Identification of SNP loci and candidate genes genetically controlling norisoprenoids in grape berry based on genome-wide association study.

Obtaining new grapevine varieties with unique aromas has been a long-standing goal of breeders. Norisoprenoids are of particular interest to wine producers and researchers, as these compounds are responsible for the important varietal aromas in wine, characterized by a complex floral and fruity smell, and are likely present in all grape varieties. However, the single-nucleotide polymorphism (SNP) loci and candidate genes genetically controlling the norisoprenoid content in grape berry remain unknown. To this end, in this study, we investigated 13 norisoprenoid traits across two years in an F1 population consisting of 149 individuals from a hybrid of Vitis vinifera L. cv. Muscat Alexandria and V. vinifera L. cv. Christmas Rose. Based on 568,953 SNP markers, genome-wide association analysis revealed that 27 candidate SNP loci belonging to 18 genes were significantly associated with the concentrations of norisoprenoid components in grape berry. Among them, 13 SNPs were confirmed in a grapevine germplasm population comprising 97 varieties, including two non-synonymous mutations SNPs within the VvDXS1 and VvGGPPS genes, respectively in the isoprenoid metabolic pathway. Genotype analysis showed that the grapevine individuals with the heterozygous genotype C/T at chr5:2987350 of VvGGPPS accumulated higher average levels of 6-methyl-5-hepten-2-one and ß-cyclocitral than those with the homozygous genotype C/C. Furthermore, VvGGPPS was highly expressed in individuals with high norisoprenoids concentrations. Transient overexpression of VvGGPPS in the leaves of Vitis quinquangularis and tobacco resulted in an increase in norisoprenoid concentrations. These findings indicate the importance of VvGGPPS in the genetic control of norisoprenoids in grape berries, serving as a potential molecular breeding target for aroma.

Differential Expression of VvLOXA Diversifies C6 Volatile Profiles in Some Vitis vinifera Table Grape Cultivars.

C6 volatiles are synthesized through lipoxygenase-hydroperoxide lyase (LOX-HPL) pathway and these volatiles play important roles in the aromatic quality of grape berries. This study investigated the evolution of both C6 volatiles and the key genes in the LOX-HPL pathway in different table grape cultivars during the berry development period, and further assessed the correlation between the accumulation of C6 volatiles and the expression of these genes in these cultivars. Results showed that hexanal, (E)-2-hexenal, (E)-2-hexen-1-ol and (Z)-3-hexen-1-ol were found to be the dominant C6 volatiles in these ripened grape cultivars under two consecutive vintages, and their flavor notes were incorporated in the overall aroma of these cultivars. The cultivar "Xiangfei" showed the most abundant level of C6 aldehydes and C6 acid, whereas the cultivar "Tamina" and "Moldova" possessed the highest C6 alcohol content. The "Muscat of Alexandria" cultivar was found to contain the highest level of C6 esters. C6 volatiles were grouped into three evolutionary patterns in these cultivars during berry development, and their evolution was consistent with the evolution of the LOX-HPL pathway genes' expression. Pearson's correlation analysis indicated that the LOX-HPL-pathway-related genes were correlated to the accumulation of C6 volatiles in these cultivars, and VvLOXA appeared to be an important gene that regulated the synthesis of all C6 volatiles.

Application of CRT-D in a Marfan syndrome patient with chronic heart failure accompanied by ventricular tachycardia and ventricular fibrillation.

Marfan syndrome is a systemic connective tissue disease that could affect the cardiovascular system and eventually lead to heart enlargement and heart failure with high mortality, mainly due to progressive heart failure and/or sudden cardiac death caused by malignant arrhythmia. Here we report that a patient received a cardiac resynchronization therapy-defibrillator (CRT-D) with a pre-monitor function for heart failure and experienced obvious improvements in his cardiac function. Postoperative follow-up showed that the patient had reduced morbidity and hospitalization for heart failure, and also experienced improved quality of life.

Efficacy and safety of nitroglycerin for preventing venous spasm during contrast-guided axillary vein puncture for pacemaker or defibrillator leads implantation.

