induce actin polymerization. Using RNAi depleted extract, we confirmed that dCdc42 triggers actin polymerization in a Wasp dependent manner.     

Aging-Related Systemic Manifestations in COPD Patients and Cigarette Smokers

Rationale

Chronic obstructive pulmonary disease (COPD) is often associated with age-related systemic abnormalities that adversely affect the prognosis. Whether these manifestations are linked to the lung alterations or are independent complications of smoking remains unclear.

Objectives

To look for aging-related systemic manifestations and telomere shortening in COPD patients and smokers with minor lung destruction responsible for a decline in the diffusing capacity for carbon monoxide (DL_CO) corrected for alveolar volume (K_CO).

Methods

Cross-sectional study in 301 individuals (100 with COPD, 100 smokers without COPD, and 101 nonsmokers without COPD).

Measurements and Main Results

Compared to control smokers, patients with COPD had higher aortic pulse-wave velocity (PWV), lower bone mineral density (BMD) and appendicular skeletal muscle mass index (ASMMI), and shorter telomere length (TL). Insulin resistance (HOMA-IR) and glomerular filtration rate (GFR) were similar between control smokers and COPD patients. Smokers did not differ from nonsmokers for any of these parameters. However, smokers with normal spirometry but low K_CO had lower ASMMI values compared to those with normal K_CO. Moreover, female smokers with low K_CO, had lower BMD and shorter TL compared to those with normal K_CO.

Conclusions

Aging-related abnormalities in patients with COPD are also found in smokers with minor lung dysfunction manifesting as a K_CO decrease. Decreased K_CO might be useful, particularly among women, for identifying smokers at high risk for aging-related systemic manifestations and telomere shortening.     

Improvement of fermentative decolorization of olive mill wastewater by Lactobacillus paracasei by cheese whey's addition

The effect of the cheese whey's (CW) addition on the fermentative decolorization of olive mill wastewater (OMW) by Lactobacillus paracasei, with and without pH adjustment by lime, was investigated. Mixtures OMW/CW at different proportions were fermented. The highest colour removal (47%) and total phenolic reduction (22.7%) of OMW were obtained after cofermentation of OMW/CW at proportions of 10/90, respectively. The decrease of pH after cofermentation of the two wastewaters, induced the precipitation of whey proteins with phenolic compounds and, so, improves decolorization. These removal yields reached 64% and 34%, respectively after precipitation by adjustment of pH at 7 with lime at the end of cofermentation. These improvements were correlated to a clarification of wastewaters by precipitation of whey proteins with phenolic compounds. An enhanced decolorization (up to 93%) and a total phenolic reduction (50%) of the mixture were obtained when cofermentation sequentially pH corrected by lime addition was investigated.