AIMS: We investigated the efficacy and safety of nitroglycerin for preventing venous spasm during contrast-guided axillary vein puncture for pacemaker or defibrillator leads implantation. METHODS AND RESULTS: A total of 40 consecutive patients referred for contrast-guided axillary vein puncture for pacemaker or defibrillator implantations were included in the study. Patients were randomly assigned to control group and nitroglycerin group. Patients in the nitroglycerin group were given 200 µg (2 mL) nitroglycerin via ipsilateral peripheral vein about 3 min before puncture. The degree of venous spasm was evaluated by the reduction in lumen calibre of the axillary vein after puncture. Mild venous spasm and severe venous spasm were defined as a reduction in lumen calibre of 50-90% and ≥ 90%, respectively. The mean degree of venous spasm of axillary vein was lower in the nitroglycerin group than in the control group (23.0 ± 22.3 vs. 45.5 ± 33.6%, P = 0.018). The incidence of mild or severe venous spasm was lower in the nitroglycerin group than in the control group (3/20 vs. 11/20, P = 0.019). In the nitroglycerin group, the systolic blood pressure had a significant decrease after puncture (129.5 ± 23.7 vs. 143.0 ± 24.1 mmHg, P = 0.003). There was no hypotension and other adverse reaction of nitroglycerin in the nitroglycerin group. CONCLUSION: Intravenous nitroglycerin is effective and safe for preventing venous spasm during contrast-guided axillary vein puncture for pacemaker or defibrillator leads implantation.

Venous spasm during contrast-guided axillary vein puncture for pacemaker or defibrillator lead implantation.

AIMS: Contrast-guided axillary vein puncture is commonly used for pacemaker or defibrillator lead implantation. Venous spasm during contrast-guided axillary vein puncture has been reported rarely. We investigated the incidence of venous spasm during contrast-guided axillary vein puncture and the impact of venous spasm on axillary vein puncture. METHODS AND RESULTS: Seventy-four consecutive patients referred for contrast-guided axillary vein puncture for pacemaker or defibrillator implantation were included in the study. The puncture was guided by fluoroscopy and the venogram of axillary vein. After the success of puncture, the venography was taken again. When the puncture could not be successful up to 3 min, the second venography was taken and the puncture was guided by the second venogram. The degree of venous spasm was evaluated by the reduction in the lumen calibre of the axillary vein after puncture. Mild venous spasm and severe venous spasm were defined as a reduction in the lumen calibre of 50-90 and ≥ 90%, respectively. The success rate of axillary vein puncture within 3 min was 87.8%, and the total success rate was 95.9%. Mild venous spasm occurred in 22 patients (29.7%) and severe venous spasm occurred in 6 patients (8.1%). Severe venous spasm occurred in all the three patients with a failed puncture. Severe venous spasm was independently negatively correlated with both success rate within 3 min and total success rate. CONCLUSION: Venous spasm is not a rare phenomenon during the contrast-guided puncture of axillary vein and it has an important negative impact on the puncture.

Association between transforming growth factor beta1 polymorphisms and left ventricle hypertrophy in essential hypertensive subjects.

Left ventricular hypertrophy (LVH) increases the risk of cardiovascular morbid events in hypertension. TGF-beta1 is involved in pathologic states such as cardiac hypertrophy and cardiac fibrosis; we thus postulate that the TGF-beta1 polymorphism is related to LVH in hypertensives. Six hundred and eighty essential hypertensive patients were recruited. Biochemical variables and clinical data were obtained and the determination of LVH was performed by echocardiography. According to the presence of LVH, all subjects were divided into the LVH+ and LVH- group. DNA was obtained, and two coding region polymorphisms of the TGF-beta1 gene (+869 Leu-->Proat codon 10 and +915 ARG-->Pro at codon 25) were analyzed by the polymerase chain reaction. The product was cleaved with the restriction endonucleases. For the polymorphisms of the +869 Leu-->Pro at codon 10, there was no marked difference in the distributions of genotypes and the allele frequencies between the LVH+ and LVH- subjects. For +915 Arg-->Pro at codon 25, a significant difference in the distributions of genotypes of TGF-beta1 was observed. The left ventricular mass index (LVMI) in Arg-Pro genotype carriers was significantly higher than those in the Arg-Arg and Pro-Pro carriers. Multivariate analysis showed that the Arg-Pro genotype was an independent risk factor for LVH (OR 3.23, 95% CI [1.48-5.63, P = 0.002]). The codon 10 genotypes did not show a significant association to LVH. Our data revealed a genetic association of TGF-beta1+915 Arg-->Pro at codon 25 polymorphism with LVH in a Chinese hypertensive population.

Lishizhenia tianjinensis sp. nov., isolated from coastal seawater.

A Gram-negative, non-carbohydrate-utilizing, non-flagellated, motile, flexible, long rod-shaped, orange-pigmented bacterium, strain H6(T), was isolated from coastal seawater of Tianjin City, China, and its taxonomic position was investigated by using a polyphasic approach. Strain H6(T) grew optimally at 30 degrees C, in the presence of 2.0 % (w/v) NaCl and at pH 7.6. Menaquinone-6 (MK-6) was the sole respiratory quinone and the major fatty acids were iso-C(15 : 0) (36.5 % of the total), iso-C(15 : 1) (27.3 %) and iso-C(17 : 0) 3-OH (10.8 %). The DNA G+C content of strain H6(T) was 34.6 mol%. Phylogenetic analysis based on 16S rRNA gene sequences demonstrated that strain H6(T) was related most closely to Lishizhenia caseinilytica JCM 13821(T) (97.8 % similarity). DNA-DNA relatedness between the two strains was 47.4 %. Strain H6(T) could be further differentiated from L. caseinilytica JCM 13821(T) based on activity of several enzymes, hydrolysis of casein and Tweens 40, 60 and 80 and production of H(2)S. On the basis of phenotypic, chemotaxonomic, genomic and phylogenetic data, strain H6(T) is considered to represent a novel species of the genus Lishizhenia, for which the name Lishizhenia tianjinensis sp. nov. is proposed. The type strain is H6(T) (=CGMCC 1.7005(T) =JCM 15141(T)).

Zhangella mobilis gen. nov., sp. nov., a new member of the family Hyphomicrobiaceae isolated from coastal seawater.

A Gram-negative, pale-yellow, non-spore-forming bacterium, motile with single polar flagella, strain E6T, was isolated from coastal seawater collected from Tianjin, China, and its taxonomic position was investigated using a polyphasic approach. Strain E6T requires NaCl for growth and grows optimally at pH 8.2 and 36 degrees C and in the presence of 2.0% (w/v) NaCl. It is positive for catalase and oxidase, and reduces nitrate to nitrite. The major fatty acids (>10%) are C18:1omega7c (64.71%) and 11-methyl C18:1omega7c (12.38%), the ubiquinone system is Q-10 and the DNA G+C content is 53.1 mol%. Phylogenetic analyses based on 16S rRNA gene sequences demonstrated that strain E6T represented a new lineage in the Alphaproteobacteria and is related to genera Cucumibacter, Devosia, Ochrobactrum and Ahrensia. Strain E6T shows highest 16S rRNA gene sequence similarity (93.3%) to Cucumibacter marinus CL-GR60T and less than 92.2% similarity to other relatives. It can be differentiated from its closest phylogenetic neighbours on the basis of several phenotypic features, including nitrate reduction, assimilation of D-glucose, L-arabinose, D-mannose, mannitol and maltose and major fatty acid composition. A polyphasic analysis supported the conclusion that strain E6T represents a novel genus and species of the family Hyphomicrobiaceae, for which the name Zhangella mobilis gen. nov., sp. nov. is proposed. The type strain of Zhangella mobilis is E6T (=CGMCC 1.7002T=JCM 15144T).

Protective role of granulocyte colony-stimulating factor against adriamycin induced cardiac, renal and hepatic toxicities.

Adriamycin (ADR) causes dose-dependant toxicities in heart, liver and kidneys via inducing the peroxidative alterations in organ tissues. Recent studies showed that the granulocyte colony-stimulating factor (G-CSF) exerts beneficial effects on heart, liver and kidney injuries induced by different pathological conditions. We hypothesize that G-CSF have a protective effect on ADR induced cardiac, renal and hepatic toxicities by inhibiting the peroxidative alterations in organ tissues. Wistar rats were randomly divided into control, ADR, ADR+phosphate buffered saline (PBS) and ADR+G-CSF group (n=16 in each group). ADR was administered intraperitoneally every other day at the dose of 2.5 microg/kg each time per rat (total six times of injection during 2 weeks). Rats in the ADR+G-CSF group were injected subcutaneously with G-CSF at the dose of 50 microg/(kg day) (for 8 consecutive days). After 8 weeks, the serum and urine biochemistry variables were determined. The malondialdehyde (MDA) level and the glutathione (GSH) content in the heart, the liver and the kidney tissues were measured. ADR caused significant cardiac, renal and hepatic toxicities indicated by the serum and urine biochemistry variables. The tissue MDA level in the heart, kidney and liver in rats treated with ADR were markedly elevated, while the GSH content in these tissues were significantly reduced. G-CSF administration palliated the cardiac, renal and hepatic toxicities. Notably, G-CSF induced significant reduction of MDA level and increase of GSH content in the heart, kidney and liver tissues. This study suggests that G-CSF play an overall protective effect on ADR-induced toxicities in heart, liver and kidneys and the inhibition of tissue peroxidative alterations might contribute to this beneficial effect.

Glaciecola lipolytica sp. nov., isolated from seawater near Tianjin city, China.

A Gram-negative, non-spore-forming, rod-shaped bacterium, designated strain E3T, which was motile with a polar flagellum, was isolated from coastal surface seawater near Tianjin city, China. The taxonomy of strain E3T was studied by phenotypic, chemotaxonomic and phylogenetic methods. Strain E3T forms non-pigmented colonies with a butyrous-mucoid consistency on marine 2216 agar. It grows optimally at 30 degrees C, in the presence of 2.0 % (w/v) NaCl and at pH 7.6. Strain E3T is positive for catalase and oxidase and hydrolysis of lipid and reduces nitrate to N2 gas. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain E3T belongs to the Gammaproteobacteria and is grouped in the genus Glaciecola, with highest sequence similarity to the type strains of Glaciecola chathamensis (94.6 %), G. agarilytica (94.4 %) and G. mesophila (94.4 %). Strain E3T contained summed feature 3 (C(16 : 1)omega7c and/or iso-C(15 : 0) 2-OH, 34.5 %), C(18 : 1)omega7c (17.3 %) and C(16 : 0) (17 %) as the major fatty acids and phosphatidylethanolamine and phosphatidylglycerol as the main phospholipids. The DNA G+C content was 40.8 mol%. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain E3T is considered to represent a novel species of the genus Glaciecola, for which the name Glaciecola lipolytica sp. nov. is proposed. The type strain is E3T (=CGMCC 1.7001T =JCM 15139T).

[Puerarin suppresses the proliferation of vscular smooth muscule cells and c-fos and bcl-2 protein expression].

OBJECTIVE: To observe the role of puerarin on the proliferation of vascular smooth muscle cells(VSMC) induced by thrombin (T) and the effect of puerarin on the c-fos and bcl-2 protein expression. METHOD: Cell number and cell cycle analysis using flow cytometry were adopted as two different indicators of effects on proliferation of VSMC. Western blot was used to indicate the changes of c-fos and bcl-2 protein after 24 h of treatment of T and puerarin. RESULT: 1.5 x 10(-5) - 1.5 x 10(-3) mol x L(-1) puerarin could significantly suppress this stimulation of VSMC proliferation and DNA synthesis induced by T. Western blot demonstrated that after 24 hour of treatment with T and puerarin, T could significantly increase c-fos and bcl-2 protein and 1.5 x 10(-5) - 1.5 x 10(-3) mol x L(-1) puerain could significantly suppress this increase. CONCLUSION: puerarin can suppress the proliferation and DNA synthesis of VSMC promoted by T. This inhibitory effects of puerarin are closely related with the suppression of c-fos and bcl-2 protein.

Effects of 4-hydroxy-2-nonenal on cultured human aortic endothelial cells and myocardial cell.

To study the effects of 4-hydroxy-2-nonenal (HNE) on cultured human aortic endothelial cells and myocardial cells so as to explore the mechanism of the pathogenesis of atherosclerosis. In situ cell death technique, quantitative DNA damage detection and immunohistochemistry were used to identify the cell apoptosis and DNA damage in cultured human aortic endothelial cells and myocardial cells. Tail moment was 32.80+/-1.12, 44.30+/-0.99 and 74.6+/-0.97 when HAOEC were treated with 5 muM, 10muM and 15 muM of HNE for 10 hours, which were of statistical significance when compared with the normal group (6.0+/-0.67, P < 0.001 respectively), But when HAOEC was treated with 1 muM of HNE, the tail moment was 11.3+/-0.9, which was of no statistical difference compared with the untreated group(P>0.05). When human aortic endothelial cells (HAOEC) were treated with 5 muM, 10muM and 15 muM of HNE for 10 hours, the percent of nonviable cells were 5.70+/-0.55, 25.96+/-2.02 and 50.80+/-3.40 (P<0.001 respectively when compared with the normal group with the percent of 0.27+/-0.13). But when HAOEC was treated with 1 muM of HNE for 10 hours, the percent of nonviable cells was 2.5+/-0.22, and no difference was observed when compared with the untreated group (P>0.05). When cultured human myocardial cells were treated with 5 muM of HNE for 10 hours, TUNEL staining showed a greater number of apoptotic cells in HNE-treated human myocardial cells. No TUNEL-positive cells were observed in untreated group. When HAOEC was treated with 5 muM of HNE for 10 hours, immunocytochemical labeling with polyclonal antibody to HNE-modified proteins revealed specific cytoplasmic staining in cells incubated with HNE, whereas staining was absent in control cells incubated with vehicle. But 1 muM of HNE treatment didn't present positive stainings. Higher concentrations of HNE (10 muM and 15 muM) showed much stronger positive stainings. HNE induces DNA damage and cell apoptosis of cultured aortic endothelial cells and myocardial cells. The DNA damage and apoptosis levels are proportional to the HNE concentrations.